Information on EC 3.1.1.74 - cutinase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
3.1.1.74
-
RECOMMENDED NAME
GeneOntology No.
cutinase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cutin + H2O = cutin monomers
show the reaction diagram
Cutin, a polymeric structural component of plant cuticles, is a hydroxy fatty acid polymer, usually C16 or C18 and that contains one to three hydroxyl groups. The enzyme from several fungal sources also hydrolyses the p-nitrophenyl esters of hexadecanoic acid. It is however inactive towards several esters that are substrates for non-specific esterases.
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
carboxylic ester hydrolysis
-
-
-
-
carboxylic ester hydrolysis
A8QPD8, -
-
hydrolysis
-
-
SYSTEMATIC NAME
IUBMB Comments
cutin hydrolase
Cutin, a polymeric structural component of plant cuticles, is a polymer of hydroxy fatty acids that are usually C16 or C18 and contain up to three hydroxy groups. The enzyme from several fungal sources also hydrolyses the p-nitrophenyl esters of hexadecanoic acid. It is however inactive towards several esters that are substrates for non-specific esterases.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
CcCUT1
B9U443
-
CDEF1
-
-
CDEF1
Q9SZW7
-
Cut1
P00590
-
Cut1
Thermobifida cellulosilytica DSM44535
-
-
-
Cut1
Thermobifida fusca DSM44342
-
-
-
Cut2
Thermobifida cellulosilytica DSM44535
-
-
-
cuticle destructing factor 1
-
-
cuticle destructing factor 1
Q9SZW7
-
cutin esterase
-
-
-
-
cutinase
P41744
-
cutinase
-
-
cutinase
Q9SZW7
-
cutinase
B9U443
-
cutinase
-
-
cutinase
P00590
-
cutinase
-
-
cutinase
P11373
-
cutinase
-
-
cutinase
A8QPD8
-
cutinase
Magnaporthe grisea, Neurospora crassa, no activity in Saccharomycotina
-
-
cutinase
-
-
cutinase
-
-
cutinase-1
P00590
-
cutinolytic polyesterase
B9U443
-
Tfu_0883
-
-
Thcut1
A8QPD8
-
THCUT1 protein
A8QPD8
-
fungal cutinase
-
-
additional information
-
the cutinase is an esterase that belongs to the alpha/beta hydrolases family
CAS REGISTRY NUMBER
COMMENTARY
51377-41-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Botryotinia fuckeliana Pser.: Fr.
Pser.: Fr.
-
-
Manually annotated by BRENDA team
Coprinopsis cinerea C29-2
-
-
-
Manually annotated by BRENDA team
strain 15; strain 20; strain 211; strain 35; strain CECT
-
-
Manually annotated by BRENDA team
Emericella nidulans 15
strain 15
-
-
Manually annotated by BRENDA team
Emericella nidulans 20
strain 20
-
-
Manually annotated by BRENDA team
Emericella nidulans 211
strain 211
-
-
Manually annotated by BRENDA team
Emericella nidulans 35
strain 35
-
-
Manually annotated by BRENDA team
Emericella nidulans CECT
strain CECT
-
-
Manually annotated by BRENDA team
gladioli; lini; lycopersici; lycopersici 2
-
-
Manually annotated by BRENDA team
sp. pisi
-
-
Manually annotated by BRENDA team
Fusarium roseum
culmorum
-
-
Manually annotated by BRENDA team
cutinase I and cutinase II; pisi
-
-
Manually annotated by BRENDA team
pisi
SwissProt
Manually annotated by BRENDA team
pisi 4
SwissProt
Manually annotated by BRENDA team
recombinantly expressed in Escherichia coli
SwissProt
Manually annotated by BRENDA team
recombinantly expressed in in Escherichia coli WK-6
-
-
Manually annotated by BRENDA team
native enzyme and mutant myHiC with increased activity and decreased surfactanct sensitivity
-
-
Manually annotated by BRENDA team
recombinantly expressed in Aspergillus oryzae
-
-
Manually annotated by BRENDA team
(Wint.) Honey
-
-
Manually annotated by BRENDA team
enzyme precursor
SwissProt
Manually annotated by BRENDA team
expression in Escherichia coli
-
-
Manually annotated by BRENDA team
expression in Saccharomyces cerevisiae
-
-
Manually annotated by BRENDA team
no activity in Saccharomycotina
-
-
-
Manually annotated by BRENDA team
Thc_Cut1 and Thc_Cut2
-
-
Manually annotated by BRENDA team
Thermobifida cellulosilytica DSM44535
Thc_Cut1 and Thc_Cut2
-
-
Manually annotated by BRENDA team
ATCC 27730
-
-
Manually annotated by BRENDA team
Thf42_Cut1
-
-
Manually annotated by BRENDA team
two isozymes Tfu 0882 and Tfu 0883
-
-
Manually annotated by BRENDA team
Thermobifida fusca DSM44342
Thf42_Cut1
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
cutinases are hydrolytic enzymes that share properties of lipases and esterases, they are active regardless of the presence of an interface
physiological function
-
cutinases are hydrolytic enzymes that share properties of lipases and esterases, and also display the unique characteristic of being active regardless of the presence of an interface
physiological function
-
cutinases are hydrolytic enzymes that share properties of lipases and esterases, they are active regardless of the presence of an interface
physiological function
-
the organism contains 11 cutinases, despite the 3-4% repetitive DNA content and the repeat induced point mutation-based elimination of transposable elements
physiological function
-
contains four cutinase genes, which may result from its low repetitive content and mild form of repeat induced point mutation
physiological function
-
the organism contains 12 cutinases. High number of cutinases likely reflects its needs during post-invasion necrotrophic growth and overwintering as saprotrophic mycelia, and its ability to infect many different monocotyledonous genera asymptomatically
physiological function
-
the organism contains 17 cutinases. Preservation of a large number of diverse cutinases within the genome may provide the fungus with a great selective advantage to breach multiple, diverse grass cuticles, or may reflect its requirements to degrade different plant debris while overwintering as a soil saprotroph
physiological function
-
contains three cutinases, which show less than 80% sequence identity, indicating that they are duplicated and diverged before the emergence of the active repeat induced point mutation defence mechanism, and have been retained in the genome by virtue of their varying regulatory or functional diversity
physiological function
-
ectopic expression of CDEF1 driven by the 35S promoter causes fusion of organs, including leaves, stems and flowers, and increased surface permeability
physiological function
Q9SZW7
CDEF1 is a plant cutinase, recombinant CDEF1 protein has esterase activity. Ectopic expression of CDEF1 driven by the 35S promoter causes fusion of organs, including leaves, stems and flowers, and increased surface permeability. CDEF1 is involved in the penetration of the stigma by pollen tubes. CDEF1 degrades cell wall components to facilitate the emergence of the lateral roots
additional information
-
biophysical parameters of cutinase as a function of pH, overview
additional information
-
biophysical parameters of cutinase as a function of pH, overview
additional information
-
biophysical parameters of cutinase as a function of pH, overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,4-butanediol + adipic acid
?
show the reaction diagram
-
polycondensation reaction, degree of polymerization is 13
-
-
?
1,4-cyclohexanedimethanol + adipic acid
?
show the reaction diagram
-
polycondensation reaction, degree of polymerization is 16
-
-
?
1,4-cyclohexanedimethanol + sebacic acid
?
show the reaction diagram
-
polycondensation reaction, degree of polymerization is 61
-
-
?
1,4-cyclohexanedimethanol + suberic acid
?
show the reaction diagram
-
polycondensation reaction, degree of polymerization is 18
-
-
?
1,4-cyclohexanedimethanol + succinic acid
?
show the reaction diagram
-
polycondensation reaction, degree of polymerization is 4
-
-
?
1,8-octanediol + adipic acid
?
show the reaction diagram
-
polycondensation reaction, degree of polymerization is 47
-
-
?
2-hydroxyethyl benzoate + H2O
ethane-1,2-diol + benzoate
show the reaction diagram
-
is hydrolyzed after 24 h of incubation of bisbenzoyloxyethyl terephthalate
-
-
?
4-nitrophenyl acetate
4-nitrophenol + acetate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl acetate + H2O
4-nitrophenol + acetate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl acetate + H2O
4-nitrophenol + acetate
show the reaction diagram
A8QPD8, -
-
-
-
?
4-nitrophenyl acetate + H2O
4-nitrophenol + acetate
show the reaction diagram
Thermobifida cellulosilytica, Thermobifida fusca DSM44342, Thermobifida cellulosilytica DSM44535
-
-
-
-
?
4-nitrophenyl butanoate + H2O
4-nitrophenol + butanoate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl butyrate
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
-
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
-
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
-
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
-
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
A8QPD8, -
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
Q9SZW7
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-, P41744
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
A6N6J6
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
A6N6J6
-
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
Thermobifida fusca DSM44342, Thermobifida cellulosilytica DSM44535
-
-
-
-
?
4-nitrophenyl dodecanoate + H2O
4-nitrophenol + dodecanoate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl dodecanoate + H2O
4-nitrophenol + dodecanoate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl hexanoate
4-nitrophenol + hexanoate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl octanoate + H2O
4-nitrophenol + octanoate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl octanoate + H2O
4-nitrophenol + octanoate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl palmitate + H2O
4-nitrophenol + palmitate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl palmitate + H2O
4-nitrophenol + palmitate
show the reaction diagram
A8QPD8, -
-
-
-
?
4-nitrophenyl palmitate + H2O
4-nitrophenol + palmitate
show the reaction diagram
-, P41744
-
-
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl valerate
4-nitrophenol + pentanoate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl valerate + H2O
4-nitrophenol + pentanoate
show the reaction diagram
A8QPD8, -
-
-
-
?
