[枯草芽孢杆菌168中肝素聚糖合成途径的优化]

[Optimization of heparosan synthetic pathway in Bacillus subtilis 168].

作者信息

Zhang Linpei, Wang Hao, Zhou Zhengxiong, Du Guocheng, Chen Jian, Kang Zhen

机构信息

Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu, China.

School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2017 Jun 25;33(6):936-945. doi: 10.13345/j.cjb.160453.

DOI:10.13345/j.cjb.160453

PMID:28895355

Abstract

Heparosan is the start point for chemoenzymatic synthesis of heparin and it is of great significance to efficiently synthesize heparosan in microorganisms. The effects of overexpressing key enzyme genes of the UDP-glucuronic acid (UDP-GlcUA) pathway (pgcA, gtaB and tuaD) or the UDP-N-acetyl-glucosamine (UDP-GlcNAc) pathway (glmS, glmM and glmU) on the heparosan production and molecular mass were analyzed in the constructed heparosan-producing Bacillus subtilis ((1.71±0.08) g/L). On this basis, heparosan production was increased to (2.89±0.11) g/L with the molecular mass of (75.90±1.18) kDa through co-overexpressing the tuaD, gtaB, glmU, glmM and glmS genes in shake flask cultivation. In the 3 L fed-batch fermentation, heparosan production was improved to (7.25±0.36) g/L with the molecular mass of (46.66±2.71) kDa, providing the potential for heparosan industrial production.

摘要

乙酰化硫酸肝素是化学酶法合成肝素的起始点，在微生物中高效合成乙酰化硫酸肝素具有重要意义。在构建的产乙酰化硫酸肝素枯草芽孢杆菌（(1.71±0.08) g/L）中，分析了过表达尿苷二磷酸葡萄糖醛酸（UDP-GlcUA）途径（pgcA、gtaB和tuaD）或尿苷二磷酸-N-乙酰葡糖胺（UDP-GlcNAc）途径（glmS、glmM和glmU）的关键酶基因对乙酰化硫酸肝素产量和分子量的影响。在此基础上，通过在摇瓶培养中共同过表达tuaD、gtaB、glmU、glmM和glmS基因，乙酰化硫酸肝素产量提高到(2.89±0.11) g/L，分子量为(75.90±1.18) kDa。在3 L补料分批发酵中，乙酰化硫酸肝素产量提高到(7.25±0.36) g/L，分子量为(46.66±2.71) kDa，为乙酰化硫酸肝素的工业化生产提供了潜力。