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Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
Published on: April 1, 2016
Metabolic Engineering of Saccharomyces cerevisiae to Improve Glucan Biosynthesis
Xing Zhou1,2, Jing He1,2, Lingling Wang1,2
1The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, P.R. China.
Abstract:
β-Glucan is a chief structural polymer in the cell wall of yeast. β-Glucan has attracted intensive attention because of its wide applications in health protection and cosmetic areas. In the present study, the β-glucan biosynthesis pathway in S. Cerevisiae was engineered to enhance β-glucan accumulation. A newly identified bacterial β-1, 6-glucan synthase GsmA from Mycoplasma agalactiae was expressed, and increased β-glucan content by 43%. In addition, other pathway enzymes were investigated to direct more metabolic flux towards the building of β-glucan chains. We found that overexpression of Pgm2 (phosphoglucomutase) and Rho1 (a GTPase for activating glucan synthesis) significantly increased β-glucan accumulation. After further optimization of culture conditions, the β-glucan content was increased by 53.1%. This study provides a new approach to enhance β-glucan biosynthesis in Saccharomyces cerevisiae.
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