4-nitrophenylbutyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
4-nitrophenylpalmitate + H2O
4-nitrophenol + palmitate
show the reaction diagram
Coprinopsis cinerea, Coprinopsis cinerea C29-2
-
-
-
-
?
beta-butyrolactone
?
show the reaction diagram
-
ring-opening polymerizations
-
-
?
bis(2-hydroxyethyl)terephthalate + H2O
?
show the reaction diagram
-
fast hydrolysis after treatment for 30 min
-
-
?
bis(2-hydroxyethyl)terephthalate + H2O
?
show the reaction diagram
-
hydrolysis after treatment for 5 h
-
-
?
cutin + H2O
cutin monomers
show the reaction diagram
B9U443, -
-
-
-
?
cutin + H2O
cutin monomers
show the reaction diagram
A8QPD8, -
-
-
-
?
cutin + H2O
cutin monomers
show the reaction diagram
-
-
main products: hexadecaoic acid, octadecaoic acid, and 10,16-dihydroxyhexadecaoic acid
-
?
cyclohexyl hexadecanoate + H2O
hexadecanoic acid + cyclohexanol
show the reaction diagram
-
-
-
-
?
delta-valerolactone
?
show the reaction diagram
-
ring-opening polymerizations
-
-
?
dihexyl phthalate + H2O
1,3-isobenzofurandione + ?
show the reaction diagram
-
-
-
-
?
dihexylphthalate + H2O
?
show the reaction diagram
-
degradation by cutinase is nearly 70% after 4 h, while 85% of the initial amount remains intact after 72 h of incubation with Candida cylindracea esterase. Products of cutinase-catalyzed hydrolysis are less toxic than those employing Candida cylindracea esterase
-
-
?
dipentyl phthalate + H2O
?
show the reaction diagram
-
degradation rate of fungal cutinase is high, i.e., almost 60% of the initial dipentyl phthalate is decomposed within 2.5 hours, and nearly 40% of the degraded dipentyl phthalate disappears within the initial 15 min
-
-
?
dipropyl phthalate + H2O
1,3-isobenzofurandione + propanol
show the reaction diagram
-
-
-
-
?
epsilon-caprolactone
?
show the reaction diagram
-
ring-opening polymerizations
-
-
?
ethyl butyrate + H2O
butyric acid + ethanol
show the reaction diagram
-, P41744
-
-
-
?
ethyl caprylate + H2O
caprylic acid + ethanol
show the reaction diagram
-, P41744
-
-
-
?
hexadecyl hexadecanoate + H2O
hexadecanoic acid + hexadecanol
show the reaction diagram
-
weak activity
-
-
?
malathion + H2O
?
show the reaction diagram
-
-
-
-
?
malathion + H2O
malathion monoacid + malathion diacid + ethanol
show the reaction diagram
-
60% of initial 500 mg/l malathion are degraded within 0.5 h
diacid is the major degradation product
-
?
methyl acetate + H2O
acetic acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
methyl butyrate + H2O
butyric acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
methyl caproate + H2O
caproic acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
methyl caprylate + H2O
caprylic acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
methyl decanoate + H2O
decanoic acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
methyl hexadecanoate + H2O
hexadecanoic acid + methanol
show the reaction diagram
-
-
-
-
?
methyl laurate + H2O
lauric acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
methyl myristate + H2O
myristic acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
n-butyl benzyl phthalate + H2O
1,3-isobenzofurandione + n-butanol + benzyl alcohol
show the reaction diagram
-
-
major product, less than 5% of byproducts such as dimethyl phthalate, butyl methyl phthalate
-
?
omega-pentadecalactone
?
show the reaction diagram
-
ring-opening polymerizations
-
-
?
p-nitrophenyl acetate + H2O
p-nitrophenol + acetate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl acetate + H2O
p-nitrophenol + acetate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl acetate + H2O
p-nitrophenol + acetate
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl butyrate + H2O
p-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl butyrate + H2O
p-nitrophenol + butyrate
show the reaction diagram
P00590
-
-
-
?
p-nitrophenyl butyrate + H2O
p-nitrophenol + butyrate
show the reaction diagram
-
molecular modelling allows the synthesis of a solid-phase combinatorial library of triazine-based synthetic affinity compounds that is assessed for binding cutinase with high affinity while preserving enzyme functionality. Detection of binding ligands, in which immobilized cutinase retains 3060% of its enzymatic activity as compared to free enzyme
-
-
?
p-nitrophenyl butyrate + H2O
p-nitrophenol + butyrate
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl butyrate + H2O
phenol + butyrate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl caprate + H2O
p-nitrophenol + ?
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl caproate + H2O
p-nitrophenol + ?
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl hexanoate + H2O
p-nitrophenol + hexanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl hexanoate + H2O
p-nitrophenol + hexanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl laurate + H2O
p-nitrophenol + ?
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl myristate + H2O
p-nitrophenol + ?
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl palmitate + H2O
p-nitrophenol + ?
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl propionate + H2O
p-nitrophenol + ?
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenyl stearate + H2O
p-nitrophenol + ?
show the reaction diagram
B9U443, -
a lower concentration of p-nitrophenyl stearate (2.5 mM) is used due to its lower solubility
-
-
?
p-nitrophenyl valerate + H2O
p-nitrophenol + pentanoate
show the reaction diagram
A8QPD8, -
-
-
-
-
p-nitrophenyl valerate + H2O
p-nitrophenol + pentanoate
show the reaction diagram
B9U443, -
concentration of substrate dispersion is 5 mM
-
-
?
p-nitrophenylbutanoate + H2O
p-nitrophenol + butanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenylbutanoate + H2O
p-nitrophenol + butanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenylbutanoate + H2O
p-nitrophenol + butanoate
show the reaction diagram
Fusarium roseum
-
-
-
-
?
p-nitrophenylbutanoate + H2O
p-nitrophenol + butanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenylbutanoate + H2O
p-nitrophenol + butanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenylbutanoate + H2O
p-nitrophenol + butanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenylbutanoate + H2O
p-nitrophenol + butanoate
show the reaction diagram
Q8TGB8
-
-
-
?
p-nitrophenyldecanoate + H2O
p-nitrophenol + decanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenyldecanoate + H2O
p-nitrophenol + decanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenylhexadecanoate + H2O
p-nitrophenol + hexadecanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenyltetradecanoate + H2O
p-nitrophenol + tetradecanoate
show the reaction diagram
-
-
-
-
?
poly(ethylene terephthalate) + H2O
mono-(2-hydroxyethyl) terephthalate + terephthalic acid
show the reaction diagram
-
-
no formation of bis(2-hydroxyethyl) terephthalate, terephthalic acid is the major hydrolysis product for Thc_Cut1, whereas for Thc_Cut2, mono-(2-hydroxyethyl) terephthalate is the most abundant product
-
?
poly(ethylene terephthalate) + H2O
mono-(2-hydroxyethyl) terephthalate + terephthalic acid
show the reaction diagram
Thermobifida fusca, Thermobifida fusca DSM44342
-
-
no formation of bis(2-hydroxyethyl) terephthalate. Terephthalic acid is the major hydrolysis product for Thf42_Cut1
-
?
poly(ethylene terephthalate) + H2O
mono-(2-hydroxyethyl) terephthalate + terephthalic acid
show the reaction diagram
Thermobifida cellulosilytica DSM44535
-
-
no formation of bis(2-hydroxyethyl) terephthalate, terephthalic acid is the major hydrolysis product for Thc_Cut1, whereas for Thc_Cut2, mono-(2-hydroxyethyl) terephthalate is the most abundant product
-
?
polyamide + H2O
?
show the reaction diagram
-
-
-
-
?
polyethylene terephthalate + H2O
terephthalate + ?
show the reaction diagram
-
-
-
-
?
suberin + H2O
?
show the reaction diagram
B9U443, -
9,10-epoxy-18-hydroxy 5 octadecanoic acid
-
-
?
tributyrin + H2O
?
show the reaction diagram
-
-
-
-
?
tributyrin + H2O
?
show the reaction diagram
-
-
-
-
?
tributyrin + H2O
?
show the reaction diagram
-, P41744
-
-
-
?
tricaprylin + H2O
?
show the reaction diagram
-
-
-
-
?
tricaprylin + H2O
?
show the reaction diagram
-, P41744
-
-
-
?
triglyceride + H2O
?
show the reaction diagram
-
triglycerides in which one of the primary acyl ester functions has been replaced by an alkyl grpup and the secondary acyl ester bond has been replaced by an acyl amino bond. The activity is very sensitive to the length and distribution of the acyl chains, the highest activity is found when the chains at position 1 and 3 contain three or four carbon atoms
-
-
?
triolein + H2O
?
show the reaction diagram
-
-
-
-
?
triolein + H2O
?
show the reaction diagram
-, P41744
-
-
-
?
tripalmitin + H2O
?
show the reaction diagram
-, P41744
-
-
-
?
tristearin + H2O
?
show the reaction diagram
-, P41744
-
-
-
?
bisbenzoyloxyethyl terephthalate + H2O
terephthalic acid + mono(2-hydroxyethyl) terephthalate + bis(2-hydroxyethyl)terephthalate + benzoic acid + 2-hydroxyethyl benzoate
show the reaction diagram
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
P00590
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
Fusarium roseum
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
-
-
-
-
?
cutin + H2O
additional information
-
-
-
dihydroxyhexadecanoic acid, cutin monomer + cutin oligomers
?
cutin + H2O
additional information
-
-
apple cutin
-
-
?
cutin + H2O
additional information
-
Emericella nidulans 211, Emericella nidulans 15, Emericella nidulans 35, Emericella nidulans CECT, Emericella nidulans 20
-
-
-
-
?
cutin + H2O
additional information
-
Botryotinia fuckeliana Pser.: Fr.
-
-
-
-
?
methyl propionate + H2O
propionic acid + methanol
show the reaction diagram
-, P41744
-
-
-
?
additional information
?
-
-
-
-
-
-
additional information
?
-
-
does not act on tripalmitoyl glycerol or trioleoyl glycerol
-
-
-
additional information
?
-
Fusarium roseum
-
no hydrolysis of p-nitrophenyl palmitate
-
-
-
additional information
?
-
-
catalytic triad: S120, H188, D175. Presence of a preformed oxyanion hole
-
-
-
additional information
?
-
-
constitutive enzyme
-
-
-
additional information
?
-
-
induced by cutin
-
-
-
additional information
?
-
-
Cutinase is known for its hydrolytic activity for a variety of esters ranging from soluble p-nitrophenyl esters to insoluble long-chain triglycerides. The hydrolytic activity of cutinase, especially on p-nitrophenyl esters of fatty acids, is extremely sensitive to fatty acid chain length.
-
-
-
additional information
?
-
B9U443, -
CcCUT1 has higher activity on shorter (C2-C10) 12 fatty acid esters of p-nitrophenol than on longer ones and it also exhibited lipase activity. Microscopical analyses and determination of released hydrolysis products showed that the enzyme is able to depolymerize apple cutin and birch outer bark suberin
-
-
-
additional information
?
-
-
during its catalytic cycle, cutinase undergoes a significant conformational rearrangement converting the loop bearing the histidine from an inactive conformation, in which the histidine of the triad is solvent exposed, to an active conformation, in which the triad assumes a classic configuration. Major difference between the structures is in the position of the loop connecting beta5 and alpha5 (Gly196Phe205 in Glomerella cingulata cutinase and Gly180Leu189 in Fusarium solani cutinase). Consequence of the repositioning of the loop is that the active-site regions of the enzymes differ substantially in the location of the putative catalytic histidine (His188 of Fusarium solani cutinase and His204 of Glomerella cingulata cutinase)
-
-
-
additional information
?
-
-
four cysteine residues pivotal to the formation of the two disulphide bridges and a highly conserved cut-1 motif (GYSQG) surrounding a cutinase active serine, but a less precise cut-2 motif, DxVCxG(ST)-(LIVMF)(3)-x(3)H, which carries the aspartate and histidine residues of the active site
-
-
-
additional information
?
