Journal
BIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT
Volume 28, Issue 5, Pages 878-881Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/13102818.2014.955290
Keywords
beta-glucosidase; genome shuffling; intraspecific protoplast fusion; Brettanomyces anomalus PSY-001
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Funding
- Huazhong Agricultural University
- Huanggang Normal University
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Improvement of production of an extracellular beta-glucosidase with high activity by Brettanomyces anomalus PSY-001 was performed by using recursive protoplast fusion in a genome-shuffling format. The initial population was generated by ultraviolet irradiation, ultrasonic mutagenesis and, then, subjected to recursive protoplast fusion. Mutant strains exhibiting significantly higher beta-glucosidase activities in liquid media were isolated. The best mutant strain showed increased cell growth in a flask culture, as well as increased beta-glucosidase production. A recombinant strain, F3-25, was obtained after three rounds of genome shuffling and its production of beta-glucosidase activity reached 4790U L-1, which was a nearly eightfold increase compared to the original strain B. anomalus PSY-001. The subculture experiments indicated that F3-25 was genetically stable.
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