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Molecular cloning and stress-dependent expression of a gene encoding Δ12-fatty acid desaturase in the Antarctic microalga Chlorella vulgaris NJ-7

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Abstract

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 Δ12 unsaturated fatty acids was identified which prompted us to speculate about the involvement of Δ12 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 Δ12-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 15°C and 3.9-fold at 4°C compared to the alga at 25°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 Δ12 fatty acids desaturation involved in the adaption to cold and high salinity for Antarctic C. vugaris NJ-7.

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Abbreviations

UFA:

Unsaturated fatty acid

FAD:

Fatty acid desaturases

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Acknowledgments

We thank Prof. Xu Xudong (Institute of Hydrobiology, Chinese Academy of Sciences) for providing the Antarctic microalga strain C. vulgaris NJ-7. This work was supported by the National High-Tech Research and Development Plan of China (No. 2006AA10A114), the National Key Basic Research and Development Project of China (No. 2007CB116212), the Key Innovative Project of Chinese Academy of Science (KSCX2-YW-G-002, KZCX2-YW-225), the National Natural Science Foundation of China (30670165, 40876082) and the Key Technology Research Project of Qingdao (No. 07-1-4-16-nsh).

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Authors and Affiliations

  1. Yantai Institute of Coastal Zone Research for Sustainable Development, Chinese Academy of Sciences, 264003, Yantai, People’s Republic of China

    Yandu Lu & Song Qin

  2. Institute of Oceanology, Chinese Academy of Sciences, 266071, Qingdao, People’s Republic of China

    Yandu Lu & Shaofang Liu

  3. Graduate School of the Chinese Academy of Sciences, 100049, Beijing, People’s Republic of China

    Yandu Lu & Shaofang Liu

  4. Shandong Peanut Research Institute, 266100, Qingdao, People’s Republic of China

    Xiaoyuan Chi & Qingli Yang

  5. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 266071, Qingdao, People’s Republic of China

    Zhaoxin Li

  6. Shandong Entry-Exit Inspection and Quarantine Bureau, 266001, Qingdao, People’s Republic of China

    Qinhua Gan

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  1. Yandu Lu
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  2. Xiaoyuan Chi
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  3. Qingli Yang
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  4. Zhaoxin Li
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  7. Song Qin
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Correspondence to Song Qin.

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Communicated by T. Matsunaga.

Y. Lu and X. Chi are co-first authors.

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Lu, Y., Chi, X., Yang, Q. et al. Molecular cloning and stress-dependent expression of a gene encoding Δ12-fatty acid desaturase in the Antarctic microalga Chlorella vulgaris NJ-7. Extremophiles 13, 875–884 (2009). https://doi.org/10.1007/s00792-009-0275-x

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  • DOI: https://doi.org/10.1007/s00792-009-0275-x

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