-
-
four cysteine residues pivotal to the formation of the two disulphide bridges and a highly conserved cut-1 motif (GYSQG) surrounding a cutinase active serine, but a less precise cut-2 motif, DxVCxG(ST)-(LIVMF)(3)-x(3)H, which carries the aspartate and histidine residues of the active site. Two exceptions: one cutinase gene is truncated at the 3' end immediately after the cut-1 motif owing to a gap in the genomic sequence, and one cutinase gene, which is truncated at the 3' end shortly before the cut-2 motif because of a repetitive sequence, making further prediction impossible
-
-
-
additional information
?
-
-
major difference between the structures is in the position of the loop connecting beta5 and alpha5 (Gly196Phe205 in Glomerella cingulata cutinase and Gly180Leu189 in Fusarium solani cutinase). Consequence of the repositioning of the loop is that the active-site regions of the enzymes differ substantially in the location of the putative catalytic histidine (His188 of Fusarium solani cutinase and His204 of Glomerella cingulata cutinase)
-
-
-
additional information
?
-
-
shows promising activity in polymerization reactions
-
-
-
additional information
?
-
-
cutinase is an esterase, whose active site, located at the middle of a sharp turn between beta-strand and alpha-helix, is composed by the triad Ser120, Asp175 and His188
-
-
-
additional information
?
-
-
the enzyme exhibits a broad substrate specificity against plant cutin, synthetic polyesters, insoluble triglycerides, and soluble esters
-
-
-
additional information
?
-
-
cutinase catalyzes esterification of caproic acid in an organic solvent system, alcohol, acid and n-decane are mixed thoroughly in iso-octane before the addition of the lyophilized enzyme, overview. The main kinetic characteristics observed in esterification reaction follow an ordered Ping-Pong Bi-Bi mechanism
-
-
-
additional information
?
-
-
substrate binding, modelling and docking study, overview
-
-
-
additional information
?
-
-
substrate binding, modelling and docking study,overview
-
-
-
additional information
?
-
-
the cutinase demonstrates enhanced poly(epsilon-caprolactone) hydrolysis at high temperatures and under all pH value. The cutinase shows activity on 4-nitrophenyl butyrate
-
-
-
additional information
?
-
-
the cutinase demonstrates enhanced poly(epsilon-caprolactone) hydrolysis at high temperatures and under all pH value. The cutinase shows activity on 4-nitrophenyl butyrate
-
-
-
additional information
?
-
-
the cutinase demonstrates enhanced poly(epsilon-caprolactone) hydrolysis at high temperatures and under all pH value. The cutinase shows activity on 4-nitrophenyl butyrate
-
-
-
additional information
?
-
-
the enzyme catalyzes the transesterification of triolein and methanol, overview
-
-
-
additional information
?
-
Thermobifida fusca DSM44342
-
substrate binding, modelling and docking study,overview
-
-
-
additional information
?
-
Thermobifida cellulosilytica DSM44535
-
substrate binding, modelling and docking study, overview
-
-
-
additional information
?
-
Botryotinia fuckeliana Pser.: Fr.
-
constitutive enzyme
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
constitutive enzyme
-
-
-
additional information
?
-
-
induced by cutin
-
-
-
additional information
?
-
-
Cutinase is known for its hydrolytic activity for a variety of esters ranging from soluble p-nitrophenyl esters to insoluble long-chain triglycerides. The hydrolytic activity of cutinase, especially on p-nitrophenyl esters of fatty acids, is extremely sensitive to fatty acid chain length.
-
-
-
additional information
?
-
-
cutinase is an esterase, whose active site, located at the middle of a sharp turn between beta-strand and alpha-helix, is composed by the triad Ser120, Asp175 and His188
-
-
-
additional information
?
-
-
the enzyme exhibits a broad substrate specificity against plant cutin, synthetic polyesters, insoluble triglycerides, and soluble esters
-
-
-
additional information
?
-
Botryotinia fuckeliana Pser.: Fr.
-
constitutive enzyme
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ba2+
-
activates isozyme Tfu 0883 7% at 1 mM
Co2+
-
activates 7% at 1 mM
Co2+
-
activates isozyme Tfu 0882 4% and isozyme Tfu 0883 24% at 1 mM
Mg2+
-
activates isozyme Tfu 0883 11% at 1 mM
Mn2+
-
activates 36% at 1 mM
Ni2+
-
activates isozyme Tfu 0882 5% and isozyme Tfu 0883 7% at 1 mM
Mn2+
-
activates isozyme Tfu 0882 9% and isozyme Tfu 0883 24% at 1 mM
additional information
-
EDTA has no effect on enzyme activity at 10 mM
additional information
-
Ba2+ and EDTA have no effect on enzyme activity at 1 mM and 10 mM, respectively
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(O,O)-diethyl-(3,5,6-trichloro)-2-pyridylphosphorothioate
-
1 mM, 90% inhibition
2,3,5-trichloropyridine-6-(O-methyl-O-n-butyl)-phosphate ester
-
i.e. MAT 9564
2-[(ethylsulfanyl)methyl]phenyl hydrogen methylcarbonimidate
-
-
3-(4-mercaptobutylthio)-1,1,1-trifluoro-2-propanone
-
-
3-n-octylthio-1,1,1-trifluoro-2-propanone
-
-
3-phenethylthio-1,1,1-trifluoropropan-2-one
-
-
3-phenylthio-1,1,1-trifluoropropan-2-one
-
-
4-nitrophenyl P-methyl-N-octylphosphonamidoate
-
-
alkylboronic acids
-
-
ANS
P00590
binds strongly to native cutinase as a noncompetitive inhibitor with up to 5 ANS per cutinase molecule. The first ANS molecule stabilizes cutinase. The last 4 ANS molecules decrease Tm by up to 7C
Ba2+
-
inhibits isozyme Tfu 0882 7%
Ca2+
-
inhibits 6% at 1 mM
Ca2+
-
inhibits isozyme Tfu 0882 28% and isozyme Tfu 0883 18% at 1 mM
chlorpyrifos-methyl
-
upon chloroperoxidase oxidation, chlorpyrifos-methyl shows a very strong cutinase inhibition as compared to the corresponding oxon standard
chlorpyrifos-methyl oxon
-
-
Cr3+
-
inhibits 72% at 1 mM
Cr3+
-
inhibits isozyme Tfu 0882 89% and isozyme Tfu 0883 90% at 1 mM
Cu2+
-
inhibits 17% at 1 mM
D-glucose
A8QPD8, -
Thcut1 mRNA is repressed by glucose
Diethyl p-nitrophenyl phosphate
-
covalent
Diethyl p-nitrophenyl phosphate
-
; E600
diethyl-p-nitrophenyl phosphate
-
-
diisopropyl fluorophosphate
-
102 nM, 90% inhibition
diisopropyl fluorophosphate
-
1 mM, 90% inhibition
diisopropyl fluorophosphate
Fusarium roseum
-
0.025 mM, complete inhibition
diisopropyl fluorophosphate
-
-
Fe2+
-
inhibits 54% at 1 mM
-
Fe2+
-
inhibits isozyme Tfu 0882 45% and isozyme Tfu 0883 36% at 1 mM
-
glycerol
-
inhibits the transesterification activity of the cutinase after 10 min of incubation
Hg2+
-
inhibits completely at 1 mM
Hg2+
-
inhibits isozyme Tfu 0882 and isozyme Tfu 0883 completely at 1 mM
Mg2+
-
inhibits 11% at 1 mM
Ni2+
-
inhibits 7% at 1 mM
O-(4-nitrophenyl) S-octyl methylphosphonothioate
-
-
O-methyl-O-(p-nitrophenyl)octylphosphonate
-
-
O-octyl-O-(p-nitrophenyl)ethylphosphonate
-
-
O-octyl-O-(p-nitrophenyl)methylphosphonate
-
-
oxidized malathion
-
oxidized malathion, contrarily to malaoxon, reveals cutinase inhibition
Paraoxon
Fusarium roseum
-
0.1 mM, complete inhibition
paraoxon-methyl
-
-
Pb2+
-
inhibits 39% at 1 mM
Pb2+
-
inhibits isozyme Tfu 0882 48% and isozyme Tfu 0883 52% at 1 mM
Phenylboronic acid
-
5 mM, 63% inhibition, competitive
sodium bis(2-ethylhexyl)ester sulfosuccinic acid
-
pseudo-competitive inhibitor
sodium dioctyl sulfosuccinate
-
-
Sodium dodecyl sulfate
-
competitive, detailed study of interaction with enzyme. At molar ratio of SDS:enzyme of about 10, formation of aggregates which include more than one protein molecule. At higher concentration of SDS, denaturation of protein, denatured state of enzyme is unusually compact
Zn2+
-
inhibits 18% at 1 mM
Zn2+
-
inhibits isozyme Tfu 0882 50% and isozyme Tfu 0883 56% at 1 mM
Mg2+
-
inhibits isozyme Tfu 0882 34%
additional information
-
except for methomyl no significant effects of chloroperoxidase oxidation on the inhibition strength of insecticidal carbamates can be detected. No inhibition by malathion and malaoxon
-
additional information
-
crystallization and preliminary X-ray analysis of cutinase-inhibitor complexes
-
additional information
-
inhibition by organophosphate pesticides. Carbamate pesticides reveal an efficient cutinase inhibitor effect, though less potent than the organophosphates
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
N,N-diethyl-2-phenylacetamide
-
increases hydrolysis rates of semi-crystalline poly(ethylene terephthalate) films and fabrics for cutinase. The outer layers of the polymer will be better exposed to the enzyme in the presence of N,N-diethyl-2-phenylacetamide. Overall enhancement in colour depth of 300%
olive oil
A8QPD8, -
Thcut1 mRNA is strongly induced in vitro by olive oil
-
sodium dioctyl sulfosuccinate
-
in the absence of surfactant the S54D variant catalytic activity is similar to that of the wild type cutinase, whereas L153Q and T179C variants show a lower activity
16-hydroxyhexadecanoic acid
A8QPD8, -
is strongly induced in vitro by cutin monomer 16-hydroxyhexadecanoic acid
additional information
-
pseudo-activation in presence of sodium bis(2-ethylhexyl)ester sulfosuccinic acid and hexadecyltrimethyl-ammoniumbromide
-
additional information
-
absence of interfacial activation
-
additional information
-
water:surfactant molar rate has a marked influence on the enzyme activity, with the best results in the range between 5 and 8. The use of detergents improves the reaction yield during wetting of cotton fibers. Increase in the stereoselectivity of the primary hydroxyl group acylation is obtained through the preincubation of the enzyme in the presence of the substrate diol 1, there is no correlation with the incubation time
-
additional information
-
no activation in the presence of Triton X-100 due to the absence of a lid covering the active site pocket
-
additional information
-
Pichia pastoris expressing the native non-tagged cutinase exhibits about 2- and 3fold higher values of protein amount and cutinase activity in the culture supernatant, respectively, than those containing the C-terminal tagged cutinase
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00067
-
4-nitrophenyl acetate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
0.00496
-
4-nitrophenyl acetate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
0.00021
-
4-nitrophenyl butyrate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
0.00126
-
4-nitrophenyl butyrate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
0.00136
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.00196
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.0025
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.00896
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.029
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.272
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, recombinant cutinase FspC
0.505
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, isozyme Tfu 0883
0.673
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, isozyme Tfu 0882
0.00029
-
4-nitrophenyl hexanoate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
0.0015
-
4-nitrophenyl hexanoate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
0.21
-
4-nitrophenyl hexanoate
-
-
0.86
-
4-nitrophenyl hexanoate
-
cutinase II
0.89
-
4-nitrophenyl hexanoate
-
cutinase I
0.59
-
4-nitrophenyl octanoate
-
cutinase II
0.88
-
4-nitrophenyl octanoate
-
cutinase I
1.7
-
4-nitrophenyl octanoate
-
-
4e-05
-
4-nitrophenyl valerate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
0.33
-
p-nitrophenyl acetate
A8QPD8, -
-
0.000121
-
p-nitrophenyl butyrate
P00590
20C, pH 8, no additive
0.2
-
p-nitrophenyl butyrate
-
T179C mutant, sodium dioctyl sulfosuccinate concentration = 0 mM
0.31
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0 mM
0.33
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 0 mM
0.35
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 0 mM
0.48
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 0.5 mM
0.49
-
p-nitrophenyl butyrate
-
T179Cmutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM
0.57
-
p-nitrophenyl butyrate
A8QPD8, -
-
0.66
-
p-nitrophenyl butyrate
-
T179C mutant, sodium dioctyl sulfosuccinate concentration = 1 mM
0.69
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM
0.72
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 1 mM
0.74
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM
0.85
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 1 mM
