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Enhanced Production of Bacterial Cellulose in Komagataeibacter xylinus Via Tuning of Biosynthesis Genes with Synthetic RBS

Journal of Microbiology and Biotechnology 2020년 30권 9호 p.1430 ~ 1435
Hur Dong-Hoon, 최우성, 김태용, 이상엽, 박진환, Jeong Ki-Jun,
소속 상세정보
 ( Hur Dong-Hoon ) - Korea Advanced Institute of Science and Technology Department of Chemical and Biomolecular Engineering
최우성 ( Choi Woo-Sung ) - Korea Advanced Institute of Science and Technology Department of Chemical and Biomolecular Engineering
김태용 ( Kim Tae-Yong ) - Samsung Electronics Co. Ltd. Samsung Advanced Institute of Technology
이상엽 ( Lee Sang-Yup ) - Korea Advanced Institute of Science and Technology Department of Chemical and Biomolecular Engineering
박진환 ( Park Jin-Hwan ) - Samsung Electronics Co. Ltd. Samsung Advanced Institute of Technology
 ( Jeong Ki-Jun ) - Korea Advanced Institute of Science and Technology Department of Chemical and Biomolecular Engineering

Abstract


Bacterial cellulose (BC) has outstanding physical and chemical properties, including high crystallinity, moisture retention, and tensile strength. Currently, the major producer of BC is Komagataeibacter xylinus. However, due to limited tools of expression, this host is difficult to engineer metabolically to improve BC productivity. In this study, a regulated expression system for K. xylinus with synthetic ribosome binding site (RBS) was developed and used to engineer a BC biosynthesis pathway. A synthetic RBS library was constructed using green fluorescent protein (GFP) as a reporter, and three synthetic RBSs (R4, R15, and R6) with different strengths were successfully isolated by fluorescence-activated cell sorting (FACS). Using synthetic RBS, we optimized the expression of three homologous genes responsible for BC production, pgm, galU, and ndp, and thereby greatly increased it under both static and shaking culture conditions. The final titer of BC under static and shaking conditions was 5.28 and 3.67 g/l, respectively. Our findings demonstrate that reinforced metabolic flux towards BC through quantitative gene expression represents a practical strategy for the improvement of BC productivity.

키워드

Bacterial cellulose; Komagataeibacter xylinus; RBS; fluorescence-activated cell sorting

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