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Literature summary for 4.4.1.13 extracted from

  • Holt, S.; Cordente, A.G.; Williams, S.J.; Capone, D.L.; Jitjaroen, W.; Menz, I.R., Curtin, C.; Anderson, P.A.
    Engineering Saccharomyces cerevisiae to release 3-mercaptohexan-1-ol during fermentation through overexpression of an S. cerevisiae gene, STR3, for improvement of wine aroma (2011), Appl. Environ. Microbiol., 77, 3626-3632.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.4.1.13

Application

Application Comment Organism
food industry improvement of wine aroma during fermentation of a Vitis vinifera cultivar Sauvignon blanc juice Saccharomyces cerevisiae

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Saccharomyces cerevisiae strain VIN 13 and in Escherichia coli BL21(DE3) cells Saccharomyces cerevisiae

General Stability

General Stability Organism
the buffer exchange conditions to stabilize the isolated protein and maintain its enzymatic activity include 500 mM KCl and 20 mM EDTA. By including 10% (w/v) glycerol in the buffer, less than 6% of the activity is lost in a freeze-thaw cycle, compared to the 56% loss in the presence of 10 mM imidazole Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.096
-
 L-cystathionine in 50 mM phosphate buffer, pH 8.5, 0.02 mM pyridoxal 5'-phosphate, 1 mM EDTA, at 37°C Saccharomyces cerevisiae
0.178
-
 L-djenkolate in 50 mM phosphate buffer, pH 8.5, 0.02 mM pyridoxal 5'-phosphate, 1 mM EDTA, at 37°C Saccharomyces cerevisiae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
52000
-
 4 * 52000, SDS-PAGE Saccharomyces cerevisiae
53000
-
 4 * 53000, estimated from amino acid sequence Saccharomyces cerevisiae
195000
-
 gel filtration Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-cystathionine + H2O Saccharomyces cerevisiae 100% activity L-homocysteine + NH3 + pyruvate
-
 ?
L-cystathionine + H2O Saccharomyces cerevisiae EC1118 100% activity L-homocysteine + NH3 + pyruvate
-
 ?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae E3VL26
-
-
Saccharomyces cerevisiae EC1118 E3VL26
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-Sepharose column chromatography Saccharomyces cerevisiae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1258
-
 using L-cystathionine as substrate,in 50 mM phosphate buffer, pH 8.5, 0.02 mM pyridoxal 5'-phosphate, 1 mM EDTA, at 37°C Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(S)-4-(4-methylpentan-2-one)-L-cysteine + H2O
-
 Saccharomyces cerevisiae 4-mercapto-4-methylpentan-2-one + NH3 + pyruvate Str3p is able to release 0.0123 mM 4-mercapto-4-methylpentan-2-one from 2 mM concentration of its precursor S-(2-methyl-4-oxopentan-2-yl)-L-cysteine ?
L-cystathionine + H2O 100% activity Saccharomyces cerevisiae L-homocysteine + NH3 + pyruvate
-
 ?
L-cystathionine + H2O 100% activity Saccharomyces cerevisiae EC1118 L-homocysteine + NH3 + pyruvate
-
 ?
L-cysteine + H2O 1.0% activity compared to L-cystathionine Saccharomyces cerevisiae ?
-
 ?
L-cysteine + H2O 1.0% activity compared to L-cystathionine Saccharomyces cerevisiae EC1118 ?
-
 ?
L-cystine + H2O 22% activity compared to L-cystathionine Saccharomyces cerevisiae ?
-
 ?
L-djenkolate + H2O most effective substrate, 154% activity compared to L-cystathionine Saccharomyces cerevisiae pyruvate + NH3 + ?
-
 ?
additional information no alpha-ketobutyrate is detected with any of the substrates susceptible for beta-lyase activity, confirming that Str3p has only beta-lyase activity. L-methionine is no substrate for Str3p Saccharomyces cerevisiae ?
-
 ?
additional information no alpha-ketobutyrate is detected with any of the substrates susceptible for beta-lyase activity, confirming that Str3p has only beta-lyase activity. L-methionine is no substrate for Str3p Saccharomyces cerevisiae EC1118 ?
-
 ?
S-(1-hydroxyhexan-3-yl)-L-cysteine + H2O
-
 Saccharomyces cerevisiae 3-mercaptohexan-1-ol + NH3 + pyruvate Str3p is able to release 0.0021 mM 3-mercaptohexan-1-ol from 2 mM concentrations of its precursor (S)-3-(hexan-1-ol)-L-cysteine ?
S-ethyl-L-cysteine 9.0% activity compared to L-cystathionine Saccharomyces cerevisiae ?
-
 ?
S-ethyl-L-cysteine 9.0% activity compared to L-cystathionine Saccharomyces cerevisiae EC1118 ?
-
 ?
S-methyl-L-cysteine + H2O 7.4% activity compared to L-cystathionine Saccharomyces cerevisiae ?
-
 ?
S-methyl-L-cysteine + H2O 7.4% activity compared to L-cystathionine Saccharomyces cerevisiae EC1118 ?
-
 ?

Subunits

Subunits Comment Organism
homotetramer 4 * 52000, SDS-PAGE Saccharomyces cerevisiae
homotetramer 4 * 53000, estimated from amino acid sequence Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
CBL
-
 Saccharomyces cerevisiae
Str3p
-
 Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.91
-
 L-cystathionine in 50 mM phosphate buffer, pH 8.5, 0.02 mM pyridoxal 5'-phosphate, 1 mM EDTA, at 37°C Saccharomyces cerevisiae
1.27
-
 L-djenkolate in 50 mM phosphate buffer, pH 8.5, 0.02 mM pyridoxal 5'-phosphate, 1 mM EDTA, at 37°C Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.8
-
 using L-cystathionine as substrate Saccharomyces cerevisiae

pH Range

pH Minimum pH Maximum Comment Organism
7 10.5 using L-cystathionine as substrate, no significant activity is observed below pH 7.0 or above pH 10.5 Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate dependent on Saccharomyces cerevisiae

Expression

Organism Comment Expression
Saccharomyces cerevisiae STR3 transcript levels in the modified strain VIN 13 are 18fold higher at day 5 but only 50% higher by the end of grape juice fermentation. In the VIN 13 control strain, the level of STR3 transcript under the control of its native promoter increase 12.3fold between day 5 and day 15 of fermentation up