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  • Title: Genetic improvement of alpha-amylase producing Bacillus licheniformis by homolog-mediated alpha-amylase gene amplification.
    Author: Niu D, Shi G, Wang Z.
    Journal: Sheng Wu Gong Cheng Xue Bao; 2009 Mar; 25(3):375-80. PubMed ID: 19621577.
    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.
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