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

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

作　　者：时韶磊 林红[1,2,3] 舒正玉[1,2,3] 刘艳如[1,2,3] 李欣[1,2,3] 武海龙[1,2,3] 黄建忠[1,2,3] 

机构地区：[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

出　　处：《生物技术通报》2014年第3期123-129,共7页Biotechnology Bulletin

基　　金：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)

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-ΔlipA菌株的细胞结合脂肪酶活性降低了42%。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%.

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 

分 类 号：Q936[生物学—微生物学]