1.08
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 2 mM
1.09
-
p-nitrophenyl butyrate
-
T179C mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM
1.12
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 1.5 mM
1.14
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM
1.31
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1 mM
1.56
-
p-nitrophenyl butyrate
-
T179C mutant, sodium dioctyl sulfosuccinate concentration = 2 mM
1.74
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 2 mM
3.57
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM
4.33
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 2 mM
0.085
-
p-nitrophenyl palmitate
A8QPD8, -
-
0.82
-
p-nitrophenyl valerate
A8QPD8, -
-
0.9
-
p-Nitrophenylacetate
-
-
3
-
p-Nitrophenylacetate
-
without surfactant
6.8
-
p-Nitrophenylacetate
-
cutinase I
9.7
-
p-Nitrophenylacetate
-
cutinase II
0.27
-
p-nitrophenylbutanoate
-
-
0.35
-
p-nitrophenylbutanoate
-
cutinase I
0.35
-
p-nitrophenylbutanoate
-
-
0.47
-
p-nitrophenylbutanoate
-
without surfactant
0.75
-
p-nitrophenylbutanoate
-
cutinase II
1.23
-
p-nitrophenylbutanoate
Q8TGB8
36C, wild-type
1.45
-
p-nitrophenylbutanoate
Q8TGB8
36C, mutant H173L
1.5
-
p-nitrophenylbutanoate
Q8TGB8
36C, mutant S103T
1.68
-
p-nitrophenylbutanoate
Q8TGB8
36C, mutant S103A
0.36
-
p-nitrophenyldecanoate
-
cutinase II
0.48
-
p-nitrophenyldecanoate
-
cutinase I
3.98
-
p-nitrophenyldecanoate
-
-
0.45
-
p-nitrophenyldodecanoate
-
cutinase II
0.56
-
p-nitrophenyldodecanoate
-
cutinase I
3.55
-
p-nitrophenyldodecanoate
-
-
4.54
-
p-nitrophenylhexadecanoate
-
-
2.27
-
p-nitrophenyltetradecanoate
-
-
0.00148
-
4-nitrophenyl valerate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
additional information
-
additional information
-
Km-values in presence of surfactants
-
additional information
-
additional information
-
comparison of kinetics of cutinases from different organisms, overview
-
additional information
-
additional information
-
comparison of kinetics of cutinases from different organisms, overview
-
additional information
-
additional information
-
comparison of kinetics of cutinases from different organisms, overview
-
additional information
-
additional information
-
kinetic model of transesterification of triolein and methanol, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0072
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.0163
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.022
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.072
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
0.0845
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
483
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, isozyme Tfu 0882
742
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, isozyme Tfu 0883
7900
-
p-nitrophenyl butyrate
-
T179C mutant, sodium dioctyl sulfosuccinate concentration = 0 mM
9600
-
p-nitrophenyl butyrate
-
T179Cmutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM
11000
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0 mM; T179C mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM
11100
-
p-nitrophenyl butyrate
-
T179C mutant, sodium dioctyl sulfosuccinate concentration = 1 mM
11500
-
p-nitrophenyl butyrate
-
T179C mutant, sodium dioctyl sulfosuccinate concentration = 2 mM
13800
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 2 mM
14700
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 0 mM
15200
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 1.5 mM
15500
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 0.5 mM
15600
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 0 mM
16000
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM
17400
-
p-nitrophenyl butyrate
-
wild type, sodium dioctyl sulfosuccinate concentration = 1 mM
17900
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1 mM
24200
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 1 mM
24700
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 0.5 mM
25200
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM
25800
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 1.5 mM
28500
-
p-nitrophenyl butyrate
-
L153Q mutant, sodium dioctyl sulfosuccinate concentration = 2 mM
32100
-
p-nitrophenyl butyrate
-
S54D mutant, sodium dioctyl sulfosuccinate concentration = 2 mM
230
-
p-Nitrophenylacetate
-
without surfactant
0.25
-
p-nitrophenylbutanoate
Q8TGB8
36C, mutant H173L; 36C, mutant S103T
0.66
-
p-nitrophenylbutanoate
Q8TGB8
36C, wild-type
1.5
-
p-nitrophenylbutanoate
Q8TGB8
36C, mutant S103A
539
-
p-nitrophenylbutanoate
-
without surfactant
837
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, recombinant cutinase FspC
additional information
-
additional information
-
turnover numbers in presence of surfactants
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.67e-06
-
4-nitrophenyl acetate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
4735
4.217e-05
-
4-nitrophenyl acetate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
4735
4.3e-07
-
4-nitrophenyl butyrate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
4739
5.82e-05
-
4-nitrophenyl butyrate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
4739
0.0053
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
4739
0.0093
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
4739
0.0162
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
4739
0.029
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
4739
0.0567
-
4-nitrophenyl butyrate
-
pH 7.5, 25C
4739
150
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, isozyme Tfu 0883
4739
700
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, isozyme Tfu 0882
4739
3200
-
4-nitrophenyl butyrate
-
pH 8.0, 60C, recombinant cutinase FspC
4739
2.3e-06
-
4-nitrophenyl hexanoate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
19383
2.3e-05
-
4-nitrophenyl hexanoate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
19383
1.017e-05
-
4-nitrophenyl valerate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
204888
5.3e-05
-
4-nitrophenyl valerate
-
in 14.5 mM Tris-HCl buffer, pH 7.5, 0.75% glycerol
204888
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.44
-
sodium dioctyl sulfosuccinate
-
L153Q mutant
0.46
-
sodium dioctyl sulfosuccinate
-
T179C mutant
0.72
-
sodium dioctyl sulfosuccinate
-
wild type
1.38
-
sodium dioctyl sulfosuccinate
-
S54D mutant
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.004
-
A8QPD8, -
4-nitrophenyl palmitate, esterase activity of the recombinant THCUT1 protein against different chromogenic substrates
0.12
-
A8QPD8, -
in 1% olive oil, esterase activity values decreased after 4 h of incubation and are 1.8, 1.1, and 0.12 micromol/min/mg against 4-nitrophenyl palmitate, at 4, 8 and 24 h
0.25
-
A8QPD8, -
4 h cultures against 4-nitrophenyl palmitate
0.4
-
A8QPD8, -
4-nitrophenyl valerate, esterase activity of the recombinant THCUT1 protein against different chromogenic substrates
0.74
-
A8QPD8, -
4-nitrophenyl butyrate, esterase activity of the recombinant THCUT1 protein against different chromogenic substrates
0.85
-
A8QPD8, -
in 1% olive oil, esterase activity values decreased after 4 h of incubation and are 11.4, 5.4, and 0.85 micromol/min/mg against 4-nitrophenyl butyrate, at 4, 8 and 24 h
1.1
-
A8QPD8, -
in 1% olive oil, esterase activity values decreased after 4 h of incubation and are 1.8, 1.1, and 0.12 micromol/min/mg against 4-nitrophenyl palmitate, at 4, 8 and 24 h
1.2
-
A8QPD8, -
supernatant from 8 h culture on 0.2% cutin monomer 16-hydroxy-hexadecanoic acid shows slightly higher values against 4-nitrophenyl palmitate
1.3
-
A8QPD8, -
supernatant from 8 h culture on 0.05% cutin monomer 16-hydroxy-hexadecanoic acid shows slightly higher values against 4-nitrophenyl palmitate
1.8
-
A8QPD8, -
in 1% olive oil, esterase activity values decreased after 4 h of incubation and are 1.8, 1.1, and 0.12 micromol/min/mg against 4-nitrophenyl palmitate, at 4, 8 and 24 h
2.77
-
-, P41744
carrier bound CUTAB1 with tributyrin as substrate, in 40 ml of 50 mM potassium phosphate buffer, pH 5.5 containing 25% ethanol, at 8C, for 40 H
2.84
-
A8QPD8, -
4-nitrophenyl acetate, esterase activity of the recombinant THCUT1 protein against different chromogenic substrates
4.2
-
A8QPD8, -
4 h cultures against 4-nitrophenyl butyrate (6.6 and 4.2 micromol/min/mg)
5.4
-
A8QPD8, -
in 1% olive oil, esterase activity values decreased after 4 h of incubation and are 11.4, 5.4, and 0.85 micromol/min/mg against 4-nitrophenyl butyrate, at 4, 8 and 24 h
6.6
-
A8QPD8, -
4 h cultures against 4-nitrophenyl butyrate (6.6 and 4.2 micromol/min/mg)
7.5
-
A8QPD8, -
supernatant from 8 h culture on 0.2% cutin monomer 16-hydroxy-hexadecanoic acid shows higher activity values against 4-nitrophenyl butyrate
9.8
-
A8QPD8, -
supernatant from 8 h culture on 0.05% cutin monomer 16-hydroxy-hexadecanoic acid shows higher activity values against 4-nitrophenyl butyrate
10
-
-, P41744
crude extract, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
11.4
-
A8QPD8, -
in 1% olive oil, esterase activity values decreased after 4 h of incubation and are 11.4, 5.4, and 0.85 micromol/min/mg against 4-nitrophenyl butyrate, at 4, 8 and 24 h
17
-
-, P41744
1.6fold purified enzyme, with 4-nitrophenyl palmitate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic; wild-type, with 4-nitrophenyl palmitate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
30
-
-, P41744
wild-type, with tripalmitin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
40
-
-
cutinase-tryptophan,proline4, after 72 h of Saccharomyces cerevisae cultivation
41
-
-, P41744
wild-type, with methyl acetate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
45
-
Q9SZW7
at 30C
47
-
-, P41744
wild-type, with tristearin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
66
-
-, P41744
mutant A84F, with tripalmitin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
77
-
-, P41744
mutant A84F, with tristearin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
125
-
-, P41744
wild-type, with ethyl caprylate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic; wild-type, with methyl caproate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
129
-
-, P41744
wild-type, with methyl caprylate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
145
-
-, P41744
wild-type, with methyl myristate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
157
-
-, P41744
wild-type, with methyl propionate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
161
-
-, P41744
wild-type, with methyl decanoate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
170
-
-
lyophilized cutinase, pH 8.0, 30C
188
-
-, P41744
mutant A84F, with 4-nitrophenyl palmitate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
197
-
-, P41744
wild-type, with methyl laurate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
200
-
-
cutinase-tryptophan,proline2, after 72 h of Saccharomyces cerevisae cultivation
257
-
-, P41744
wild-type, with methyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
281
-
-, P41744
wild-type, with ethyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
350
-
-
wild type, after 72 h of Saccharomyces cerevisae cultivation
454
-
-, P41744
wild-type, with triolein as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
586
-
-, P41744
mutant A84F, with triolein as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
948
-
-, P41744
mutant A84F, with 4-nitrophenyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
1057
-
-, P41744
wild-type, with 4-nitrophenyl butyrate as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
1286
-
-, P41744
wild-type, with tricaprylin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
2019
-
-, P41744
mutant A84F, with tricaprylin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
3302
-
-, P41744
wild-type, with tributyrin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
4394
-
-, P41744
mutant A84F, with tributyrin as substrate, in 0.1 M Tris-HCl buffer, pH 8.0 with 0.5% (v/v) Triton X-100 and 0.1% (w/v) gum arabic
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
additional information
-
-
Assay of enzymatic activity of the fusion mutant of cutinase (RpoS-CUT) shows the same selective bioactivity as native cutinase to degrade p-nitrophenyl butyrate (PNB) but not to degrade p-nitrophenyl butyrate.
