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Literature summary for 4.2.2.2 extracted from

  • Zhou, C.; Ye, J.; Xue, Y.; Ma, Y.
    Directed evolution and structural analysis of alkaline pectate lyase from the alkaliphilic bacterium Bacillus sp. strain N16-5 to improve its thermostability for efficient ramie degumming (2015), Appl. Environ. Microbiol., 81, 5714-5723 .
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
E124I mutant displays increased thermostability Bacillus sp. N16-5
E124I/T178A/N186D/I211V/A254T/S271G/N289H 140fold increase in the t50 value at 50°C, accompanied by an 84.3% decrease in activity Bacillus sp. N16-5
K313R mutation does not improve thermostability Bacillus sp. N16-5
N186D/I211V/A254T/S271G/N289H 24fold increase in the t50 value at 50°C, with a 23.3% increase in activity Bacillus sp. N16-5
R72S mutation does not improve thermostability Bacillus sp. N16-5
S233C mutation does not improve thermostability Bacillus sp. N16-5
S271G mutant displays increased thermostability Bacillus sp. N16-5
S308L mutation does not improve thermostability Bacillus sp. N16-5
T178A mutant displays increased thermostability Bacillus sp. N16-5

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
polygalacturonic acid Km value is 1.13 mg/ml for wild-type, 1.20 mg/ml for mutant S271G, 1.12 mg/ml for mutant E241I, 1.28 mg/ml for mutant T178A, 1.74 mg/ml for mutant N186D/I211V/A254T/S271G/N289H, and 3.96 mg/ml for mutant 124I/T178A/N186D/I211V/A254T/S271G/N289H, respectively, pH 10.5, 50°C Bacillus sp. N16-5

Organism

Organism UniProt Comment Textmining
Bacillus sp. N16-5 D0VP31
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
293
-
mutant 124I/T178A/N186D/I211V/A254T/S271G/N289H, pH 10.5, 50°C Bacillus sp. N16-5
1870
-
wild-type, pH 10.5, 50°C Bacillus sp. N16-5
2305
-
mutant N186D/I211V/A254T/S271G/N289H, pH 10.5, 50°C Bacillus sp. N16-5

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
polygalacturonic acid
-
Bacillus sp. N16-5 unsaturated oligogalacturonides
-
?
ramie fiber
-
Bacillus sp. N16-5 ?
-
?

Synonyms

Synonyms Comment Organism
PelA
-
Bacillus sp. N16-5

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
time at which the enzyme loses 50% of its initial activity, i.e. t50 value, is 10 min for wild-type, 240 min for mutant N186D/I211V/A254T/S271G/N289H, 1400 min for mutant E124I/T178A/N186D/I211V/A254T/S271G/N289H, respectively Bacillus sp. N16-5

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
526
-
polygalacturonic acid mutant 124I/T178A/N186D/I211V/A254T/S271G/N289H, pH 10.5, 50°C Bacillus sp. N16-5
1532
-
polygalacturonic acid mutant T178A, pH 10.5, 50°C Bacillus sp. N16-5
2492
-
polygalacturonic acid mutant E241I, pH 10.5, 50°C Bacillus sp. N16-5
3319
-
polygalacturonic acid wild-type, pH 10.5, 50°C Bacillus sp. N16-5
3637
-
polygalacturonic acid mutant S271G, pH 10.5, 50°C Bacillus sp. N16-5
3932
-
polygalacturonic acid mutant N186D/I211V/A254T/S271G/N289H, pH 10.5, 50°C Bacillus sp. N16-5

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
additional information
-
polygalacturonic acid kcat/Km value is 2932 ml/mg/s for wild-type, 3044 ml/mg/s for mutant S271G, 2219 ml/mg/s for mutant E241I, 1196 ml/mg/s for mutant T178A, 2255 ml/mg/s for mutant N186D/I211V/A254T/S271G/N289H, and 133 ml/mg/s for mutant 124I/T178A/N186D/I211V/A254T/S271G/N289H, respectively, pH 10.5, 50°C Bacillus sp. N16-5