Effect of Nitrogen Sources on Omega-3 Polyunsaturated Fatty Acid Biosynthesis and Gene Expression in Thraustochytriidae sp.

被引:12
作者
Li, Siting [1 ,2 ,3 ,4 ]
Hu, Zhangli [1 ,3 ,4 ]
Yang, Xuewei [1 ,3 ,4 ]
Li, Yan [2 ]
机构
[1] Shenzhen Univ, Coll Life Sci & Oceanog, Guangdong Technol Res Ctr Marine Algal Bioengn, Guangdong Key Lab Plant Epigenet, Shenzhen 518060, Peoples R China
[2] Auckland Univ Technol, Fac Hlth & Environm Sci, Sch Sci, Auckland 1142, New Zealand
[3] Shenzhen Univ, Coll Life Sci & Oceanog, Shenzhen Key Lab Marine Biol Resources & Ecol Env, Shenzhen Key Lab Microbial Genet Engn, Shenzhen 518055, Peoples R China
[4] Shenzhen Univ, Longhua Innovat Inst Biotechnol, Shenzhen 518060, Peoples R China
关键词
nitrogen source; Thraustochytriidae sp; n-3 polyunsaturated fatty acids; elongation and unsaturation; reference genes; DOCOSAHEXAENOIC ACID; NITRATE ASSIMILATION; COA HYDRATASE; ACCUMULATION; STRESS; DHA; OMEGA-3-FATTY-ACIDS; OVEREXPRESSION; IDENTIFICATION; STARVATION;
D O I
10.3390/md18120612
中图分类号
R914 [药物化学];
学科分类号
100701 ;
摘要
The molecular mechanism that contributes to nitrogen source dependent omega-3 polyunsaturated fatty acid (n-3 PUFA) synthesis in marine oleaginous protists Thraustochytriidae sp., was explored in this study. The fatty acid (FA) synthesis was significantly influenced by the supplement of various levels of sodium nitrate (SN) (1-50 mM) or urea (1-50 mM). Compared with SN (50 mM) cultivation, cells from urea (50 mM) cultivation accumulated 1.16-fold more n-3 PUFAs (49.49% docosahexaenoic acid (DHA) (w/w, of total FAs) and 5.28% docosapentaenoic acid (DPA) (w/w, of total FAs)). Strikingly higher quantities of short chain FAs (<18 carbons) (52.22-fold of that in urea cultivation) were produced from SN cultivation. Ten candidate reference genes (RGs) were screened by using four statistical methods (geNorm, NormFinder, Bestkeeper and RefFinder). MFT (Mitochondrial folate transporter) and NUC (Nucleolin) were determined as the stable RGs to normalize the RT-qPCR (real-time quantitative polymerase chain reaction) data of essential genes related to n-3 PUFAs-synthesis. Our results elucidated that the gene transcripts of delta(3,5)-delta(2,4)-dienoyl-CoA isomerase, enoyl-CoA hydratase, fatty acid elongase 3, long-chain fatty acid acyl-CoA ligase, and acetyl-CoA carboxylase were up-regulated under urea cultivation, contributing to the extension and unsaturated bond formation. These findings indicated that regulation of the specific genes through nitrogen source could greatly stimulate n-3 PUFA production in Thraustochytriidae sp.
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页数:17
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