Burkholderia sp. ZYB002中lipA基因的敲除对细胞脂肪酶活性的影响

生物技术通报 ›› 2014, Vol. 0 ›› Issue (3): 123-129.

Burkholderia sp. ZYB002中lipA基因的敲除对细胞脂肪酶活性的影响

时韶磊林红舒正玉刘艳如李欣武海龙黄建忠

(1.福建师范大学工业微生物发酵技术国家地方联合工程研究中心，福州 350108；2.福建师范大学教育部工业微生物工程中心，福州 350108；3. 福建师范大学生命科学学院，福州 350108)

收稿日期:2013-12-16 出版日期:2014-03-29 发布日期:2014-03-31
作者简介:时韶磊，男，研究方向:酶工程，E-mail:429490879@qq.com；林红同为本文第一作者
基金资助:
国家自然科学基金项目(31370802)，福建省科技厅重点项目(2013H0021)，福建省自然科学基金杰青项目(2009J06013)

Effect of the lipA Inactivation on the Cell-bound Lipase Yield of Burkholderia sp. ZYB002

Shi Shaolei, Lin Hong, Shu Zhengyu, Liu Yanru, Li Xin, Wu Hailong, Huang Jianzhong

(1. National & Local United Engineering Research Center of Industrial Microbiology and Fermentation Technology, Ministry of Education, Fujian Normal University, Fuzhou 350108；2. Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, Fuzhou 350108；3. College of Life Sciences, Fujian Normal University, Fuzhou 350108)

Received:2013-12-16 Published:2014-03-29 Online:2014-03-31

摘要/Abstract

摘要： Burkholderia sp. ZYB002菌株不仅能产生大量的胞外脂肪酶，还能产生大量的细胞结合脂肪酶。对细胞结合脂肪酶的种类进行定性研究，将为开发全细胞脂肪酶催化剂奠定基础。通过分析与 Burkholderia sp. ZYB002菌株具有较高同源性的 Burkholderia cepacia J2315菌株的脂肪酶基因家族，推测潜在的细胞结合脂肪酶编码基因。通过同源重组插入失活 Burkholderia sp. ZYB002菌株的 lipA基因，筛选出突变体转化子；测定突变体菌株细胞结合脂肪酶的活性，并与野生型菌株比较。试验结果表明， lipA基因插入失活的 Burkholderia sp. ZYB002-Δ lipA菌株的细胞结合脂肪酶活性降低了42%。

关键词: 伯克霍尔德菌ZYB002, 脂肪酶LipA, 细胞结合脂肪酶, 基因敲除, 三亲本杂交

Abstract: Burkholderia sp. ZYB002 strain can produce large amount of extracellular lipase and cell-bound lipase. The type of the cell-bound lipase from Burkholderia sp. ZYB002 was qualitatively analyzed, which will help to develop the whole cell lipase in the organic synthesis. Series of potential cell-bound lipase genes were presumed from the known genomic DNA information of Burkholderia cepacia J2315, which was highly homologous to that of Burkholderia sp. ZYB002. The lipA gene from Burkholderia sp. ZYB002 was selected and then inactivated by homologous recombination. The mutant with lipA inactivation was screened and the cell-bound lipase activity from Burkholderia sp. ZYB002-Δ lipA was tested. The cell-bound lipase activity from Burkholderia sp. ZYB002-Δ lipA decreased by 42%.

Key words: Burkholderia, sp. ZYB002 Extracellular lipase A, Cell-bound lipase, Gene knockout, Triparental mating

时韶磊林红舒正玉刘艳如李欣武海龙黄建忠. Burkholderia sp. ZYB002中lipA基因的敲除对细胞脂肪酶活性的影响[J]. 生物技术通报, 2014, 0(3): 123-129.

Shi Shaolei, Lin Hong, Shu Zhengyu, Liu Yanru, Li Xin, Wu Hailong, Huang Jianzhong^{. Effect of the lipA Inactivation on the Cell-bound Lipase Yield of Burkholderia sp. ZYB002[J]. Biotechnology Bulletin, 2014, 0(3): 123-129.}

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