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
-
B9U443, -
and a second optimum at 8.0, but retains activity over a wide pH range
7.5
8
-
assay at
8
-
-
hydrolysis of cutin or p-nitrophenylbutyrate
8
-
B9U443, -
and a second optimum at 7.0, but retains activity over a wide pH range
8
-
-
assay at
8
-
-
assay at
8.5
10.5
-
-
8.5
-
-
study of thermal unfolding of enzyme as a function of pH-value in different buffers. At pH-optimum of 8.5, enzyme also has high thermal stability
10
-
Fusarium roseum, Fusarium solani
-
-
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
9
-
has activity superior to lipases
6
9
-
pH dependence of enzyme activity is determined between pH 6.0 and 9.0
7
9
-, P41744
at both, room temperature and 40C
8
10
-
non-tagged and the C-terminal tagged cutinases
8
11
-
pH 8.0: about 60% of maximal activity, pH 11.0: about 50% of maximal activity
8
11
Fusarium roseum
-
pH 8.0: about 40% of maximal activity, pH 11.0: about 75% of maximal activity
additional information
-
-
evaluation of temperature and pH effect on protein conformation and dynamics by study of fluorescence of the single tryptophan
additional information
-
B9U443, -
retains activity over a wide pH range
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-
displays an optimal temperature at 30C for triolein and 40C for 4-nitrophenyl butyrate
37
-
-
assay at
37
-
-
assay at
40
-
B9U443, -
activity assay at
40
-
-
displays an optimal temperature at 30C for triolein and 40C for 4-nitrophenyl butyrate
70
-
-
epsilon-caprolactone ring-opening polymerizations
70
-
-
immobilized enzyme
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
70
-
temperature dependence of enzyme activity is determined between 20 and 70C
35
45
-
non-tagged and the C-terminal tagged cutinases
60
80
-
60C: about 25% of maximal activity, 80C: less than 10% of maximal activity
additional information
-
-
evaluation of temperature and pH effect on protein conformation and dynamics by study of fluorescence of the single tryptophan
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
-
P00590
-
8.1
-
A6N6J6
calculated from sequence
8.2
-
Q8TGB8
mature form, calculated
8.4
-
Q8TGB8
precursor protein, calculated
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Botryotinia fuckeliana Pser.: Fr.
-
-
-
Manually annotated by BRENDA team
Fusarium roseum
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
Q9SZW7
in mature pollen grains within non-dehiscent anthers
Manually annotated by BRENDA team
Q9SZW7
pollen tubes that are germinating on the stigma
Manually annotated by BRENDA team
Q9SZW7
at the zone of lateral root emergence
Manually annotated by BRENDA team
Q9SZW7
CDEF1 is secreted to the extracellular space in leaves
Manually annotated by BRENDA team
additional information
-
fermentation conditions for cutinase production with a mutant of Thermobifida fusca ATCC 27730 are studied
Manually annotated by BRENDA team
additional information
A6N6J6
microconidium
Manually annotated by BRENDA team
additional information
-
microconidium
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Botryotinia fuckeliana Pser.: Fr.
-
110000 Da enzyme
-
Manually annotated by BRENDA team
Botryotinia fuckeliana Pser.: Fr.
-
40800 Da enzyme
-
Manually annotated by BRENDA team
Fusarium roseum
-
-
-
Manually annotated by BRENDA team
-
110000 Da enzyme, bound to membrane or cell wall
Manually annotated by BRENDA team
Botryotinia fuckeliana Pser.: Fr.
-
110000 Da enzyme, bound to membrane or cell wall
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Aspergillus oryzae (strain ATCC 42149 / RIB 40)
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
Fusarium solani subsp. pisi
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6982
-
B9U443, -
N-terminal fragment, electrospray ionization Q-TOF mass spectrometric analysis
12140
-
B9U443, -
C-terminal fragment, Ala(95) - His(204), electrospray ionization Q-TOF mass spectrometric analysis
12270
-
B9U443, -
C-terminal fragment, Ala(95) - His(205), electrospray ionization Q-TOF mass spectrometric analysis
18800
-
B9U443, -
around, electrospray ionization Q-TOF mass spectrometric analysis gives two masses 18726.0 Da and 18863.0 Da
20000
-
B9U443, -
SDS-PAGE
20400
-
-
cutinase II, equilibrium sedimentation
21010
-
-
after preincubation with 20 mM diethyl p-nitrophenyl phosphate, the peak at 21010 Da represents the nonmodified H204N mutant, mass spectrometry; H204N mutant, mass spectrometry; H204N site-directed mutant, mass spectrometry
21030
-
-
wild type, mass spectrometry; wild-type, mass spectrometry
21170
-
-
after preincubation with 20 mM diethyl p-nitrophenyl phosphate, the peak at 21168 Da represents the covalently modified wild-type cutinase, mass spectrometry
21700
-
-
gel filtration
21700
-
A6N6J6
calculated from sequence
21800
-
-
cutinase I and II, gel filtration
22000
-
-
molar mass found for the wild type cutinase and its mutants is identical
23400
-
-
cutinase I, equilibrium sedimentation
29000
-
A8QPD8, -
SDS-PAGE
30000
-
-
gel filtration
30100
-
-
sequence analysis
32000
-
-
recombinant isozymes Tfu 0882 and Tfu 0883, gel filtration
36400
-
-
sequence analysis
38700
-
-
sequence analysis
39200
-
-
sequence analysis
39200
-
-
sequence analysis
41200
-
-
sequence analysis
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 21800, and a smaller band of 10600 Da, cutinase II, SDS-PAGE; x * 21800, cutinase I, SDS-PAGE
?
-
x * 22000, SDS-PAGE
?
-
x * 110000, enzyme from cell wall or membrane, SDS-PAGE; x * 40800, soluble enzyme, SDS-PAGE
?
-
x * 18600, protein bands of 18600 Da and 20800 Da show cutinase activity; x * 20800, protein bands of 18600 Da and 20800 Da show cutinase activity
?
Fusarium roseum
-
x * 24300, SDS-PAGE
?
-
x * 22800, x * 24900, two differently glycosylated forms, SDS-PAGE
?
Q8TGB8
x * 20227, precursor protein, x * 18134, mature form, calculated
?
-
x * 38700, calculated from sequence
?
-
x * 30100, calculated from sequence
?
-
x * 36400, calculated from sequence
?
-
x * 41200, calculated from sequence
?
-
x * 39200, calculated from sequence
?
-
x * 39200, calculated from sequence
?
Botryotinia fuckeliana Pser.: Fr.
