Any feedback?
Please rate this page
(enzyme.php)
(0/150)

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 1.10.3.2 - laccase and Organism(s) Thermus thermophilus and UniProt Accession Q72HW2

for references in articles please use BRENDA:EC1.10.3.2
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
     1 Oxidoreductases
         1.10 Acting on diphenols and related substances as donors
             1.10.3 With oxygen as acceptor
                1.10.3.2 laccase
IUBMB Comments
A group of multi-copper proteins of low specificity acting on both o- and p-quinols, and often acting also on aminophenols and phenylenediamine. The semiquinone may react further either enzymically or non-enzymically.
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Thermus thermophilus
UNIPROT: Q72HW2
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
The taxonomic range for the selected organisms is: Thermus thermophilus
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Reaction Schemes
Synonyms
laccase, lacc, phenol oxidase, laccase a, cota-laccase, lac i, poxa1b, diphenol oxidase, laccase2, cota laccase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Benzenediol:oxygen oxidoreductase
Diphenol oxidase
-
-
-
-
Laccase allele OR
-
-
-
-
Laccase allele TS
-
-
-
-
Ligninolytic phenoloxidase
-
-
-
-
p-diphenol oxidase
-
-
-
-
urishiol oxidase
-
-
-
-
urushiol oxidase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
benzenediol:oxygen oxidoreductase
A group of multi-copper proteins of low specificity acting on both o- and p-quinols, and often acting also on aminophenols and phenylenediamine. The semiquinone may react further either enzymically or non-enzymically.
CAS REGISTRY NUMBER
COMMENTARY hide
80498-15-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonate) + O2
?
show the reaction diagram
-
-
-
?
2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate) + O2
?
show the reaction diagram
i.e. ABTS
-
-
?
2,6-dimethoxyphenol + O2
?
show the reaction diagram
-
-
-
?
4-fluoro-2-methylphenol + O2
?
show the reaction diagram
-
-
-
?
guaiacol + O2
?
show the reaction diagram
-
-
-
?
syringaldazine + O2
?
show the reaction diagram
-
-
-
?
Congo Red + O2
?
show the reaction diagram
the enzyme decolorizes the azo dye
-
-
?
guaiacol + O2
6-methoxycyclohexa-2,4-dienone + H2O
show the reaction diagram
-
-
-
?
guaiacol + O2
?
show the reaction diagram
-
-
-
-
?
Reactive Black B + O2
?
show the reaction diagram
the enzyme decolorizes the azo dye
-
-
?
Reactive Black WNN + O2
?
show the reaction diagram
the enzyme decolorizes the azo dye
-
-
?
Remazol Brilliant Blue R + O2
?
show the reaction diagram
the enzyme decolorizes the anthraquinone dye
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
the enzyme oxidizes a range of phenolic and nonphenolic compounds using molecular oxygen as an electron acceptor to produce water with a broad substrate specificity
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cu2+
the enzyme requires copper ions for activity. The dependence of the activity on Cu2+ concentration is sigmoidal with the midpoint at 0.00304 mM. Other metal ions (Mg2+, Mn2+, Ca2+, Ni2+, or Zn2+ each at 1 mM) fail to support the activity. The methionine-rich region from Met271 to Met283 may be involved in copper binding
copper
multicopper oxidase
Cu2+
-
a multicopper protein
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
the reaction in the presence of 0.1 mM CuSO4 is completely inhibited by the addition of 1 mM EDTA
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.9 - 4.4
2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonate)
0.31 - 2.94
2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate)
0.0153 - 1.88
syringaldazine
0.215 - 0.575
guaiacol
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.58 - 24.6
2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonate)
0.15 - 0.69
2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate)
3.66 - 10.3
syringaldazine
0.8 - 8.4
guaiacol
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.065 - 0.521
2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonate)
3.3 - 22
guaiacol
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 5.5
maximal activity for 2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonate) and syringaldazine
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60
reaction time: 180 min
90
reactions time: less than 1 h
92
at pH 5.0, maximal activity for 2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonate)
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40 - 85
40°C: about 40% of maximal activity, 85°C: about 55% of maximal activity
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
48986
calculated from sequence
53000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
1 * 5300, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
by signal peptide prediction, the enzyme is assumed to be a secretory protein starting from Gln23
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
two-domain-type by sitting-drop vapour-diffusion method, mixing of 0.001 ml of a protein solution, containing 8.5 mg/ml protein, with 0.001 ml of reservoir solution, containing 0.2 M potassium chloride, 0.05 M HEPES, pH 7.5, and 35% v/v pentaerythritol propoxylate, at 20°C, X-ray diffraction structure determination and analysis at 1.7 A resolution, single-wavelength anomalous diffraction technique using Cu atoms
