Information on EC 3.4.22.14 - actinidain

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The expected taxonomic range for this enzyme is: Actinidia

EC NUMBER
COMMENTARY
3.4.22.14
-
RECOMMENDED NAME
GeneOntology No.
actinidain
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Similar to that of papain
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Act d 1
P00785
-
actinidain
-
-
actinidia anionic protease
-
-
-
-
actinidin
-
-
-
-
actinidin
A5HII2
-
actinidin
P00785
-
actinidin
A5HII1
-
mercaptoproteinase A2
-
-
-
-
proteinase A2 of Actinidia chinensis
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
39279-27-1
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
sarunashi or hardy kiwi
UniProt
Manually annotated by BRENDA team
cultivar Hayward
SwissProt
Manually annotated by BRENDA team
cultivar Hiward
-
-
Manually annotated by BRENDA team
var. deliciosa Hort16A and EM4
-
-
Manually annotated by BRENDA team
variety YellowA
-
-
Manually annotated by BRENDA team
cultivar Hayward
-
-
Manually annotated by BRENDA team
Lindl. var. deliciosa Hayward
-
-
Manually annotated by BRENDA team
var. deliciosa cv Hayward
-
-
Manually annotated by BRENDA team
variety Hayward
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
P00785
actinidin is important in monoallergy to kiwifruit
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-(acetamido)ethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
reactivity probe, analysis of temperature-dependence of reaction kinetics
-
-
?
2-(acetoxy)ethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
reactivity probe, analysis of temperature-dependence of reaction kinetics
-
-
?
2-(N'-acetyl-D-phenylalanyl)hydroxyethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
reactivity probe, analysis of temperature-dependence of reaction kinetics
-
-
?
2-(N'-acetyl-D-phenylalanylamino)ethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
reactivity probe, analysis of temperature-dependence of reaction kinetics
-
-
?
2-(N'-acetyl-L-phenylalanyl)hydroxyethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
reactivity probe, analysis of temperature-dependence of reaction kinetics
-
-
?
2-(N'-acetyl-L-phenylalanylamino)ethylene 2'-pyridyl disulfide + H2O
?
show the reaction diagram
-
reactivity probe, analysis of temperature-dependence of reaction kinetics
-
-
?
acetyl-Gly p-nitrophenyl ester + H2O
acetyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
acetyl-Gly-Lys methyl ester + H2O
acetyl-Gly-Lys + methanol
show the reaction diagram
-
-
-
-
?
acetyl-Lys methyl ester + H2O
acetyl-Lys + methanol
show the reaction diagram
-
-
-
-
?
benzoyl-Arg ethyl ester + H2O
benzoyl-Arg + ethanol
show the reaction diagram
-
-
-
-
?
benzoyl-Gly p-nitrophenyl ester + H2O
benzoyl-Gly + p-nitrophenol
show the reaction diagram
-
-
-
-
?
benzoyl-L-Arg ethyl ester + H2O
benzoyl-L-Arg ethanol
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Gly p-nitrophenyl ester + H2O
benzyloxycarbonyl-Gly + p-nitrophenol
show the reaction diagram
-
-
-
-
?
Benzyloxycarbonyl-Lys methyl ester + H2O
Benzyloxycarbonyl-Lys + methanol
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Lys p-nitrophenyl ester + H2O
benzyloxycarbonyl-Lys + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
butyloxycarbonyl-Phe-NH-CH2CN + H2O
?
show the reaction diagram
-
-
-
-
?
carboxybenzyl-L-Lys 4-nitrophenyl ester + H2O
carboxybenzyl-L-Lys + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
P00785
-
-
-
?
Collagen type I + H2O
?
show the reaction diagram
A5HII1, -
actinidin can hydrolyze collagen types I and II at neutral and alkaline pH
-
-
?
Collagen type I + H2O
?
show the reaction diagram
A5HII1, -
the hydrolysis of types I and II collagen by actinidin under different pH conditions is monitored by SDS-PAGE: good digestion of type I collagen at pH 7 (phosphate buffer) or pH 8.5 (Tris-HCl buffer). In citrate buffer (pH 5.5), actinidin digests more than half of the collagen type I, but in acetate buffer (pH 4), the enzyme does not hydrolyze this substrate
-
-
?
