基因敲除弱化产1,3-丙二醇Klebsiella pneumoniae荚膜多糖的合成

doi:10.16085/j.issn.1000-6613.2017-0081

化工进展 ›› 2017, Vol. 36 ›› Issue (09): 3447-3452.DOI: 10.16085/j.issn.1000-6613.2017-0081

基因敲除弱化产1,3-丙二醇Klebsiella pneumoniae荚膜多糖的合成

刘情^1,2, 王小婉², 诸葛斌^1,2, 陆信曜^1,2, 宗红^1,2, 方慧英^1,2, 宋健³

1 江南大学糖化学与生物技术教育部重点实验室, 江苏无锡 214122;
2 江南大学生物工程学院, 工业生物技术教育部重点实验室工业微生物研究中心, 江苏无锡 214122;
3 江南大学化学与材料工程学院, 江苏无锡 214122

收稿日期:2017-01-13 修回日期:2017-02-08 出版日期:2017-09-05 发布日期:2017-09-05
通讯作者: 诸葛斌,博士,教授。
作者简介:刘情(1991-),女,硕士研究生,研究方向为发酵工程。E-mail:qingliu19@163.com
基金资助:
高等学校学科创新引智计划（111-2-06）和江苏省青年自然科学基金（BK20140134）项目。

Reducing the capsular polysaccharide synthesis of Klebsiella pneumoniae in 1,3-propanediol fermentation by genes knocking-out

LIU Qing^1,2, WANG Xiaowan², ZHUGE Bin^1,2, LU Xinyao^1,2, ZONG Hong^1,2, FANG Huiying^1,2, SONG Jian³

1 Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu, China;
2 The KeyLaboratory of Industrial Biotechnology, Ministry of Education, Research Centre of Industrial Microbiology, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China;
3 School of Chemistry and Material, Jiangnan University, Wuxi 214122, Jiangsu, China

Received:2017-01-13 Revised:2017-02-08 Online:2017-09-05 Published:2017-09-05

摘要/Abstract

摘要： 1，3-丙二醇（1，3-PDO）是重要的平台化合物，在高性能纤维合成等领域具有广泛应用。Klebsiella pneumoniae是优良的1，3-PDO生物合成细胞工厂，但该菌代谢甘油合成1，3-PDO时积累大量荚膜多糖，影响底物与产物的高效转运，导致发酵液黏度增大，限制发酵法生产1，3-丙二醇的下游提取及产业化。本研究基于已报道的K.pneumoniae荚膜合成途径，利用Red重组技术对K.pneumoniae荚膜多糖合成途径中的起始糖基转移酶wecA基因及参与荚膜多糖后期组装的跨膜蛋白wzi基因进行敲除，并考察上述基因缺失对菌株荚膜含量、细胞形态、菌体生长、发酵液黏度及产物合成的影响。研究发现，同时缺失wecA和wzi基因对于弱化荚膜多糖效果显著，荚膜含量降低38.5%，细胞基本丧失菌毛合成及相互粘连能力，1，3-PDO产量提高23.0%，为菌株改造提供了一种新思路。

关键词: 荚膜多糖, 敲除, Klebsiella pneumoniae, 1, 3-丙二醇

Abstract: 1,3-propanediol (1,3-PDO)is an important platform compound and has been widely used in the production of high performance fiber synthesis. Klebsiella pneumoniae is an excellent 1,3-PDO biosynthetic cell factory. However,the capsular polysaccharide accumulation of K. pneumoniae increased the viscosity of fermentation broth and impacted the efficient transport of substrate and product,resulting in an obstacle in downstream extraction and industrialization of 1,3-PDO. In this study,by analyzing the capsular synthesis pathway of K. pneumoniae,the key genes wecA (initial glycosyltransferase gene) and wzi involved in the assembly of capsular polysaccharide were disrupted by Red recombinant system. The effects of genes deletion on capsule content,cell morphology,cell growth,broth fluid viscosity,and product synthesis,were studied. The capsular content was reduced by 38.5%,which led to the loss of cells fimbriae synthesis ability and the adhesion,and the titer of 1,3-PDO increased by 23.0%. The study provided a novel prospect on the development of K. pneumoniae on the production of 1,3-PDO.

Key words: capsular polysaccharide, knocking-out, Klebsiella pneumoniae, 1,3-propanediol

中图分类号:

刘情, 王小婉, 诸葛斌, 陆信曜, 宗红, 方慧英, 宋健. 基因敲除弱化产1,3-丙二醇Klebsiella pneumoniae荚膜多糖的合成[J]. 化工进展, 2017, 36(09): 3447-3452.

LIU Qing, WANG Xiaowan, ZHUGE Bin, LU Xinyao, ZONG Hong, FANG Huiying, SONG Jian. Reducing the capsular polysaccharide synthesis of Klebsiella pneumoniae in 1,3-propanediol fermentation by genes knocking-out[J]. Chemical Industry and Engineering Progress, 2017, 36(09): 3447-3452.

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基因敲除弱化产1,3-丙二醇Klebsiella pneumoniae荚膜多糖的合成

Reducing the capsular polysaccharide synthesis of Klebsiella pneumoniae in 1,3-propanediol fermentation by genes knocking-out

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