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
M455L
mutation in T1 Cu site, incorporation of 3-4 copper atoms, similar to wild-type. Mutation results in an increase of 100 mV in the O2 reduction potential, while the enzymatic activity for ABTS oxidation is decreased
M456A
mutation in T1 Cu site, incorporation of 3-4 copper atoms, similar to wild-type
D394E
mutant with the lower laccase activity displays a decreased decolorization efficiency as compared to the wild-type enzyme. Expressed in a lower level, about 50%, of the wild type enzyme. Optimum pH shifts towards the acidic value (0.5-1 units) relative to the wild type enzyme which has an optimal pH 6.0
D394M
mutant with the lower laccase activity displays a decreased decolorization efficiency as compared to the wild-type enzyme. Expressed in a lower level, about 50%, of the wild type enzyme. Optimum pH shifts towards the acidic value (0.5-1 units) relative to the wild type enzyme which has an optimal pH 6.0
D394R
mutant with the lower laccase activity displays a decreased decolorization efficiency as compared to the wild-type enzyme. Expressed in a lower level, about 16%, of the wild type enzyme. Optimum pH shifts towards the acidic value (0.5-1 units) relative to the wild type enzyme which has an optimal pH 6.0
D396A
mutant enzyme with higher catalytic efficiency decolorizes the synthetic dye more efficiently than the wild-type enzyme
D396E
mutant enzyme with higher catalytic efficiency decolorizes the synthetic dye more efficiently than the wild-type enzyme
D396M
mutant enzyme with higher catalytic efficiency decolorizes the synthetic dye more efficiently than the wild-type enzyme
K428E
-
1.3fold decrease in kcat/Km for the substrate guaiacol
K428L
-
1.6fold decrease in kcat/Km for the substrate guaiacol. 70% decrease in activity of mutant enzyme after 4 h at 80°C. 30% decrease in activity of wild-type enzyme after 4 h at 80°C
K428M
-
1.4fold increase in kcat/Km for the substrate guaiacol
K428R
-
1.3fold decrease in kcat/Km for the substrate guaiacol
additional information
-
construction of a two-type domain laccase from a three-type domain laccase by eliminating the signal sequence domain
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
the enzyme undergoes slow partial unfolding and thermal inactivation above 65°C, making it unsuitable for long incubations above this temperature
80
half-life: more than 14 h
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged two-type domain laccase from Escherichia coli strain BL21(DE3) by heat treatment, centrifugation, nickel affinity chromatography, and ultrafiltration
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
by signal peptide prediction, the enzyme is assumed to be a secretory protein starting from Gln23. The DNA encoding the mature protein is then cloned and expressed in Escherichia coli BL21 (DE3). The recombinant enzyme, expressed as an apoprotein, is dialyzed against copper-containing buffer to yield a holoprotein
overexpression in Escherichia coli
expression in Escherichia coli BL21
-
expression in Escherichia coli BL21(DE3)
expression of a His-tagged two-type domain laccase, lacking the signal sequence domain, in Escherichia coli strain BL21(DE3)
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
copper-inducible enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
industry
the enzyme decoloures up to six different industrial dyes, with or without the use of redox mediators such as ABTS
additional information
-
the enzyme may be used in a variety of biotechnological applications, including textile dye bleaching, pulp bleaching, bioremediation, polymer synthesis and biosensors
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Komori, H.; Miyazaki, K.; Higuchi, Y.
Crystallization and preliminary X-ray diffraction analysis of a putative two-domain-type laccase from a metagenome
Acta Crystallogr. Sect. F
65
264-266
2009
Thermus thermophilus
Manually annotated by BRENDA team
Kumari, A.; Kishor, N.; Guptasarma, P.
Characterization of a mildly alkalophilic and thermostable recombinant Thermus thermophilus laccase with applications in decolourization of dyes
Biotechnol. Lett.
40
285-295
2018
Thermus thermophilus (Q72HW2), Thermus thermophilus
Manually annotated by BRENDA team
Miyazaki, K.
A hyperthermophilic laccase from Thermus thermophilus HB27
Extremophiles
9
415-425
2005
Thermus thermophilus (Q72HW2)
Manually annotated by BRENDA team
Clement, R.; Wang, X.; Biaso, F.; Ilbert, M.; Mazurenko, I.; Lojou, E.
Mutations in the coordination spheres of T1 Cu affect Cu2+-activation of the laccase from Thermus thermophilus
Biochimie
182
228-237
2021
Thermus thermophilus (Q72HW2), Thermus thermophilus
Manually annotated by BRENDA team
Zhang, Y.; Dai, Z.; Zhang, S.; Yang, X.
The catalytic properties of Thermus thermophilus SG0.5JP17-16 laccase were regulated by the conformational dynamics of pocket loop 6
Biochim. Biophys. Acta Gen. Subj.
1865
129872
2021
Thermus thermophilus (F6DF14), Thermus thermophilus SG0.5JP17-16 (F6DF14), Thermus thermophilus SG0.5JP17-16
Manually annotated by BRENDA team
Zhu, Y.; Zhan, J.; Zhang, Y.; Lin, Y.; Yang, X.
The K428 residue from Thermus thermophilus SG0.5JP17-16 laccase plays the substantial role in substrate binding and oxidation
J. Biomol. Struct. Dyn.
39
1312-1320
2021
Thermus thermophilus, Thermus thermophilus SG0.5JP17-16
Manually annotated by BRENDA team