Genetic improvement of alpha-amylase producing Bacillus licheniformis by homolog-mediated alpha-amylase gene amplification.
- Author:
Dandan NIU
1
;
Guiyang SHI
;
Zhengxiang WANG
Author Information
1. Center for Bioresource and Bioenergy, School of Biotechnology and the Key Laboratory of Industrial Biotechnology of the Ministry of Education, Jiangnan University, Wuxi 214122, China.
- Publication Type:Journal Article
- MeSH:
Bacillus;
enzymology;
genetics;
Gene Amplification;
Industrial Microbiology;
Nucleic Acid Amplification Techniques;
Transformation, Genetic;
alpha-Amylases;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2009;25(3):375-380
- CountryChina
- Language:English
-
Abstract:
Bacillus licheniformis alpha-amylase (BLA) is one of the most important enzymes involved in starch hydrolysis and many biotechnological processes. To improve the BLA productivity, an integrative plasmid pBL-amyL carrying amyL gene encoding a thermophilic alpha-amylase of B. licheniformis was constructed and transformed into B. licheniformis B0204, an industrial alpha-amylase producer. The transformants harboring different copies of amyL were developed on kanamycin by using homolog-mediated chromosomal amplification of alpha-amylase gene. The recombinants with different multiple copies of amyL integrated in the chromosome were identified by real-time PCR and evaluated by shake-flask fermentation. Recombinants harboring 2-5 multiple copies of amyL produced more alpha-amylase comparison to the parental strain B0204.
