脂肪酸碳链延长酶与脱饱和酶在酿酒酵母中共表达  

Co-expression of Elongase and Desaturase in Saccharomyces cerevisiae

作　　者：陈晓峰[1] 江贤章[1] 毛若雨[1] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108

出　　处：《福建师范大学学报（自然科学版）》2010年第4期105-109,共5页Journal of Fujian Normal University：Natural Science Edition

基　　金：国家自然科学基金资助项目(3097004730370028);福建省自然科学基金重大项目(2003F005);福建省自然科学基金资助项目(2008F3036);福建省发展和改革委员会科技产业化项目([2005]847);福建省科技平台项目(2005Q007)

摘　　要：为了利用转基因技术在酿酒酵母Saccharomyces cerevisiae中生产二十二碳六烯酸(DHA,22∶6Δ4,7,10,13,16,19n-3),构建了同时含C20-Δ5脂肪酸碳链延长酶(TFD5)和C22-Δ4脂肪酸碳链脱饱和酶(FAD4)基因的共表达质粒pYTFD5-FAD4.该质粒是通过基因重组的方法,使脂肪酸碳链延长酶与脱饱和酶基因置于各自启动子和终止子下获得的.共表达质粒pYTFD5-FAD4转化酿酒酵母所得到基因工程菌,在添加终质量分数为2%的半乳糖,终体积分数为1%的NP-40和0.3 mmol/L的底物二十碳五烯酸(EPA,20∶5Δ5,8,11,14,17n-3)下进行诱导,可直接转化二十碳五烯酸(EPA,20∶5Δ5,8,11,14,17n-3)生成二十二碳五烯酸(DPA,22∶5Δ4,7,10,13,16n-3)和二十二碳六烯酸(DHA,22∶6Δ4,7,10,13,16,19n-3).In order to produce DHA by transgenetic technique, the recombinant plasmid pYTFDS-FAD4, including genes of delta-5 Elongase （elo5） and delta-4-Desaturase （fad4）, was constructed and transformed into Saccharomyces cerevisiae INVScl to construct recombinant strain. The genes were induced by the addition of galactose to 2%. The yeast medium was supplemented with 0.3 mmol/L exogenous fatty acid subtrate Eicosapentaenoic acid （EPA, 20 ： 5△5, 8, 11, 14, 17n-3） in the presence of 1~ NP-40. The total fatty acids were extracted from the induced cells and subjected to methyl-esterification. The fatty acid methyl esters （FAME） were analyzed by GC-MS detection. Two novel peaks corresponding to Docosapentaenoicacid （DPA, 22：5△4, 7, 10, 13, 16n-3）andDocosahexaenoicacid （DHA, 22 ： 6△4, 7, 10, 13, 16, 19n-3） methyl ester standards were detected with the same retention time which were absent in the control. The results revealed that delta-5-Elongase and delta- 4-Desaturase could specifically catalyze EPA into DPA and DHA .

关 键 词：二十二碳六烯酸 酿酒酵母 共表达 

分 类 号：Q936[生物学—微生物学]