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HomeAdvanced Materials ResearchAdvanced Materials Research Vols. 774-776Enhancement of Acid Stability of Alpha Amylase...

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

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Abstract:

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 k_cat/K_m 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.

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Periodical:

Advanced Materials Research (Volumes 774-776)

Pages:

664-669

DOI:

https://doi.org/10.4028/www.scientific.net/AMR.774-776.664

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Online since:

September 2013

Authors:

Yan Jing Xu, Yi Han Liu, Shuai Fan, Fu Ping Lu

Keywords:

Acid Stability, Alpha Amylase, Bacillus licheniformis, Characterization, Structure Analysis

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© 2013 Trans Tech Publications Ltd. All Rights Reserved

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