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Identification and characterization of a novel bifunctional Δ12/Δ15-fatty acid desaturase gene from Rhodosporidium kratochvilovae

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Abstract

Objectives

To elucidate the biosynthesis pathway of linoleic acid and α-linolenic acid in Rhodosporidium kratochvilovae YM25235 and investigate the correlation of polyunsaturated fatty acids with its cold adaptation.

Results

A 1341 bp cDNA sequence, designated as RKD12, putatively encoding a Δ12-desaturase was isolated from YM25235. Sequence analysis indicated that this sequence comprised a complete ORF encoding 446 amino acids of 50.6 kDa. The encoded amino acid sequence shared higher similarity to known fungal Δ12-desaturases that are characteristic of three conserved histidine-rich motifs. RKD12 was further transformed into Saccharomyces cerevisiae INVScl for functional characterization. Fatty acid analysis showed the yeast transformants accumulated two new fatty acids: linoleic acid and α-linolenic acid. Furthermore, mRNA expression level of RKD12 and the content of linoleic acid and α-linolenic acid were increased significantly with the culture temperature downshift from 30 to 15 °C, which might be helpful for the cold adaptation of YM25235.

Conclusion

RKD12 is a novel bifunctional ∆12/∆15-desaturase gene, and the increased RKD12 mRNA expression level and PUFAs content at low temperature might be helpful for the cold adaptation of YM25235.

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Acknowledgments

We acknowledge Professor Shaolan Li in Yunnan Institute of Microbiology, Yunnan University, for her kind present of Rhodosporidium kratochvilovae strain YM25235. This study was supported by the National Natural Science Foundation of China (Grant Nos. 31160016 and 31260034).

Supporting Information

Supplementary Figure 1—Sequence alignment of deduced amino acids of the R. kratochvilovae Δ12-desaturase (RAD12) with the Δ12-desaturases from Mortierella alpina IS-4 (MAD12), Amylomyces rouxii (ARD12) and Komagataella pastoris (KPD12).

Supplementary Figure 2—GC analysis of fatty acid compositions from YM25235 grown at 28 °C and 15 °C.

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Authors and Affiliations

  1. Faculty of Life Science and Technology, Kunming University of Science and Technology, 727 South Jingming Road, Kunming, 650500, China

    Jinjin Cui, Shiwu He, Xiuling Ji, Lianbing Lin, Yunlin Wei & Qi Zhang

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  1. Jinjin Cui
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  2. Shiwu He
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  3. Xiuling Ji
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  4. Lianbing Lin
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  5. Yunlin Wei
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  6. Qi Zhang
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Correspondence to Qi Zhang.

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Cui, J., He, S., Ji, X. et al. Identification and characterization of a novel bifunctional Δ12/Δ15-fatty acid desaturase gene from Rhodosporidium kratochvilovae . Biotechnol Lett 38, 1155–1164 (2016). https://doi.org/10.1007/s10529-016-2090-7

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