Collagen type I + H2O
?
show the reaction diagram
-
actinidain cleaves between Gly1032 and Gly1033, limited hydrolysis by actinidain protease produces monomeric collagen, which consisted almost entirely of alpha1 and alpha2 chains
-
-
?
collagen type II + H2O
?
show the reaction diagram
A5HII1, -
actinidin can hydrolyze collagen types I and II at neutral and alkaline pH
-
-
?
collagen type II + H2O
?
show the reaction diagram
A5HII1, -
the hydrolysis of types I and II collagen by actinidin under different pH conditions is monitored by SDS-PAGE: good digestion of type I collagen at pH 7 (phosphate buffer) or pH 8.5 (Tris-HCl buffer). In citrate buffer (pH 5.5), or acetate buffer (pH 4), the enzyme does not hydrolyze this substrate
-
-
?
ethyl 2-pyridyl disulfide + H2O
?
show the reaction diagram
-
reactivity probe, analysis of temperature-dependence of reaction kinetics
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
-
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
Insulin B-chain + H2O
?
show the reaction diagram
-
the enzyme cleaves most readily the carboxyl peptide bonds of those residues of the B-chain acylated by large hydrophobic residues - Val, Leu, or Phe, but not Tyr - and also the Arg22-Gly23 bond
-
-
-
N-acetyl-L-Phe-Gly methyl ester + H2O
?
show the reaction diagram
-
stopped-flow spectral analysis of increasing content of co-solvent acetonitrile on kinetics
-
-
-
N-acetyl-Phe-Gly methyl thionoester + H2O
?
show the reaction diagram
-
-
-
-
?
Nalpha-benzoyl-L-Arg p-nitroanilide + H2O
Nalpha-benzoyl-L-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
Nalpha-benzyloxycarbonyl-L-Lys p-nitrophenyl ester + H2O
Nalpha-benzyloxycarbonyl-L-Lys + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
p-toluene-sulphonyl-L-glutamine p-nitrophenyl ester + H2O
p-toluene-sulphonyl-L-glutamine + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
succinylcasein + H2O
?
show the reaction diagram
-
-
-
-
?
Tosyl-Arg methyl ester + H2O
Tosyl-Arg + methanol
show the reaction diagram
-
-
-
-
?
acid-soluble collagen + H2O
?
show the reaction diagram
-
-
-
-
?
atelocollagen + H2O
additional information
-
-
-
actinaidin cleaves the atelocollagen molecule at specific sites on the inside of the interstrand cross-linking peptides. The actinidain-processed atelocollagen shows only monomeric alpha1 and alpha2 chains, with no beta and gamma chains, atelocollagen retains its typical triple helical structure
?
kiwellin + H2O
KiTH + kissper
show the reaction diagram
-
in vitro treatment of purified kiwellin, an allergenic protein formerly isolated from green kiwi fruit, with the protease actinidin from green kiwi fruit originates KiTH, a 20 kDa protein with 100% identity with the C-terminal region of kiwellin, and kissper, a described pore-forming peptide
-
-
?
additional information
?
-
-
mechanism
-
-
-
additional information
?
-
-
the interplay of electrostatic and binding interactions determine the active center chemistry and catalytic activity
-
-
-
additional information
?
-
-
actinidain, from the juice of kiwi fruit, has no detrimental effect on either the motility of Trichuris muris or the nematode cuticle
-
-
-
additional information
?
-
A5HII1, -
actinidin compared with type II or IV collagenase isolated from intact human umbilical vein endothelial cells, hepatocytes, and thymic epithelial cells with viability more than 90%
-
-
-
additional information
?
-
-
digestibility of kiwifruit allergens, actinidin (Act d 1) and thaumatin-like protein (Act d 2), is assessed using an in vitro digestion system that approximates physiological conditions with respect to the passage of food through the stomach into the duodenum: Act d 1 precipitates in simulated gastric fluid at pH 2 and digestion of the aggregated protein proceeds slowly. The residual precipitate redissolves completely in simulated duodenal fluid at pH 6.5 and is partially digested. Act d 1 and Act d 2 display nearly unchanged IgE binding abilities
-
-
-
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
casein + H2O
?
show the reaction diagram
-
-
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
kiwellin + H2O
KiTH + kissper
show the reaction diagram
-
in vitro treatment of purified kiwellin, an allergenic protein formerly isolated from green kiwi fruit, with the protease actinidin from green kiwi fruit originates KiTH, a 20 kDa protein with 100% identity with the C-terminal region of kiwellin, and kissper, a described pore-forming peptide
-
-
?
additional information
?