-
x * 110000, enzyme from cell wall or membrane, SDS-PAGE; x * 40800, soluble enzyme, SDS-PAGE
-
monomer
-
1 * 21400, cutinase I and II, SDS-PAGE
monomer
-
1 * 23000, SDS-PAGE
monomer
-
1 * 30000, SDS-PAGE
monomer
-, P41744
1 * 24000, SDS-PAGE, glycosylated CUTAB1. 1 * 20000, SDS-PAGE, non-glycosylated CUTAB1
monomer
-
crystallography
monomer
-
1 * 29000, isozymes Tfu 0882 and Tfu 0883, SDS-PAGE, 1 * 29220, isozyme Tfu 0882, sequence calculation, 1 * 28997, isozyme Tfu 0883, sequence calculation
additional information
-
cutinase isozyme structure comparisons of cutinases, overview
additional information
Thermobifida cellulosilytica DSM44535
-
cutinase isozyme structure comparisons of cutinases, overview
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
two differently glycosylated forms of 22800 and 24900 Da
glycoprotein
-
contains 5.4% carbohydrate
glycoprotein
-, P41744
the glycosylated form of CUTAB1 has a MW of 24000 Da. The non-glycosylated form of CUTAB1 has a MW of 20000 Da
additional information
Q8TGB8
sequence contains a 20 amino-acid secretory signal, one potential N-linked glycosilation site, a protein kinase C phosphorylation site, a lipase active serine and a cutinase active serine
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
to 1.75 A resolution. Alpha/beta fold hallmarked by a central beta-sheet of 5 parallel strands surrounded by 10 alpha-helices. An additional disulfide bond and a topologically favored catalytic triad (Ser126, Asp181, and His194), with a continuous and deep groove
-
crystallization of native enzyme, 27 mutant enzymes and 4 covalently inhibited complexes, crystallizes in 8 different crystal forms
-
; in the absence and in the presence of the inhibitors diethyl p-nitrophenyl phosphate (belongs to space group P21) and 3-phenethylthio-1,1,1-trifluoropropan-2-one (belongs to space group P212121), to resolutions of 2.6 and 2.3 A, respectively. Apo-cutinase, 1.9 A resolution, belongs to space group P41212 with one subunit in the asymmetric unit with unit cell parameters a = 60, b = 60, c = 86 A, respectively. The catalytic triad (Ser136, Asp191, and His204) adopts an unusual configuration with the putative essential histidine His204 swung out of the active site into a position where it is unable to participate in catalysis, with the imidazole ring 11 A away from its expected position
-
crystallization and preliminary X-ray analysis of cutinase-inhibitor complexes, resolution beyond 1.6 A
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
B9U443, -
retains activity over a wide pH range
additional information
-
-
the enzyme is unstable and functions poorly at high temperatures as well as at acidic pH conditions, biophysical parameters of cutinase as a function of pH, overview
additional information
-
-
the enzyme is unstable and functions poorly at high temperatures as well as at acidic pH conditions, biophysical parameters of cutinase as a function of pH, overview
additional information
-
-
the enzyme is unstable and functions poorly at high temperatures as well as at acidic pH conditions, biophysical parameters of cutinase as a function of pH, overview
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
-
-
pH 8, in a water bath with agitation, half-life without additives: 45 days, half-life with 7.5% N,N-dimethylacetamide: 48 days, half-life with 15% N,N-dimethylacetamide: 159 days, half-life with 25% N,N-dimethylacetamide: 4 days, half-life with 50% N,N-dimethylacetamide: 0.4 day, half-life in presence of 25% glycerol: 134 days, half-life with 25% sorbitol: 113 days, half-life with 25% xylitol: 31 days, half-life in presence of 50% ethylene glycol: 29 days, half-life in presence of 50% polyethylene glycol: 23 days
38.5
-
-
50% unfolding at pH 10.9; 50% unfolding at pH 10.9, presence of 0.5 M trehalose
40
60
-
thermostability of Fusarium solani pisi cutinase is evaluated at both 40 and 60C. It exhibits a similar initial increase at 40C, but a simple exponential decay at 60C. 50% of activity retained after 85 h at 40C or after 5 min at 60C
40
-
P00590
0.08 mM cutinase, pH 8.0, no additive, Tm-value is 40.3C
40
-
-, P41744
retains 84% of its initial activity at 40C and pH 8.0 for 5 days
42
-
-
50% unfolding at pH 10.5; 50% unfolding at pH 10.5, presence of 0.5 M trehalose
48
-
P00590
0.08 mM cutinase or 0.005 mM cutinase, pH 8.0, 2 M urea, Tm-value 47.5C
49
-
-
at pH 4.5, cutinase unfolds with a Tm of 49.3C. point. The Tm value increases 7.2 C in the presence of 1 M of trehalose
49.8
-
-
50% unfolding at pH 4.5; 50% unfolding at pH 4.5, presence of 0.5 M trehalose
50
-
B9U443, -
enzyme retains 80% of its activity after 20 h incubation at 50C, but residual activity decreases sharply at 60C
52
-
P00590
0.06 mM cutinase, pH 8.0, Tm-value is 52.0C
52.6
-
-
50% unfolding at pH 9.2; 50% unfolding at pH 9.2, presence of 0.5 M trehalose
55
-
P00590
0.08 mM cutinase, pH 8.0, 20 mM 0.5 M L-Arg, Tm-value
56
-
P00590
0.005 mM cutinase, pH 8.0, 20 mM 0.5 M L-Arg, Tm-value is 56.4C
56
-
-
melting temperature. Dramatic loss in activity at 40 C, 40% drop at 30 C, continuous decline in activity as a function of increasing temperature
57
-
P00590
0.04 mM cutinase, pH 8.0, Tm-value is 57.3C
57
-
-
Tm-value in prresence of 1 mM trehalose
59
-
P00590
0.0001 mM cutinase, pH 8.0, Tm-value is 59.3C; 0.005 mM cutinase, pH 8.0, Tm-value is 59.3C
59
-
-
melting temperature. Maintains a high level of activity at 40 C
additional information
-
-
study of thermal unfolding of enzyme as a function of pH-value in different buffers. At pH-optimum of 8.5, enzyme also has high thermal stability
additional information
-
P00590
ANS binds strongly to native cutinase as a noncompetitive inhibitor with up to 5 ANS per cutinase molecule. The first ANS molecule stabilizes cutinase. The last 4 ANS molecules decrease Tm-value by up to 7C
additional information
-
-
trehalose delays thermal unfolding, thus increasing the temperature at the mid-point of unfolding by 7.2C
additional information
-
-
thermal stability of Humicola insolens cutinase in aqueous SDS. SDS stabilizes noticeably against irreversible aggregation
additional information
-
-
presence of hexanol and the low water content lead to the enzyme stabilization in the interior of the micelles, increasing its thermostability
additional information
-
Q9SZW7
activity is reduced to less than 10% of the maximum activity by heating for 10 min at 99C
additional information
-
-
the enzyme is unstable and functions poorly at high temperatures as well as at acidic pH conditions, differential scanning calorimetry thermograms of cutinase, overview
additional information
-
-
the enzyme is unstable and functions poorly at high temperatures as well as at acidic pH conditions, differential scanning calorimetry thermograms of cutinase, overview
additional information
-
-
the enzyme is unstable and functions poorly at high temperatures as well as at acidic pH conditions, differential scanning calorimetry thermograms of cutinase, overview
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ANS binds strongly to native cutinase as a noncompetitive inhibitor with up to 5 ANS per cutinase molecule. The first ANS molecule stabilizes cutinase. The last 4 ANS molecules decrease Tm by up to 7C
P00590
encapsulation of enzyme in bis(2-ethylhexyl) sodium sulfosuccinate reverse micelles induces unfolding at room temperature, presence of 1-hexanol delays or even prevents unfolding
-
half-life time of cutinase at 35C, pH 8, in a water bath with agitation, increases by 3.5fold with the addition of 15% N,N-dimethylacetamide and by 3fold with 1 M glycerol
-
in bis(2-ethylhexyl) sodium sulfosuccinate reverse micelles, enzyme unfolds with mono-exponential rates, indicating a two-state process. After unfolding in reverse micelles, enzyme is less destabilised than in guanidinium hydrochloride-denatured state, which is supported by fluorescence data. NMR studies indicate a molten globule structure
-
micelle-forming short-chain phospholipids significantly reduce cutinase stability (both below and above the critical micelle concentration) and rates of folding (only above critical micelle concentration), trapping cutinase in an inactive state which only regains activity over hours to days, rather than the few seconds required for refolding in the absence of detergent. Destabilization decreases with increasing chain length, and increases with critical micelle concentration, indicating that monomers and micelles cooperate in destabilizing cutinase
-
stable in presence of hexadecyltrimethyl-ammoniumbromide
-
trehalose delays thermal unfolding, thus increasing the temperature at the mid-point of unfolding by 7.2C
-
unfolding of enzyme induced by guanidinium hydrochloride shows a stable intermediate, molten globule
-
unstable in presence of sodium bis(2-ethylhexyl)ester sulfosuccinic acid, unfolding of protein structure
-
in bis(2-ethylhexyl) sodium sulfosuccinate reverse micelles, enzyme unfolds with mono-exponential rates, indicating a two-state process. After unfolding in reverse micelles, enzyme is less destabilised than in guanidinium hydrochloride-denatured state, which is supported by fluorescence data. NMR studies indicate a molten globule structure
-
Using RNA polymerase sigma factor (RpoS) or glutathione transferase as fusion expression partners, the solubility of cutinase significantly increases.
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
vinyl acetate
-
has a stabilizing effect on the enzyme activity
additional information
-
hexanol has a stabilizing effect on the enzyme activity in reverse micelles
additional information
-
stability of the cutinase in different organic solvents. The cutinase is incubated with 75% v/v of organic solvent in assay buffer at 20C for 18 h, overview
additional information
-
stability of the cutinase in different organic solvents. The cutinase isozymes are incubated with 75% v/v of organic solvent in assay buffer at 20C for 18 h, overview
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
22C, high stability
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
wild-type and mutants purified by ultrafiltration, wild-type further purified by hydrophobic interaction chromatography and ammonium sulfate precipitation, 1.6fold with a yield of 70%
-, P41744
on Ni2+ affinity column
Q9SZW7
by immobilized metal affinity chromatography exploiting a C-terminal His-tag
B9U443, -
optimization and evaluation of foam fractionation as purification method to purify an extracellular cutinase from untreated supernatant of mycelium of submerged cultures of the basidiomycete Coprinopsis cinerea as a model enzyme, detailed overview
-
; by nickel-affinity chromatography
-
by centrifugation, ultrafiltration and on Ni2+ column
-
non-tagged and the C-terminal tagged cutinases, by diafiltration
-
osmotic shock, acid precipitation, dialysis and two sequential anion exchange chromatographic steps, followed by a final dialysis and subsequently freeze-dried. Cutinase purity is confirmed by 12.5% SDS-PAGE
-
; by ion-exchange chromatography
-
affinity purification
-
His-tagged protein, expressed in Pichia pastoris
Q8TGB8
recombinant cutinase FspC from Bacillus subtilis
-
study of enzyme partition in a 20% polyethylene glykol/15% phosphate two-phase system. Specific interaction of butyrate to the active site of enzyme, enzyme-butyrate complex is over two times the size of the free enzyme
-
purification of his-tagged protein using Ni2+-affinity chromatography
-
purification includes cationic Expanded Bed Adsorption (EBA) followed by hydrophobic interaction chromatography (HIC)
-
recombinant cutinase isozymes Tfu 0882 and Tfu 0883 from Escherichia coli strain BL21(DE3)
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
PCR product digested with EcoRI and NotI, and ligated in corresponding sites of the linearized pPICZalphaA vector. Transformation of Escherichia coli DH5alpha cells. Wild-type and mutants heterologously expressed in Pichia pastoris X-33 cells under the control of the methanol-inducible AOX1 promoter
-, P41744
transgenic Arabidopsis plants expressing GFP-CDEF1 fusion protein driven by the 35S promoter. CDEF1 expressed with a 6 x His tag at its C-terminus in suspension cultures of BY-2 tobacco cells
Q9SZW7
cloned and expressed in Trichoderma reesei
B9U443, -
amplified DNA fragments cloned into the pGEM-T vector, cut1 isolated from a lambdaEMBL3 genomic library
A6N6J6
cloned in pMa5-L and overexpressed in Escherichia coli
-
cutinase S120A is expressed in Escherichia coli BL21(DE3)
P00590
expressed in Escherichia coli Origami B(DE3); expressed in Escherichia coli strain Origami B(DE3)
-
expressed in Pichia pastoris
-
expression in Escherichia coli WK-6 strain
-
into pPICZalphaA with the Saccharomyces cerevisiae alpha-factor signal sequence and methanol-inducible alcohol oxidase promoter, additional sequences of the c-myc epitope and (His)6-tag of the vector fused to the C-terminus of cutinase, while the other expression vector is constructed without any additional sequence. Expressed in Pichia pastoris strain X-33
-
over-expression of recombinant cutinase cloned in pMac5-8 and its variants (S54D, L153Q and T179C) are performed with Escherichia coli WK6 strain
-
overexpression of recombinant cutinase and its variants is performed with strain Escherichia coli WK-6 (delta(lac-pro) galE strA F'[lacIq ZdeltaM15 proAB+])
-
cloned and overexpressed in Escherichia coli
-
expressed in Escherichia coli strain Origami B(DE3); mutant H204N overexpressed in Escherichia coli Origami B(DE3)
-
expressed in Pichia pastoris
A8QPD8, -
expression of cutinase FspC in Bacillus subtilis
-
overexpression in Escherichia coli strain WK-6
-
recombinant expression of cutinase in Saccharomyces cerevisiae strain SU50
-
Expression of aggregation-prone cutinase protein using RNA polymerase sigma factor (RpoS) or glutathione transferase as fusion partners in Escherichia coli strain BL21 (DE3).
