Production of conjugated linoleic acid by heterologous expression of linoleic acid isomerase in oleaginous fungus Mortierella alpina
Production of conjugated linoleic acid by heterologous expression of linoleic acid isomerase in oleaginous fungus Mortierella alpina
复制标题
产油真菌高山被孢霉异源表达亚油酸异构酶生产共轭亚油酸
DOI：
10.1007/s10529-015-1871-8
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发表时间：
2015-10-01
影响因子：
2.7
通讯作者：
Chen, Yong Q.
中科院分区：
文献类型：
--
作者：
Hao, Danhui;Chen, Haiqin;Chen, Yong Q.
目的：通过将反式10，顺式12共轭亚油酸（CLA）引入到产油真菌高山被孢霉的多不饱和脂肪酸（PUFA）中，提高其商品价值。结果：成功地将派基因和密码子优化后的派基因在M. alpina中异源表达。通过根癌农杆菌介导的转化（ATMT）转化高山高山云杉。编码使用的修改显着提高派转录的翻译和反式-10，顺式-12共轭亚油酸产生高达1.2毫克l（-1），这相当于约。0.05%的总脂肪酸（TFA）。由于派更喜欢游离亚油酸作为底物，而不是任何其他形式，因此添加5 μM长链酰基CoA合成酶抑制剂，反式-10，顺式-12 CLA含量增加约10%。24-倍至29 mg l（-1），达到M中TFA的1.2%（w/w）。结论：派在M. alpina中呈异质性表达。利用ATMT法合成高浓度CLA是一条可行的途径，该系统为CLA的工业化生产提供了一个可行的平台。
Objective:To increase the commercial value of oleaginous fungus Mortierella alpina by incorporation of trans-10,cis-12 conjugated linoleic acid (CLA) into the polyunsaturated fatty acids (PUFAs) of M. alpina via Propionibacterium acnes isomerase (PAI) conversion.Results:The PAI gene and the codon optimization version were heterologously expressed in M. alpina via Agrobacterium tumefaciens-mediated transformation (ATMT). Coding usage modification significantly improved the translation of PAI transcripts and trans-10,cis-12 CLA was produced up to 1.2 mg l(-1), which corresponds to approx. 0.05% of the total fatty acid (TFA). Since PAI prefers free linoleic acid as a substrate rather than any other forms, 5 μM long-chain acyl CoA synthetase inhibitor was added and the trans-10,cis-12 CLA content increased approx. 24-fold to 29 mg l(-1), reaching up to 1.2% (w/w) of the TFA in M. alpina.Conclusion:Heterologous expression of PAI in M. alpina by ATMT methods is a practicable way in biosynthesis of CLA and this system may be a feasible platform for industrial production of CLA.