Improvement of the acid stability of Bacillus licheniformis alpha amylase by site-directed mutagenesis

2017 ◽  
Vol 58 ◽  
pp. 174-180 ◽  
Author(s):  
Yihan Liu ◽  
Lin Huang ◽  
Leibo Jia ◽  
Shuang Gui ◽  
Yu Fu ◽  
...  
Keyword(s):  
Bacillus Licheniformis ◽  
Site Directed Mutagenesis ◽  
Alpha Amylase ◽  
Directed Mutagenesis ◽  
Acid Stability

Enhancement of Acid Stability of Alpha Amylase from Bacillus licheniformis by Error-Prone PCR

2013 ◽  
Vol 774-776 ◽  
pp. 664-669
Author(s):  
Yan Jing Xu ◽  
Yi Han Liu ◽  
Shuai Fan ◽  
Fu Ping Lu
Keyword(s):  
Bacillus Licheniformis ◽  
Double Mutant ◽  
Kinetic Studies ◽  
Site Directed Mutagenesis ◽  
Alpha Amylase ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
Chain Reaction ◽  
Acid Stability ◽  
Polymerase Chain

Acid stability of Bacillus licheniformis alpha amylase (BLA) was improved by error-prone polymerase chain reaction. The mutated BLA gene was expressed in Escherichia coli. An acid stability double mutant (K344R/H405R in BLA) was isolated. Two single mutants K344R and H405R were obtained by the way of site-directed mutagenesis. The enzymes (BLA) of the three mutants were isolated and characterized. Kinetic studies showed that the kcat/Km values of the mutants K344R, H405R, and K344R/H405R under pH 4.5 were about 8-, 11.5-, and 17.7-times higher than that of the wild type enzyme. As revealed by the structure models of the wild-type and mutant enzymes, the amino acids substituted of R344 and R405 in the BLA contribute to its acid stability.

Alteration of bond-cleavage pattern in the hydrolysis catalyzed by Saccharomycopsis .alpha.-amylase altered by site-directed mutagenesis

Biochemistry ◽  
1992 ◽  
Vol 31 (22) ◽  
pp. 5232-5236 ◽  
Author(s):  
Ikuo Matsui ◽  
Kazuhiko Ishikawa ◽  
Sachio Miyairi ◽  
Sakuzo Fukui ◽  
Koichi Honda
Keyword(s):  
Bond Cleavage ◽  
Site Directed Mutagenesis ◽  
Alpha Amylase ◽  
Directed Mutagenesis ◽  
Cleavage Pattern

Site-directed mutagenesis of a conserved Asn450 residue of Bacillus licheniformis γ-glutamyltranspeptidase

2016 ◽  
Vol 91 ◽  
pp. 416-425 ◽  
Author(s):  
Min-Guan Lin ◽  
Meng-Chun Chi ◽  
Yu-Yi Chen ◽  
Tzu-Fan Wang ◽  
Hui-Fen Lo ◽  
...  
Keyword(s):  
Bacillus Licheniformis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis
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