高产二羟基丙酮重组菌株的构建

doi:10.13560/j.cnki.biotech.bull.1985.2015.08.025

生物技术通报 ›› 2015, Vol. 31 ›› Issue (8): 174-179.doi: 10.13560/j.cnki.biotech.bull.1985.2015.08.025

高产二羟基丙酮重组菌株的构建

许晓菁^{1, 2}, 赵琼², 王祥河²

1.天津市产品质量监督检测技术研究院,天津 300380; 2.天津市工业微生物研究所,天津 300462

收稿日期:2014-12-09 出版日期:2015-08-21 发布日期:2015-08-22
作者简介:许晓菁,女,博士,高级工程师,研究方向：工业微生物发酵及检测方法开发;E-mail：xxjmiss@163.com
基金资助:
天津市应用基础与前沿技术研究计划（天津市自然科学基金）重点项目（10JCZDJC16600）

Construction of a Recombinant Strain Producing High-yield Dihydroxyacetone

Xu Xiaojing^{1, 2}, Zhao Qiong¹, Wang Xianghe¹

1. Industrial Microbe Research Institute of Tianjin,Tianjin 300462; 2. Tianjin Product Quality Inspection Technology Research Institute,Tianjin 300380

Received:2014-12-09 Published:2015-08-21 Online:2015-08-22

摘要/Abstract

摘要： 旨在构建一株过量表达编码膜系甘油脱氢酶的sldAB基因的重组菌株,以提高二羟基丙酮产量。以氧化葡萄糖酸杆菌ATCC621H的基因组DNA为模板,运用PCR方法扩增得到基因sldAB,并连接到pBBR1MCS-2质粒上,构建表达载体pBBR1MCS-2-sldAB。通过电转化将载体pBBR1MCS-2-sldAB转化进入氧化葡萄糖酸杆菌ATCC621H内,得到重组菌株GOX205。结果显示,重组菌株构建成功,其甘油脱氢酶的酶活力较之于出发菌株提高了26%。在甘油初始浓度100 g/L的甘油发酵培养基中,较之于出发菌株,GOX205的生长状况良好,发酵52 h时DHA浓度达到94.1 g/L,较之于出发菌株提高了19.7%,甘油残量降低了15.1 g/L。

关键词: 1, 3-二羟基丙酮, 重组菌株, 甘油, 过量表达

Abstract: This work aims to construct a recombinant strain containing gene sldAB over-expressing membrane-bound glycerol dehydrogenase for raising the yield of dihydroxyacetone（DHA）. Firstly, using genomic DNA of Gluconobacter oxydans ATCC621H as a template, sldAB amplified by PCR was ligated to the plasmid pBBR1MCS-2 and the expression vector pBBR1MCS-2-sldAB was constructed;subsequently the plasmid pBBR1MCS-2-sldAB was transformed into the G. oxydans ATCC621H by electrotransfer, and the recombinant strain GOX205 was obtained. Results showed that the recombinant strain was constructed successfully, and the activity of glycerol dehydrogenase increased by 26% compared with the original strain. When initial concentration 100 g/L of glycerol as carbon source, the growth of GOX205 was significantly improved compared with the original strain, the final DHA concentration was 94.1 g/L and increased by 19.7%, and the residue of glycerol decreased 15.1 g/L. Therefore, this study provides a basis for further improving the biotransformation process of DHA production.

Key words: 1, 3-dihydroxyacetone, recombinant strain, glycerol, over-expression

许晓菁, 赵琼, 王祥河. 高产二羟基丙酮重组菌株的构建[J]. 生物技术通报, 2015, 31(8): 174-179.

Xu Xiaojing, Zhao Qiong, Wang Xianghe. Construction of a Recombinant Strain Producing High-yield Dihydroxyacetone[J]. Biotechnology Bulletin, 2015, 31(8): 174-179.

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高产二羟基丙酮重组菌株的构建

Construction of a Recombinant Strain Producing High-yield Dihydroxyacetone

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