-
expression of cutinase isozymes Tfu 0882 and Tfu 0883 in Escherichia coli strain BL21(DE3)
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
CDEF1 gene is not expressed in leaves of wild-type plants. Is expressed at low levels in roots
Q9SZW7
CDEF1 gene is expressed in leaves of transgenic 35S::CDEF1 plants. CDEF1 gene is highly expressed in mature pollen
Q9SZW7
mutants carrying a DELTActf1 loss-of-function allele grown on inducing substrates fail to activate extracellular cutinolytic activity and expression of the cut1 gene, encoding a putative cutinase
A6N6J6
the zinc finger transcription factor Ctf1 from Fusarium oxysporum mediates efficient transcriptional activation of cut1 and lip1 on fatty acid substrates, but not on glucose. The Ctf1 transcription factor, a functional orthologue of CTF1alpha, controls expression of cutinase genes and virulence. Strains harbouring a ctf1C allele in which the ctf1 coding region is fused to the strong constitutive Aspergillus nidulans gpdA promoter show increased induction of cutinase activity and gene expression. Higher levels of cut1 transcript in mycelia of the wild-type grown on apple cutin than in those grown on glucose. Activation of cut1 by apple cutin is even more pronounced in the ctf1C strain than in the wild-type. Low expression of cut1 in roots 10 days after inoculation of tomato plants
A6N6J6
mutants carrying a DELTActf1 loss-of-function allele grown on inducing substrates fail to activate extracellular cutinolytic activity and expression of the cut1 gene, encoding a putative cutinase
-
-
the zinc finger transcription factor Ctf1 from Fusarium oxysporum mediates efficient transcriptional activation of cut1 and lip1 on fatty acid substrates, but not on glucose. The Ctf1 transcription factor, a functional orthologue of CTF1alpha, controls expression of cutinase genes and virulence. Strains harbouring a ctf1C allele in which the ctf1 coding region is fused to the strong constitutive Aspergillus nidulans gpdA promoter show increased induction of cutinase activity and gene expression. Higher levels of cut1 transcript in mycelia of the wild-type grown on apple cutin than in those grown on glucose. Activation of cut1 by apple cutin is even more pronounced in the ctf1C strain than in the wild-type. Low expression of cut1 in roots 10 days after inoculation of tomato plants
-
-
cutinase genes show four differential expression patterns, indicating regulatory sub- and neo-functionalization
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A84F
-, P41744
mutation in the small helical flap, significantly increases the activity towards longer chain substrates like 4-nitrophenyl palmitate
I183A
-, P41744
mutation in the hydrophobic binding loop, drastically reduces the overall activity
L181A
-, P41744
mutation in the hydrophobic binding loop, drastically reduces the overall activity
A164E
-
amino acid substitution, 74% of wild-type activity
A164R
P00590
41% of the activity of the wild-type enzyme
A185L
P00590
96% of the activity of the wild-type enzyme
A185T
-
amino acid substitution, 142% of wild-type activity
A195S
P00590
38% of the activity of the wild-type enzyme
A199C
-
comparative structural analysis of native enzyme and mutant enzymes
A199C
P00590
no activity
A29S
-
64% of the activity of the wild-type enzyme
A79G
P00590
50% of the activity of the wild-type enzyme
A85F
-
crystallizes in a form different from the native enzyme
A85F
-
optimal activity with triglyceride anolgues shifts towards slightly longer acyl ester chains
A85F
P00590
136% of the activity of the wild-type enzyme
A85F/G82A
-
optimal activity with triglyceride anolgues shifts towards slightly longer acyl ester chains
A85W
-
optimal activity with triglyceride anolgues shifts towards slightly longer acyl ester chains
A85W
P00590
5% of the activity of the wild-type enzyme
D111N
P00590
39% of the activity of the wild-type enzyme
D134S
P00590
37% of the activity of the wild-type enzyme
D83S
P00590
62% of the activity of the wild-type enzyme
E201K
P00590
54% of the activity of the wild-type enzyme
G192Q
P00590
44% of the activity of the wild-type enzyme
G26A
P00590
32% of the activity of the wild-type enzyme
G26P
-
amino acid substitution, 68% of wild-type activity
G41A
-
amino acid substitution, 76% of wild-type activity
G82A
-
mutation has no influence on enzymatic properties
I183F
P00590
25% of the activity of the wild-type enzyme
I204K
P00590
66% of the activity of the wild-type enzyme
I24S
P00590
4% of the activity of the wild-type enzyme
K140D
-
amino acid substitution, 39% of wild-type activity
K151M
-
amino acid substitution, 22% of wild-type activity
K151R
P00590
29% of the activity of the wild-type enzyme
K168L
P00590
83% of the activity of the wild-type enzyme
K65P
-
amino acid substitution, 99% of wild-type activity
L114C
-
comparative structural analysis of native enzyme and mutant enzymes
L114Y
P00590
20% of the activity of the wild-type enzyme
L153A
-
amino acid substitution, 111% of wild-type activity
L153Q
-
L153Q mutation also reduces the development of hydrophobic solvent accessible patches
L153Q
-
amino acid substitution, 145% of wild-type activity
L182A
-
shows the one- and two-fold higher ability to biodegrade aliphatic polyamide substrates. Activity with polyethylene terephthalate fibers is 5.3fold higher than wild-type enzyme, activity with polyamide 6,6 fiber is 119% of wild-type activity
L182W
P00590
19% of the activity of the wild-type enzyme
L189A
-
activity with polyethylene terephthalate fibers is 78% of wild-type enzyme, activity with polyamide 6,6 fiber is 94% of wild-type activity
L189F
-
comparative structural analysis of native enzyme and mutant enzymes
L189F
P00590
109% of the activity of the wild-type enzyme
L81A
-
activity with polyethylene terephthalate fibers is 4fold higher than wild-type enzyme, activity with polyamide 6,6 fiber is 98% of wild-type activity
L81G/L182G
-
comparative structural analysis of native enzyme and mutant enzymes
L99K
P00590
78% of the activity of the wild-type enzyme
N161D
P00590
63% of the activity of the wild-type enzyme
N172K
-
comparative structural analysis of native enzyme and mutant enzymes
N172K
P00590
45% of the activity of the wild-type enzyme
N172K/R196E
-
comparative structural analysis of native enzyme and mutant enzymes; crystallizes in a form different from the native enzyme
N33S
P00590
74% of the activity of the wild-type enzyme
N84A
-
comparative structural analysis of native enzyme and mutant enzymes
N84A
-
26.5% of the activity of the wild-type enzyme with p-nitrophenylbutanoate as substrate
N84A
P00590
5% of the activity of the wild-type enzyme
N84A
-
activity with polyethylene terephthalate fibers is 1.7fold higher than wild-type enzyme, activity with polyamide 6,6 fiber is 93% of wild-type activity
N84D
-
crystallizes in a form different from the native enzyme
N84D
-
0.16% of the activity of the wild-type enzyme with p-nitrophenylbutanoate as substrate
N84D
P00590
no activity
N84L
-
comparative structural analysis of native enzyme and mutant enzymes
N84L
-
3.0% of the activity of the wild-type enzyme with p-nitrophenylbutanoate as substrate
N84L
P00590
5% of the activity of the wild-type enzyme
N84W
-
comparative structural analysis of native enzyme and mutant enzymes
N84W
-
0.11% of the activity of the wild-type enzyme with p-nitrophenylbutanoate as substrate
Q121L
-
comparative structural analysis of native enzyme and mutant enzymes
R156E
P00590
79% of the activity of the wild-type enzyme
R156K
P00590
115% of the activity of the wild-type enzyme
R156K
-
amino acid substitution, 151% of wild-type activity
R156L
-
comparative structural analysis of native enzyme and mutant enzymes
R156L
P00590
71% of the activity of the wild-type enzyme
R156N
-
amino acid substitution, 89% of wild-type activity
R158L
-
amino acid substitution, 75% of wild-type activity
R17E
P00590
34% of the activity of the wild-type enzyme
R17E/N172K
-
comparative structural analysis of native enzyme and mutant enzymes
R17N
P00590
31% of the activity of the wild-type enzyme
R17S
-
amino acid substitution, 69% of wild-type activity
R196A
-
amino acid substitution, 75% of wild-type activity
R196E
-
crystallizes in a form different from the native enzyme
R196E
P00590
45% of the activity of the wild-type enzyme
R196E
-
amino acid substitution, 21% of wild-type activity
R196K
P00590
38% of the activity of the wild-type enzyme
R196L
P00590
44% of the activity of the wild-type enzyme
R208A
P00590
64% of the activity of the wild-type enzyme
R78L
P00590
49% of the activity of the wild-type enzyme
R78N
P00590
34% of the activity of the wild-type enzyme
R88A
P00590
39% of the activity of the wild-type enzyme
R96N
P00590
57% of the activity of the wild-type enzyme
S120A
P00590
no activity
S120A
P00590
the mutant enzyme casries a 15 amino acid pro-peptide. The pro-peptide is affected by the presence of the micellar substrate
S120C
-
comparative structural analysis of native enzyme and mutant enzymes
S129C
-
comparative structural analysis of native enzyme and mutant enzymes
S213C
-
comparative structural analysis of native enzyme and mutant enzymes
S42A
-
comparative structural analysis of native enzyme and mutant enzymes
S42A
-
0.22% of the activity of the wild-type enzyme with p-nitrophenylbutanoate as substrate
S42A
P00590
no activity
S54D
-
S54D mutant of cutinase is significantly more resistant to sodium dioctyl sulfosuccinate denaturation than the wild type
S54D
-
amino acid substitution, 79% of wild-type activity
S54E
P00590
34% of the activity of the wild-type enzyme
S54E
-
amino acid substitution, 83% of wild-type activity
S54K
P00590
96% of the activity of the wild-type enzyme
S54W
-
mutation has no influence on enzymatic
S54W
P00590
89% of the activity of the wild-type enzyme
S57D
-
amino acid substitution, 61% of wild-type activity
S61D
-
amino acid substitution, 83% of wild-type activity
S82R
P00590
50% of the activity of the wild-type enzyme
S92C
-
comparative structural analysis of native enzyme and mutant enzymes
T144C
-
comparative structural analysis of native enzyme and mutant enzymes
T144C
P00590
54% of the activity of the wild-type enzyme
T167L
P00590
54% of the activity of the wild-type enzyme
T173K
P00590
119% of the activity of the wild-type enzyme
T179C
-
T179C mutation located close to the active centre and to disulfide bond Cys171-Cys178 introduced changes in the cutinase structure that are observed even in the cutinase region around the tryptophan residue. This mutation also reduces the development of hydrophobic solvent accessible patches
T179C
-
amino acid substitution, 90% of wild-type activity
T179E
-
amino acid substitution, 10% of wild-type activity
T179N
-
amino acid substitution, 119% of wild-type activity
T179Y
P00590
131% of the activity of the wild-type enzyme
T18D
-
amino acid substitution, 65% of wild-type activity
T18V
P00590
90% of the activity of the wild-type enzyme
T19V
P00590
35% of the activity of the wild-type enzyme
T45A
-
comparative structural analysis of native enzyme and mutant enzymes
T45A
P00590
98% of the activity of the wild-type enzyme
T45D
-
amino acid substitution, 54% of wild-type activity
T45K
P00590
74% of the activity of the wild-type enzyme
T50V
P00590
25% of the activity of the wild-type enzyme
T80D
P00590
32% of the activity of the wild-type enzyme
T80P
-
comparative structural analysis of native enzyme and mutant enzymes
V184A
-
activity with polyethylene terephthalate fibers is 2fold higher than wild-type enzyme, activity with polyamide 6,6 fiber is 98% of wild-type activity
W69Y
P00590
12% of the activity of the wild-type enzyme
Y119H
-
comparative structural analysis of native enzyme and mutant enzymes
Y38F
-
