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Molecular cloning and stress-dependent expression of a gene encoding Delta(12)-fatty acid desaturase in the Antarctic microalga Chlorella vulgaris NJ-7

文献类型： 外文期刊

作者： Lu, Yandu 1 ; Chi, Xiaoyuan 4 ; Yang, Qingli 4 ; Li, Zhaoxin 5 ; Liu, Shaofang 2 ; Gan, Qinhua 6 ; Qin, Song 1 ;

作者机构： 1.Chinese Acad Sci, Yantai Inst Coastal Zone Res Sustainable Dev, Yantai 264003, Peoples R China

2.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China

3.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China

4.Shandong Peanut Res Inst, Qingdao 266100, Peoples R China

5.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China

6.Shandong Entry Exit Inspect & Quarantine Bur, Qingdao 266001, Peoples R China

关键词： Cold stress;High salinity stress;Antarctic;Delta(12) fatty acid desaturases

期刊名称：EXTREMOPHILES （ 影响因子：2.395； 五年影响因子：2.668 ）

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收录情况： SCI

摘要： The psychrotrophic Antarctic alga, Chlorella vulgaris NJ-7, grows under an extreme environment of low temperature and high salinity. In an effort to better understand the correlation between fatty acid metabolism and acclimation to Antarctic environment, we analyzed its fatty acid compositions. An extremely high amount of Delta(12) unsaturated fatty acids was identified which prompted us to speculate about the involvement of Delta(12) fatty acid desaturase in the process of acclimation. A full-length cDNA sequence, designated CvFAD2, was isolated from C. vulgaris NJ-7 via reverse transcription polymerase chain reaction (RT-PCR) and RACE methods. Sequence alignment and phylogenetic analysis showed that the gene was homologous to known microsomal Delta(12)-FADs with the conserved histidine motifs. Heterologous expression in yeast was used to confirm the regioselectivity and the function of CvFAD2. Linoleic acid (18:2), normally not present in wild-type yeast cells, was detected in transformants of CvFAD2. The induction of CvFAD2 at an mRNA level under cold stress and high salinity is detected by real-time PCR. The results showed that both temperature and salinity motivated the upregulation of CvFAD2 expression. The accumulation of CvFAD2 increased 2.2-fold at 15A degrees C and 3.9-fold at 4A degrees C compared to the alga at 25A degrees C. Meanwhile a 1.7- and 8.5-fold increase at 3 and 6% NaCl was detected. These data suggest that CvFAD2 is the enzyme responsible for the Delta(12) fatty acids desaturation involved in the adaption to cold and high salinity for Antarctic C. vugaris NJ-7.

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