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Tailoring bacterial cellulose structure through CRISPR interference-mediated downregulation of galU in Komagataeibacter xylinus CGMCC 2955
Authors:Long-Hui Huang  Qi-Jing Liu  Xue-Wen Sun  Xue-Jing Li  Miao Liu  Shi-Ru Jia  Yan-Yan Xie  Cheng Zhong
Affiliation:1. State Key Laboratory of Food Nutrition & Safety, Tianjin University of Science & Technology, Tianjin, China

Key Laboratory of Industrial Fermentation Microbiology (Ministry of Education), Tianjin University of Science & Technology, Tianjin, China;2. State Key Laboratory of Food Nutrition & Safety, Tianjin University of Science & Technology, Tianjin, China

Abstract:Diverse applications of bacterial cellulose (BC) have different requirements in terms of its structural characteristics. culturing Komagataeibacter xylinus CGMCC 2955, BC structure changes with alterations in oxygen tension. Here, the K. xylinus CGMCC 2955 transcriptome was analyzed under different oxygen tensions. Transcriptome and genome analysis indicated that BC structure is related to the rate of BC synthesis and cell growth, and galU is an essential gene that controls the carbon metabolic flux between the BC synthesis pathway and the pentose phosphate (PP) pathway. The CRISPR interference (CRISPRi) system was utilized in K. xylinus CGMCC 2955 to control the expression levels of galU. By overexpressing galU and interfering with different sites of galU sequences using CRISPRi, we obtained strains with varying expression levels of galU (3.20–3014.84%). By testing the characteristics of BC, we found that the porosity of BC (range: 62.99–90.66%) was negative with galU expression levels. However, the crystallinity of BC (range: 56.25–85.99%) was positive with galU expression levels; galU expression levels in engineered strains were lower than those in the control strains. Herein, we propose a new method for regulating the structure of BC to provide a theoretical basis for its application in different fields.
Keywords:bacterial cellulose  CGMCC 2955  CRISPR interference  CRISPR-dCas9  Komagataeibacter xylinus  transcriptome
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