-
-
actinidain, from the juice of kiwi fruit, has no detrimental effect on either the motility of Trichuris muris or the nematode cuticle
-
-
-
additional information
?
-
-
digestibility of kiwifruit allergens, actinidin (Act d 1) and thaumatin-like protein (Act d 2), is assessed using an in vitro digestion system that approximates physiological conditions with respect to the passage of food through the stomach into the duodenum: Act d 1 precipitates in simulated gastric fluid at pH 2 and digestion of the aggregated protein proceeds slowly. The residual precipitate redissolves completely in simulated duodenal fluid at pH 6.5 and is partially digested. Act d 1 and Act d 2 display nearly unchanged IgE binding abilities
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
A5HII2, -
actinidin is not dependent on cadmium
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1-(L-trans-epoxysuccinylleucylamino)-4-guanidinobutane
-
-
1-(L-trans-epoxysuccinylleucylamino)-4-guanidinobutane
-
potent, highly selective, irreversible
4,4'-dithiodipyridine
-
-
5,5'-dithiobis(2-nitrobenzoic acid)
-
-
Cystatin
-
hybrids of chicken cystatin with human kininogen domain 2 sequences exhibit improved inhibition
-
cystatin C
-
recombinant inhibitor expressed in E. coli
-
human kininogen domain 2
-
hybrids of chicken cystatin with human kininogen domain 2 sequences exhibit improved inhibition
-
KCl
-
minimal activity at 0.5-0.8 M, depending on substrate concentration. Minimal activity is 50-70% of the acitivity in absence of salt, with increase in KM-value and decrease in kcat-value. Slight reactivation above 0.8 M
L-kininogen
-
-
-
LiCl
-
minimal activity at 1.0-1.5 M, depending on substrate concentration. Minimal activity is 50-70% of the acitivity in absence of salt, with increase in KM-value and decrease in kcat-value. Slight reactivation above 1.5 M
N-acetyl-L-Arg
-
-
N-benzoyl-L-Arg
-
-
N-benzoyl-L-Arg ethyl ester
-
-
NaCl
-
minimal activity at 0.5-1.2 M, depending on substrate concentration. Minimal activity is 50-70% of the acitivity in absence of salt, with increase in KM-value and decrease in kcat-value. Slight reactivation above 1.2 M
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
dithiothreitol
-
activates
L-cysteine
-
the enzyme is activated in 20 mM potassium phosphate buffer, pH 6.5 containing 52 mM L-cysteine and 2 mM EDTA (1:1 (v/v)) for 1 h at 45C
L-cysteine
-
for activation of actinidin, 52 mM L-cysteine is added to the sample and the preparation incubated for 1 h at 45C
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
78
-
acetyl-Lys methyl ester
-
-
61
-
benzoyl-Arg ethyl ester
-
-
0.31
-
benzoyl-Gly p-nitrophenyl ester
-
-
89
-
benzoyl-L-Arg ethyl ester
-
pH 5.6, 25C, in 0.3 M KCl
21
-
benzoyl-Lys methyl ester
-
-
0.12
-
benzyloxycarbonyl-Gly p-nitrophenyl ester
-
-
0.022
-
benzyloxycarbonyl-Lys p-nitrophenyl ester
-
-
0.25
-
N-acetyl-L-Phe-Gly methyl ester
-
above 0.25 mM, at 4.3% acetonitrile, 25C, pH 5.3, parameter for pre-steady-state acylation phase
0.543
-
N-acetyl-L-Phe-Gly methyl ester
-
25C, pH 5.3, 8.3% acetonitrile parameter for pre-steady-state acylation phase
0.893
-
N-acetyl-L-Phe-Gly methyl ester
-
25C, pH 5.3, 16.7% acetonitrile, parameter for pre-steady-state acylation phase
1.8
-
Nalpha-benzoyl-L-Arg p-nitroanilide
-
pH 7.1
1.8
-
Nalpha-benzoyl-L-Arg p-nitroanilide
-
-
2.6
-
Nalpha-benzoyl-L-Arg p-nitroanilide
-
pH 4.2
11.2
-
Nalpha-benzoyl-L-Arg p-nitroanilide
-
pH 9.5
0.0262
-
Nalpha-benzyloxycarbonyl-L-Lys p-nitrophenyl ester
-
-
0.044
-
butyloxycarbonyl-Phe-NH-CH2CN
-
-
-
additional information
-
additional information
-
pH-dependence of Km-value
-
additional information
-
additional information
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.7
-
acetyl-Lys methyl ester
-
-
4.5
-
benzoyl-Arg ethyl ester
-
-
0.029
-
benzoyl-Arg p-nitroanilide
