通过增加编码脂肪酸去饱和酶和延长酶的基因拷贝数来提高毕赤酵母中花生四烯酸和二十碳五烯酸的产量。

Improvement of arachidonic acid and eicosapentaenoic acid production by increasing the copy number of the genes encoding fatty acid desaturase and elongase into Pichia pastoris.

作者信息

Li Yun-Tao, Li Mao-Teng, Fu Chu-Hua, Zhou Peng-Peng, Liu Jian-Min, Yu Long-Jiang

机构信息

Institute of Resource Biology and Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China.

出版信息

Biotechnol Lett. 2009 Jul;31(7):1011-7. doi: 10.1007/s10529-009-9970-z. Epub 2009 Mar 22.

DOI:10.1007/s10529-009-9970-z

PMID:19306085

Abstract

Genes encoding Delta6 desaturase, Delta6 fatty acid elongase, and Delta5 desaturase from the alga, Phaeodactylum tricornutum, were co-expressed in Pichia pastoris to produce arachidonic acid (ARA; 20:4 Delta(5, 8, 11, 14)) and eicosapentaenoic acid (EPA; 20:5 Delta(5, 8, 11, 14, 17)). A panel of Pichia clones carrying progressively increasing copies of the heterologous gene expression cassette was created using an in vitro multimerization approach. ARA and EPA accumulated up to 0.3 and 0.1% of total fatty acids, respectively, in the recombinant P. pastoris carrying with double copies of these three heterologous genes, as compared to 0.1 and 0.05%, respectively, in the recombinant P. pastoris with single copy.

摘要

将来自三角褐指藻的编码Δ6去饱和酶、Δ6脂肪酸延长酶和Δ5去饱和酶的基因在毕赤酵母中进行共表达，以生产花生四烯酸（ARA；20:4Δ(5, 8, 11, 14)）和二十碳五烯酸（EPA；20:5Δ(5, 8, 11, 14, 17)）。使用体外多聚化方法构建了一组携带逐渐增加的异源基因表达盒拷贝数的毕赤酵母克隆。在携带这三个异源基因双拷贝的重组毕赤酵母中，ARA和EPA分别累积至总脂肪酸的0.3%和0.1%，相比之下，在携带单拷贝的重组毕赤酵母中分别为0.1%和0.05%。