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Literature summary for 3.5.1.11 extracted from

  • Xu, G.; Zhao, Q.; Huang, B.; Zhou, J.; Cao, F.
    Directed evolution of a penicillin V acylase from Bacillus sphaericus to improve its catalytic efficiency for 6-APA production (2018), Enzyme Microb. Technol., 119, 65-70 .
    View publication on PubMed
Search for more literature for 3.5.1.11

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21 (DE3) Lysinibacillus sphaericus

Protein Variants

Protein Variants Comment Organism
N198Y the mutant enzyme shows 1.36fold higher specific activity compared to wild-type enzyme Lysinibacillus sphaericus
N198Y/S110C the mutant enzyme shows 2.26fold higher specific activity compared to wild-type enzyme Lysinibacillus sphaericus
T63S the mutant enzyme shows 11.14fold higher specific activity compared to wild-type enzyme Lysinibacillus sphaericus
T63S/N198Y the mutant enzyme shows 4.6fold higher specific activity compared to wild-type enzyme Lysinibacillus sphaericus
T63S/S110C the mutant enzyme shows 3.97fold higher specific activity compared to wild-type enzyme Lysinibacillus sphaericus
T63S/S110C/N198Y the mutant enzyme shows 12.4fold higher specific activity and 11.3fold higher catalytic efficiency compared to wild-type enzyme. The mutant enzyme has a potential for large-scale industrial application for 6-aminopenicillanic acid production Lysinibacillus sphaericus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
21.95
-
 Penicillin V pH 6.0, 37°C, wild-type enzyme Lysinibacillus sphaericus
26.39
-
 Penicillin V pH 6.0, 37°C, mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
 SDS-PAGE Lysinibacillus sphaericus

Organism

Organism UniProt Comment Textmining
Lysinibacillus sphaericus P12256
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Lysinibacillus sphaericus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.9
-
 pH 6.0, 37°C, wild-type enzyme Lysinibacillus sphaericus
7.85
-
 pH 6.0, 37°C, mutant enzyme S110C Lysinibacillus sphaericus
9.37
-
 pH 6.0, 37°C, mutant enzyme N198Y Lysinibacillus sphaericus
11.87
-
 pH 6.0, 37°C, mutant enzyme T63S Lysinibacillus sphaericus
15.59
-
 pH 6.0, 37°C, mutant enzyme N198Y/S110C Lysinibacillus sphaericus
27.41
-
 pH 6.0, 37°C, mutant enzyme T63S/S110C Lysinibacillus sphaericus
31.72
-
 pH 6.0, 37°C, mutant enzyme T63S/N198Y Lysinibacillus sphaericus
92.49
-
 pH 6.0, 37°C, mutant enzyme T63S/N198Y/S110C Lysinibacillus sphaericus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
penicillin V + H2O the conversion yields of 6-aminopenicillanate catalyzed by mutant enzyme T63S/S110C/N198Y reaches 98% with 20% (w/v) penicillin V as substrate, which is significantly higher than that of the wild-type enzyme (85%). The enhancement of specific activity of mutant enzyme is probably attributed to the changes in the number of hydrogen bonds within the molecules Lysinibacillus sphaericus 6-aminopenicillanate + phenoxyacetate
-
 ?

Subunits

Subunits Comment Organism
? x * 35000, SDS-PAGE Lysinibacillus sphaericus

Synonyms

Synonyms Comment Organism
penicillin V acylase
-
 Lysinibacillus sphaericus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
 mutant enzyme T63S/N198Y/S110C Lysinibacillus sphaericus
60
-
 wild-type enzyme Lysinibacillus sphaericus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 60 40°C: about 40% of maximal activity, 60°C: about 40% of maximal activity, mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus
40 65 40°C: about 40% of maximal activity, 65°C: about 50% of maximal activity, wild-type enzyme Lysinibacillus sphaericus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
 pH 6.0, 120 min, less than 5% loss of activity, wild-type enzyme and mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus
40
-
 pH 6.0, 120 min, 5% loss of activity, wild-type enzyme and mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus
50
-
 pH 6.0, 120 min, about 75% loss of activity, wild-type enzyme. pH 6.0, 120 min, about 85% loss of activity, mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus
60
-
 pH 6.0, 120 min, about 75% loss of activity, wild-type enzyme. pH 6.0, 120 min, about 95% loss of activity, mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.38
-
 Penicillin V pH 6.0, 37°C, wild-type enzyme Lysinibacillus sphaericus
94.59
-
 Penicillin V pH 6.0, 37°C, mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 wild-type enzyme and mutant enzyme T63S/N198Y/S110C Lysinibacillus sphaericus

pH Range

pH Minimum pH Maximum Comment Organism
5.5 6.5 pH 5.5: about 65% of maximal activity, pH 6.5: about 60% of maximal activity, mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus
5.5 6.5 pH 5.5: about 70% of maximal activity, pH 6.5: about 75% of maximal activity, wild-type enzyme Lysinibacillus sphaericus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.29
-
 Penicillin V pH 6.0, 37°C, wild-type enzyme Lysinibacillus sphaericus
3.58
-
 Penicillin V pH 6.0, 37°C, mutant enzyme T63S/S110C/N198Y Lysinibacillus sphaericus