title: Intraspecific protoplast fusion ofBrettanomyces anomalusfor improved production of an extracellular β-glucosidase.
abstract: Improvement of production of an extracellular β-glucosidase with high activity byBrettanomyces anomalusPSY-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 β-glucosidase activities in liquid media were isolated. The best mutant strain showed increased cell growth in a flask culture, as well as increased β-glucosidase production. A recombinant strain, F3-25, was obtained after three rounds of genome shuffling and its production of β-glucosidase activity reached 4790 U L-1, which was a nearly eightfold increase compared to the original strainB. anomalusPSY-001. The subculture experiments indicated that F3-25 was genetically stable.
authors.affiliation: Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization, College of Life Sciences, Huanggang Normal University , Huanggang , Hubei , P. R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Wu
authors.affiliation: Key Laboratory for Green Chemical Process of Ministry of Education, School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology , Wuhan , Hubei , P. R. China.
authors.forename: Xihong
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Environment Correlative Dietology, Ministry of Education, College of Food Science and Technology, Huazhong Agricultural University , Wuhan , Hubei , P. R. China.
authors.forename: Siyi
authors.initials: S
authors.lastname: Pan
keywords: Brettanomyces anomalus PSY-001; genome shuffling; intraspecific protoplast fusion; β-glucosidase