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Literature summary for 1.8.3.2 extracted from

  • Kobayashi, J.; Sasaki, D.; Hara, K.Y.; Hasunuma, T.; Kondo, A.
    Enzymatic improvement of mitochondrial thiol oxidase Erv1 for oxidized glutathione fermentation by Saccharomyces cerevisiae (2017), Microb. Cell fact., 16, 44 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.8.3.2

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Rosetta(DE3)pLysS strain Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
D24A the mutant enzyme oxidizes GSH and gamma-glutamylcysteine at much lower rates than the wild-type enzyme Saccharomyces cerevisiae
N131A the mutant enzyme oxidizes GSH and gamma-glutamylcysteine at much lower rates than the wild-type enzyme Saccharomyces cerevisiae
N34A enhancement of catalytic activity for GSH, whereas the catalytic activity for gamma-glutamylcysteine remains unchanged. The mutant enzyme shows slightly decreased maximum temperatures at 55°C (compared to 60°C for the wild-type enzyme) Saccharomyces cerevisiae
N34Q the mutant enzyme oxidizes GSH more efficiently (201%) than the wild-type enzyme Saccharomyces cerevisiae
P129A the mutant enzyme oxidizes GSH and gamma-glutamylcysteine at much lower rates than the wild-type enzyme Saccharomyces cerevisiae
S32A the mutant enzyme oxidizes GSH more efficiently (169%) than the wild-type enzyme. About 1.5fold increase in GSSG production compared to that of the parental ERV1 gene Saccharomyces cerevisiae
S32A/N34A the mutant enzyme oxidizes GSH more efficiently (240%) than the wild-type enzyme and shows comparable activity for gamma-glutamylcysteine (96%). The mutant enzyme shows slightly decreased maximum temperatures at 55°C (compared to 60°C for the wild-type enzyme) Saccharomyces cerevisiae
S32T the mutant enzyme oxidizes GSH more efficiently (178%) than the wild-type enzyme Saccharomyces cerevisiae
S32T/N34A the mutant enzyme shows almost the same activity for GSH (192%) compared to mutant enzyme S32A, S32T, and N34A, and high activity for gamma-glutamylcysteine (161%). The mutant enzyme shows slightly decreased maximum temperatures at 55°C (compared to 60°C for the wild-type enzyme) Saccharomyces cerevisiae
W132A the mutant enzyme oxidizes GSH and gamma-glutamylcysteine at much lower rates than the wild-type enzyme Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3 3.5 glutathione pH 7.0, 30°C, mutant enzyme N34A Saccharomyces cerevisiae
4.4
-
 glutathione pH 7.0, 30°C, wild-type enzyme Saccharomyces cerevisiae
12
-
 glutathione pH 7.0, 30°C, mutant enzyme S32T Saccharomyces cerevisiae
13
-
 glutathione pH 7.0, 30°C, mutant enzyme S32A Saccharomyces cerevisiae
40.5
-
 glutathione pH 7.0, 30°C, mutant enzyme S32A/N34A Saccharomyces cerevisiae
41.2
-
 glutathione pH 7.0, 30°C, mutant enzyme S32T/N34A Saccharomyces cerevisiae

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
 Saccharomyces cerevisiae 5739
-

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-
Saccharomyces cerevisiae NBRC 10505
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 glutathione + O2
-
 Saccharomyces cerevisiae glutathione disulfide + H2O2
-
 ?
2 glutathione + O2
-
 Saccharomyces cerevisiae NBRC 10505 glutathione disulfide + H2O2
-
 ?
gamma-glutamylcysteine + O2
-
 Saccharomyces cerevisiae ?
-
 ?
gamma-glutamylcysteine + O2
-
 Saccharomyces cerevisiae NBRC 10505 ?
-
 ?

Synonyms

Synonyms Comment Organism
thiol oxidase Erv1
-
 Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
 mutant ezymes N34A, S32A/N34A and S32T/N34A Saccharomyces cerevisiae
60
-
 wild-type enzyme and mutant enzymes S32A and S32R Saccharomyces cerevisiae

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
59.1
-
 T1/2, mutant enzyme S32A/N34A Saccharomyces cerevisiae
62
-
 T1/2, mutant enzyme S32T/N34A Saccharomyces cerevisiae
64.1
-
 T1/2, mutant enzyme N34A Saccharomyces cerevisiae
68.8
-
 T1/2, mutant enzyme S32T Saccharomyces cerevisiae
69.3
-
 T1/2, mutant enzyme S32A Saccharomyces cerevisiae
90
-
 T1/2 is below 90°C, wild-type enzyme Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0002
-
 glutathione pH 7.0, 30°C, mutant enzyme S32A Saccharomyces cerevisiae
0.00083
-
 glutathione pH 7.0, 30°C, wild-type enzyme Saccharomyces cerevisiae
0.0017
-
 glutathione pH 7.0, 30°C, mutant enzyme S32T Saccharomyces cerevisiae
0.0033
-
 glutathione pH 7.0, 30°C, mutant enzyme S32T/N34A Saccharomyces cerevisiae
0.0042
-
 glutathione pH 7.0, 30°C, mutant enzyme N34A Saccharomyces cerevisiae
0.007
-
 glutathione pH 7.0, 30°C, mutant enzyme S32A/N34A Saccharomyces cerevisiae