Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Enhanced Production of Bacterial Cellulose in Komagataeibacter xylinus Via Tuning of Biosynthesis Genes with Synthetic RBS

  • Hur, Dong Hoon (Department of Chemical and Biomolecular Engineering, BK21 Plus Program, Korea Advanced Institute of Science and Technology (KAIST)) ;
  • Choi, Woo Sung (Department of Chemical and Biomolecular Engineering, BK21 Plus Program, Korea Advanced Institute of Science and Technology (KAIST)) ;
  • Kim, Tae Yong (Biomaterials Lab, Samsung Advanced Institute of Technology (SAIT), Samsung Electronics Co., Ltd.) ;
  • Lee, Sang Yup (Department of Chemical and Biomolecular Engineering, BK21 Plus Program, Korea Advanced Institute of Science and Technology (KAIST)) ;
  • Park, Jin Hwan (Biomaterials Lab, Samsung Advanced Institute of Technology (SAIT), Samsung Electronics Co., Ltd.) ;
  • Jeong, Ki Jun (Department of Chemical and Biomolecular Engineering, BK21 Plus Program, Korea Advanced Institute of Science and Technology (KAIST))
  • Received : 2020.06.18
  • Accepted : 2020.07.02
  • Published : 2020.09.28
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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.

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