-
-
5.8
-
benzoyl-Gly p-nitrophenyl ester
-
-
2.6
-
benzoyl-L-Arg ethyl ester
-
pH 5.6, 25C, in 0.3 M KCl
3.4
-
benzyloxycarbonyl p-nitrophenyl ester
-
-
6
-
Benzyloxycarbonyl-Lys methyl ester
-
-
33
-
Nalpha-benzyloxycarbonyl-L-Lys p-nitrophenyl ester
-
-
29
-
benzyloxycarbonyl-Lys p-nitrophenyl ester
-
-
additional information
-
additional information
-
pH-dependence of turnover number
-
additional information
-
additional information
-
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.35
-
KCl
-
25C, pH 6.5
0.13
-
LiCl
-
25C, pH 6.5
0.43
-
NaCl
-
25C, pH 6.5
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.033
-
-
isoenzyme KP2
0.034
-
-
isoenzyme KP3
0.04
-
-
isoenzyme KP1
0.051
-
-
isoenzyme KP5
0.177
-
-
isoenzyme KP6
0.197
-
-
isoenzyme KP4
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
7
-
benzoyl-L-Arg ethyl ester
6
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.5
8.8
-
about 45% of maximal activity at pH 3.5 and at pH 8.8
7
8.5
A5HII1
efficient digestion of type I and type II collagen
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
A5HII1
assay at
37
-
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
actinidin mRNAs for acidic and basic proteins are expressed at comparable levels throughout ripening. Actinidin mRNA expression is highest in fruit at harvest, expression decreases as fruit ripened and is much lower in the core compared with outer pericarp tissue. Low levels of a basic actinidin protein (KFAct2a) in ripe Actinidia chinensis fruit. Extremely high levels of an acidic actinidin protein (KFAct1a) is detected in Actinidia chinensis EM4 fruit. This acidic protein appears to be absent in Actinidia chinensis Hort16A
Manually annotated by BRENDA team
-
actinidin mRNAs for acidic and basic proteins are expressed at comparable levels throughout ripening. Actinidin mRNA expression is highest in fruit at harvest, expression decreases as fruit ripened and is much lower in the core compared with outer pericarp tissue. Low levels of a basic actinidin protein (KFAct2a) in ripe Actinidia deliciosa fruit. Extremely high levels of an acidic actinidin protein (KFAct1a) is detected in Actinidia deliciosa fruit. Both the basic and acidic actinidin isoforms in Actinidia deliciosa are active cysteine proteases
Manually annotated by BRENDA team
-
compared to Act d 8 and Act c 8, Act d 1 transcript is stronger expressed as determined by RT-PCR
Manually annotated by BRENDA team
-
actinidin is present in small cells, but not large cells in the outer pericarp of mature Actinidia deliciosa fruit at harvest. Within the small cells, actinidin is localised diffusely in the vacuole, associated with the plasma membrane, and in a layer in the plastids near starch granules
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
actinidin is present in small cells, but not large cells in the outer pericarp of mature Actinidia deliciosa fruit at harvest. Within the small cells, actinidin is localised diffusely in the vacuole, associated with the plasma membrane, and in a layer in the plastids near starch granules
Manually annotated by BRENDA team
-
actinidin is present in small cells, but not large cells in the outer pericarp of mature Actinidia deliciosa fruit at harvest. Within the small cells, actinidin is localised diffusely in the vacuole, associated with the plasma membrane, and in a layer in the plastids near starch granules
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
12800
-
-
gel filtration
23000
-
-
SDS-PAGE
26000
-
-
high speed equilibrium method
27000
-
P00785
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
A5HII2, -
x * 24000, SDS-PAGE
?
-
x * 22000, active enzyme, SDS-PAGE; x * 30000, thermally inactivated enzyme, SDS-PAGE
?
-
x * 23500, SDS-PAGE
?
-
x * 23500, calculation from amino acid composition
?
-
x * 27000, SDS-PAGE
?
