State Key Laboratory of Food Nutrition & Safety, Tianjin University of Science & Technology, Tianjin 300072, PR China.
Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin 300072, PR China.
ACS Synth Biol. 2020 Nov 20;9(11):3171-3180. doi: 10.1021/acssynbio.0c00450. Epub 2020 Oct 13.
has received increasing attention as an important microorganism for the conversion of several carbon sources to bacterial cellulose (BC). However, BC productivity has been impeded by the lack of efficient genetic engineering techniques. In this study, a lambda Red and FLP/FRT-mediated site-specific recombination system was successfully established in . Using this system, the membrane bound gene , a gene that encodes glucose dehydrogenase, was knocked out to reduce the modification of glucose to gluconic acid. The engineered strain could not produce any gluconic acid and presented a decreased bacterial cellulose (BC) production due to its restricted glucose utilization. To address this problem, the gene of glucose facilitator protein (; ZMO0366) was introduced into the knockout strain coupled with the overexpression of the endogenous glucokinase gene (). The BC yield of the resultant strain increased by 63.63-173.68%, thus reducing the production cost.
已经作为一种将多种碳源转化为细菌纤维素 (BC) 的重要微生物而受到越来越多的关注。然而,由于缺乏有效的遗传工程技术,BC 的产量受到了阻碍。在本研究中,成功在 中建立了一个基于 lambda Red 和 FLP/FRT 的位点特异性重组系统。利用该系统,敲除了编码葡萄糖脱氢酶的膜结合基因 ,以减少葡萄糖向葡萄糖酸的修饰。由于葡萄糖利用受限,该工程菌株无法产生任何葡萄糖酸,导致细菌纤维素 (BC) 的产量降低。为了解决这个问题,将葡萄糖促进蛋白基因 (; ZMO0366) 引入敲除菌株中,并与内源性葡激酶基因 () 的过表达相偶联。结果菌株的 BC 产量提高了 63.63-173.68%,从而降低了生产成本。