木糖酸氧化途径在枯草芽孢杆菌的构建及转化乙醇酸的研究

doi:10.13560/j.cnki.biotech.bull.1985.2019-0038

生物技术通报 ›› 2019, Vol. 35 ›› Issue (6): 76-82.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0038

木糖酸氧化途径在枯草芽孢杆菌的构建及转化乙醇酸的研究

凌翓^1,2, 纪明华^1,2, 段海燕^1,3, 史吉平^1,2,3, 孙俊松^1,2,3

1. 中国科学院上海高等研究院,上海 201210;
2. 中国科学院大学,北京 100049;
3. 上海科技大学,上海 201210

收稿日期:2019-01-09 出版日期:2019-06-26 发布日期:2019-07-08
作者简介:凌翓,女,硕士研究生,研究方向：微生物遗传工程;E-mail：lingxie@sari.ac.cn
基金资助:
福建省STS 合作项目（2018T3021）

Construction of An Oxidation Pathway of Xylonic Acid in Bacillus subtilis for Production of Glycolic Acid

LING Xie^1,2, JI Ming-hua^1,2, DUAN Hai-yan^1,3, SHI Ji-ping^1,2,3, SUN Jun-song^1,2,3

1. Shanghai Advanced Research Institute,Chinese Academy of Sciences,Shanghai 201210;
2. University of Chinese Academy of Sciences,Beijing 100049;
3. Shanghai Tech University,Shanghai 201210

Received:2019-01-09 Published:2019-06-26 Online:2019-07-08

摘要/Abstract

摘要： 通过过表达外源基因,旨在获得一株生产乙醇酸的食品安全菌。通过构建质粒在枯草芽孢杆菌A164S中过表达来自大肠杆菌的木糖酸降解途径,并用高效液相色谱对产物进行检测。诱导质粒表达后,重组枯草芽孢杆菌菌株成功地将木糖酸转化为乙醇酸,摩尔转化率达到42.54%。同时,枯草芽孢杆菌自身因存在非特异的醛缩酶及乙二醛脱氢酶,能够在只表达木糖酸脱水酶YjhG的情况下获得生物转化木糖酸的能力。成功构建了一株可利用木糖酸生产乙醇酸的枯草芽孢杆菌,并发现了可能的YjhH同工酶的存在。

关键词: 枯草芽孢杆菌, 乙醇酸, 木糖酸, 木糖酸脱水酶

Abstract: It is aimed that a strain of a food-safety bacterium producing glycolic acid can be obtained by over-expressing foreign genes. An over-expression system comprising of xylonate degradation pathway from Escherichia coli was constructed and introduced into Bacillus subtilis（A164S）,glycolic acid was detected as the main product in the broth of recombinant B. subtilis,using high performance liquid chromatography（HPLC）,and the conversion rate of xylonite to glycolic acid reached 42.54%. Meanwhile,Bacillus subtilis,because of its own nonspecific aldolase and glyoxal dehydrogenase,obtained the ability to biologically transform xylic acid under the condition that only xylose dehydratase YjhG was expressed. In conclusion,a strain of B. subtilis producing glycolic acid from xylic acid is successfully constructed and the possible existence of YjhH isozyme is uncovered.

Key words: Bacillus subtilis, glycolic acid, xylonic acid, xylonate dehydratase

凌翓, 纪明华, 段海燕, 史吉平, 孙俊松. 木糖酸氧化途径在枯草芽孢杆菌的构建及转化乙醇酸的研究[J]. 生物技术通报, 2019, 35(6): 76-82.

LING Xie, JI Ming-hua, DUAN Hai-yan, SHI Ji-ping, SUN Jun-song. Construction of An Oxidation Pathway of Xylonic Acid in Bacillus subtilis for Production of Glycolic Acid[J]. Biotechnology Bulletin, 2019, 35(6): 76-82.

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木糖酸氧化途径在枯草芽孢杆菌的构建及转化乙醇酸的研究

Construction of An Oxidation Pathway of Xylonic Acid in Bacillus subtilis for Production of Glycolic Acid

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