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Literature summary extracted from

  • Li, G.; Fang, X.; Su, F.; Chen, Y.; Xu, L.; Yan, Y.
    Enhancing the thermostability of Rhizomucor miehei lipase with a limited screening library by rational-design point mutations and disulfide bonds (2018), Appl. Environ. Microbiol., 84, e02129-17 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.3 expression in Escherichia coli Rhizomucor miehei

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.3 T18K/T22I/E230I thermostable mutant, melting temperature is 10 degrees higher than wild-type Rhizomucor miehei
3.1.1.3 T18K/T22I/S56C/N63C/V189C/E230I/D238C thermostable mutant, residues S56C/N63C and V189C/D238C may form additional disulfide bonds. Melting temperature is 14.3 degrees higher than wild-type and half-life at 70°C is 12.5fold increased Rhizomucor miehei

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.3 1.17
-
 4-nitrophenyl octanoate wild-type, pH 8.0, 45°C Rhizomucor miehei
3.1.1.3 1.53
-
 4-nitrophenyl octanoate mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, pH 8.0, 60°C Rhizomucor miehei
3.1.1.3 2.01
-
 4-nitrophenyl octanoate mutant T18K/T22I/E230I, pH 8.0, 55°C Rhizomucor miehei

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.3 Rhizomucor miehei A0A2D3HJ61
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.3 4-nitrophenyl octanoate + H2O
-
 Rhizomucor miehei 4-nitrophenol + octanoate
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.3 45
-
 wild-type Rhizomucor miehei
3.1.1.3 55
-
 mutant T18K/T22I/E230I Rhizomucor miehei
3.1.1.3 60
-
 mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C Rhizomucor miehei

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.3 58.7
-
 melting temperature, wild-type Rhizomucor miehei
3.1.1.3 65
-
 120 min, 13% residual activity for wild-type, 95% for mutant T18K/T22I/E230I Rhizomucor miehei
3.1.1.3 68.1
-
 melting temperature, mutant T18K/T22I/E230I Rhizomucor miehei
3.1.1.3 70
-
 wild-type, half-life 2.2 min, mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, half-life 27.5 min Rhizomucor miehei
3.1.1.3 73
-
 melting temperature, mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C Rhizomucor miehei

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.3 24.05
-
 4-nitrophenyl octanoate wild-type, pH 8.0, 45°C Rhizomucor miehei
3.1.1.3 42.34
-
 4-nitrophenyl octanoate mutant T18K/T22I/E230I, pH 8.0, 55°C Rhizomucor miehei
3.1.1.3 43.71
-
 4-nitrophenyl octanoate mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, pH 8.0, 60°C Rhizomucor miehei

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.3 8
-
 wild-type Rhizomucor miehei
3.1.1.3 8.5
-
 mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C Rhizomucor miehei

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.1.3 20.6
-
 4-nitrophenyl octanoate wild-type, pH 8.0, 45°C Rhizomucor miehei
3.1.1.3 21.1
-
 4-nitrophenyl octanoate mutant T18K/T22I/E230I, pH 8.0, 55°C Rhizomucor miehei
3.1.1.3 28.6
-
 4-nitrophenyl octanoate mutant T18K/T22I/S56C/N63C/V189C/E230I/D238C, pH 8.0, 60°C Rhizomucor miehei