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Engineering a newly isolated Bacillus licheniformis strain for the production of (2R,3R)-butanediol.
Journal of Industrial Microbiology & Biotechnology ( IF 3.2 ) Pub Date : 2019-11-22 , DOI: 10.1007/s10295-019-02249-4
Chan Woo Song 1 , Rathnasingh Chelladurai 1 , Jong Myoung Park 1 , Hyohak Song 1
Affiliation  

Several microorganisms can produce 2,3-butanediol (BDO), an industrially promising chemical. In this study, a Bacillus licheniformis named as 4071, was isolated from soil sample. It is a GRAS (generally recognized as safe) strain and could over-produce 2,3-BDO. Due to its mucoid forming characteristics, UV-random mutagenesis was carried out to obtain a mucoid-free strain, 4071-15. As a result, capabilities of 4071-15 strain in terms of transformation efficiency of bacillus plasmids (pC194, pUB110, and pUCB129) and fermentation performance were highly upgraded compared to those of the parent strain. In particular, 4071-15 strain could produce 123 g/L of 2,3-BDO in a fed-batch fermentation in which the ratio of (2R,3S)- to (2R,3R)-form isomers was 1:1. To increase the selectivity of (2R,3R)-BDO, budC gene was deleted by using temperature-sensitive gene deletion process via homologous recombination. The 4071-15 △budC mutant strain dramatically increased selectivity of (2R,3R)-BDO to 91% [96.3 g/L of (2R,3R)-BDO and 9.33 g/L of (2R,3S)-BDO], which was 43% higher than that obtained by the parent strain. This study has shown the potential of an isolate for 2,3-BDO production, and that the ratio of 2,3-BDO can be controlled by genetic engineering depending on its industrial usage.

中文翻译:

工程新分离的地衣芽孢杆菌菌株,用于生产(2R,3R)-丁二醇。

几种微生物可以产生工业上有希望的化学物质2,3-丁二醇(BDO)。在这项研究中,从土壤样品中分离出了一种名为4071的地衣芽孢杆菌。这是一种GRAS(通常被认为是安全的)菌株,可能会过量生产2,3-BDO。由于其粘液样的形成特性,进行了紫外线无规诱变,获得了无粘液样的菌株4071-15。结果,与亲本菌株相比,4071-15菌株在芽孢杆菌质粒(pC194,pUB110和pUCB129)的转化效率和发酵性能方面的能力大大提高。特别地,在(2R,3S)-与(2R,3R)-形式异构体之比为1:1的补料分批发酵中,4071-15菌株可产生123g / L的2,3-BDO。为了提高(2R,3R)-BDO的选择性,通过同源重组,使用温度敏感基因缺失方法缺失了budC基因。4071-15△budC突变菌株将(2R,3R)-BDO的选择性大大提高至91%[(2R,3R)-BDO为96.3 g / L和(2R,3S)-BDO为9.33 g / L],比亲本菌株获得的高43%。这项研究表明了分离出的2,3-BDO的潜力,并且2,3-BDO的比例可以通过基因工程控制,具体取决于其工业用途。
更新日期:2019-11-01
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