-
x * 27445, calculation from amino acid sequence; x * 30000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapor diffusion method, using 30 mM CdCl2 and 30% (w/v) PEG 400 in 0.1 M sodium acetate pH 4.6, at 15C
A5HII2, -
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.1
-
-
1-2 s, 40% of the catalytic sites survive
2.4
-
-
1-2 s, 75% of the catalytic sites survive
2.5
-
-
above, 1-2 s, stable
additional information
-
-
thermal stability of Act d 1 and Act d 2 is strongly pH dependent. Act d 1 is irreversibly destabilized in acidic solutions (pH 3.5)
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
95
-
-
the enzyme is thermally inactivated after 5 min at 95C
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acetonitrile
-
stopped-flow spectral analysis of increasing content of co-solvent acetonitrile on kinetics. With low acetonitrile content, plots of reaction are linear, increasing the acetonitrile content results in marked deviations of the plots from linearity with a rate minimum around [S]0 of 0.15 mM
dimethyl sulfoxide
-
comparison of effect on reaction kinetics with acetonitrile
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
inactivated by methylene blue-catalyzed photooxidation, at pH 7.9 and 20C. The rate of inactivation is pH-dependent and becomes slower at lower pH values
-
81476
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, crystalline enzyme, resuspended in dialysis buffer at pH 4, stable for several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
actinidin can be separated into six proteases, named KP1, KP2, KP3, KP4, KP5 and KP6
-
ammonium sulfate precipitation and DEAE-Sephadex gel filtration
-
multiple forms
-
; the enzyme fraction is precipitated from kiwifruit extract by 60% saturation of ammonium sulfate. The precipitate is redissolved in 50 mM citrate buffer (pH 5.5) and dialyzed overnight against this buffer. The dialyzate is loaded into a DEAE-Sepharose Fast Flow column, which pre-equilibrated with the same buffer. The adsorbed fractions are eluted with 0.0-1 M linear gradient of sodium chloride in the buffer
A5HII1
Act d 1 is purified from kiwifruit extracts by covalent chromatography using activated thiopropyl sepharose 6B
-
Actinidin is purified from the soluble fraction of kiwi fruit by ion exchange chromatography on DE-52 and Mono-Q columns
-
DEAE-Sephadex A-50 gel filtration and SP-Sephadex C-50 gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Nicotiana benthamiana T1, T2, T3, and T4 transgenic lines
-
expression in Escherichia coli as inclusion bodies
-
expressed in Nicotiana benthamiana T1, T2, T3, and T4 transgenic lines
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
P75Q
-
the mutant shows reduced activity compared tot he wild type enzyme
proteolytic modification
-
the enzyme is secreted in an inactive form as zymogen, named actininidin, which has a MW of 39000 Da
S68G
-
inactive
S68G/P75Q
-
the mutant shows reduced activity compared tot he wild type enzyme
P75Q
-
the mutant shows reduced activity compared tot he wild type enzyme
S68G
-
inactive
S68G/P75Q
-
the mutant shows reduced activity compared tot he wild type enzyme
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
food industry
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actinidin is used as a beef tenderizer, use of actinidin-tenderized beef significantly improves emulsion stability, texture, and organoleptic properties of the sausage product
medicine
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to isolate major kiwifruit allergens a large group of kiwi-sensitized patients with different clinical symptoms are investigated. The three major IgE-binding proteins from kiwifruit extracts are isolated and characterized. The isolated kiwifruit allergens are: actinidin Act d 1, glycosylated thaumatin-like Act d 2 and a novel 40 kDa glycoprotein designated as Act d 3.02. Specific IgE to each of the three allergens is found in over 60% of sera from kiwi-sensitized patients, and Act d 1 and Act d 2 induces positive SPT responses in over 50% of the tested patients. A significant link between IgE levels to Act d 1 and Act d 3 and anaphylaxis is uncovered. Act d 1, Act d 2 and Act d 3 are major allergens in the population studied. Severe symptoms after kiwi ingestion are associated with high IgE levels to Act d 1 and Act d 3
medicine
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actinidin is a biomarker of kiwifruit allergy
molecular biology
A5HII1
actinidin compared with type II or IV collagenase isolates intact human umbilical vein endothelial cells, hepatocytes, and thymic epithelial cells with viability more than 90%
nutrition
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actinidin induces protease-dependent morphology changes of T84 human colorectal adenocarcinoma cells leading to cell rounding and desquamation of the epithelial monolayer, without affecting cell viability
synthesis
A5HII1
the enzyme can be used efficiently for hydrolysis of collagen and isolation of different cell populations from various solid tissues