comparative structural analysis of native enzyme and mutant enzymes
Y38F
P00590
62% of the activity of the wild-type enzyme
H173L
Q8TGB8
36% of wild-type activity
S103A
Q8TGB8
231% of wild-type activity
S103T
Q8TGB8
38% of wild-type activity
S54D
-
site-directed mutagenesis, the mutant shows reduced transesterification activity compared to the wild-type enzyme
T179C
-
site-directed mutagenesis, the mutant shows transesterification activity similar to the wild-type enzyme, T179C displays high stability in the presence of methanol with an activity loss of only 16% as compared to 90% loss of wild-type activity, the mutant is also more stable microencapsulated in reversed micelles of bis(2-ethylhexyl) sodium sulfosuccinate in isooctane
cutinase-tryptophan,proline2
-
tryptophan tag, cutinase with varying length tryptophan tag (WP)2
cutinase-tryptophan,proline4
-
tryptophan tag, cutinase with varying length tryptophan tag (WP)4
I218A
-
engineering by site-directed mutagenesis modifying the active site, the mutant cutinase shows increased activity on polyester substrates. Mutation I218A creates space, activity on poly(ethylene terephthalate) is increased compared to the wild-type enzyme, with considerably higher hydrolysis efficiency
T101A/Q132A
-
engineering by site-directed mutagenesis modifying the active site, the mutant cutinase shows increased activity on polyester substrates. The double mutation Q132A/T101A both creates space and increases hydrophobicity. The activity of the double mutant on the soluble substrate p-nitrophenyl butyrate increased 2fold compared to wild-type cutinase, while on poly(ethylene terephthalate) the double mutant exhibits considerably higher hydrolysis efficiency
L80A
-, P41744
mutation in the hydrophobic binding loop, drastically reduces the overall activity
additional information
Q9SZW7
CDEF1-deficient mutant (SALK-014093) that carries a T-DNA insertion in the coding region of CDEF1, shows no abnormal phenotypes, such as reduced fertility or reduced lateral root emergence
additional information
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because the organism has a low but significant FAE activity, it may be easier to introduce a high level of FAE activity in cutinases through point mutations
M98C
P00590
35% of the activity of the wild-type enzyme
additional information
P00590
the insertion mutant 49aILe shows 52% of the activity of the wild-type enzyme
additional information
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a complete saturation mutagenesis approach to search cutinase for amino acids contributing to increased stability in the presence of the anionic surfactant. Mutants showing substitutions in the large hydrophobic crevice (S54D, S57D, S61D, K65P, R196A), that is thought to be the region more involved in the unfolding by anionics, will be very important to obtain an enzyme less sensitive to AOT
additional information
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stability of cutinase may be increased through mutations designed to avoid the transient formation of hydrophobic groups during protein movement. Because the organism has a low but significant ferulic acid esterase activity, it may be easier to introduce a high level of ferulic acid esterase activity in cutinases through point mutations
H204N
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is catalytically inactive. Is not covalently modified by a 4fold excess of diethyl p-nitrophenyl phosphate, in contrast to the wild-type; site-directed mutant, constructed, overexpressed, and purified
additional information
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mutant myHiC, obtained by localised random mutagenesis, shows increased activity and decreased surfactanct sensitivity
additional information
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because the organism has a low but significant FAE activity, it may be easier to introduce a high level of FAE activity in cutinases through point mutations
L153Q
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site-directed mutagenesis, the mutant shows transesterification activity similar to the wild-type enzyme
additional information
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cutinase is microencapsulated in reversed micelles of bis(2-ethylhexyl) sodium sulfosuccinate in isooctane for the production of alkyl esters, known as biodiesel, evaluation of the system stability using wild-type enzyme and three mutants, L153Q, T179C and S54D, method evaluation, overview. Loss of 45% of wild-type cutinase activity when incubated in the micellar system for 3 h, and an additional loss of 90% of the activity is observed in the presence of methanol after 10 min of incubation
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
refolding from the pH-denatured state. Existence of two distinct intermediate states in cutinase folding: an unfolding intermediate and an off-pathway folding intermediate.
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APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
degradation
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biotechnological applications of cutinases for synthetic polyester degradation
degradation
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biotechnological applications of cutinases for synthetic polyester degradation
biotechnology
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engineering new cutinase-inspired biocatalysts with tailor-made properties
degradation
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biotechnological applications of cutinases for synthetic polyester degradation
degradation
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efficient degradation of n-butyl benzyl phthalate by enzyme, degradation of 60% of initial amount within 7.5 h. Major product is 1,3-isobenzofurandione
degradation
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enzyme shows significant degradation of dipropyl phthalate to non-toxic 1,3-isobenzofurandione, with 70% degradation of initial 500 mg/l within 2.5 h
degradation
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enzyme degrades 60% of initial 500 mg/l malathion within 0.5 h, major degradation product is malathion diacid
environmental protection
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application of cutinase for degradationof dihexyl phthalate in the dihexyl phthalate-contaminated environments may be possible
industry
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useful as biocatalysts in systems involving hydrolysis, esterification, and transesterification reactions
biotechnology
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use of enzyme in a membrane reactor in presence of 1-hexanol, operational half-life of 674 days
biotechnology
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engineering new cutinase-inspired biocatalysts with tailor-made properties
biotechnology
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high-level secretion of cutinase in Pichia pastoris may be a promising alternative to many expression systems previously used for the large-scale production of cutinase in Saccharomyces cerevisiae as well as Escherichia coli. The functional expression of a large amount of extracellular cutinase offers the opportunity for developing an efficient high-throughput screening procedure for the improvement of enzymatic property and the development of novel biocatalysis of cutinase
degradation
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biotechnological applications of cutinases for synthetic polyester degradation
industry
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key for the design of biocatalysts with sufficient stability for practical applications in detergent industry
industry
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useful as biocatalysts in systems involving hydrolysis, esterification, and transesterification reactions. Cutinase proves to be the most well fitted enzyme for the detection of pesticide residues in foods even at very low levels. Use in fiber modification due to its hydrophobic nature and activity against biopolyesters present in plant cuticle. Use of cutinase to improve the wetting of cotton fibers. Cutinase is potentially useful for the removal of fats in laundry, but the unfolding of the enzyme in the presence of anionic surfactants limits its widespread use as an additive in industrial laundry detergents. Displays a stability profile that is well-fitted to the industrial process
degradation
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biotechnological applications of cutinases for synthetic polyester degradation
industry
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useful as biocatalysts in systems involving hydrolysis, esterification, and transesterification reactions; useful as biocatalysts in systems involving hydrolysis, esterification, and transesterification reactions. Displays a stability profile that is well-fitted to the industrial process
biotechnology
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immobilization of enzyme on sodium form of zeolite Y, half-life 590 days at 30C. Immobilization on zolite A, halft-life of 54 days at 30C. Half-lives after immobilization on Alumina and Accurel-PA6 are 109 and 10 days, resp. Higher temperatures induce a remarkable stability loss in all preparations. At 30C, enzymatic activities obtained wit the immobilization on zeolite A are the highest ones
biotechnology
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adsorption of enzyme onto the surface of poly(methyl methacrylate) latex particles. Up to 50% decrease in specific activity at pH-values 4.5 and 5.2. Almost no inactivation upon adsorption at pH 7.0 and 9.2. 60% increase in maximum adsorption with temperature raising from 25 to 50C
biotechnology
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study on enzyme encapsulated in sol-gel matrices prepared with alkyl-alkoxysilane precursors of different chain length. Specific activity of entrapped enzyme is comparable to enzyme immobilized on zeolite Y, with incorporation of different additives bringing about an enhancement of enzyme activity and operational stability
synthesis
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a high enzyme production, high specific enzyme activity, and high enzyme yield are obtained upon expression with a 5% air saturation of oxygen. At low dissolved oxygen concentration, enzyme yield and specific activity increase with increase of culture pH-value from 5.25 to 6.25
synthesis
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recombinant cutinase from Fusarium solani pisi is used as a catalyst in enzymatic transesterification between a mixture of triglyceride oils and methanol for biodiesel production in a bis(2-ethylhexyl) sodium sulfosuccinate (AOT)/isooctane reversed micellar system, kinetics, overview
industry
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Cutinase is used as a lipolytic enzyme in the composition of laundry and dishwashing detergents to more efficiently remove immobilized fats. The oleochemistry industries and pollutant degradation represent other potential uses of cutinase.
synthesis
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application of enginered mutant T101A/Q132A/I218A in synthetic fiber biotransformation
industry
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useful as biocatalysts in systems involving hydrolysis, esterification, and transesterification reactions. Displays a stability profile that is well-fitted to the industrial process
additional information
B9U443, -
can be exploited in treating agricultural, food, and forest raw materials as well as their processing by-products