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Inhibition of Polyunsaturated Fatty Acids Synthesis Decreases Growth Rate and Membrane Fluidity of Rhodosporidium kratochvilovae at Low Temperature

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Lipids

Abstract

The intention of this study was to investigate the role of polyunsaturated fatty acids (PUFA) in the cold adaptation of Rhodosporidium kratochvilovae YM25235 by knockout of the Δ12/Δ15-fatty acid desaturase gene (RKD12) to inactivate Δ12/Δ15-fatty acid desaturase. Polymerase chain reaction (PCR) amplification was used to detect the genomic structure of RKD12 gene in YM25235. The RKD12 gene was knocked out by DNA homologous recombination to inhibit the biosynthesis of PUFA. Then, the contents of linoleic acid (LNA) and α-linolenic acid (ALA) after gene knockout were investigated using a gas chromatography-mass spectrometer, followed by determination of the growth rate and membrane fluidity of YM25235 at low temperature. After PCR amplification, a 1611 bp genomic fragment was amplified from YM25235. When the RKD12 gene was knocked out, the contents of LNA and ALA in YM25235 significantly decreased. The growth rate and membrane fluidity of YM25235 decreased significantly at low temperature. Inhibition of PUFA biosynthesis by RKD12 gene knockout influenced cold adaptation of YM25235 by decreasing the PUFA content in cell membranes and reducing the growth rate and membrane fluidity of YM25235 at low temperature.

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Abbreviations

PUFA:

Polyunsaturated fatty acids

PCR:

Polymerase chain reaction

LNA:

Linoleic acid

ALA:

α-Linolenic acid

FAME:

Fatty acid methyl esters

YPD:

Yeast extract peptone dextrose

ANS:

1-Anilinonaphthalene-8-sulphonate

SD:

Standard deviation

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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

    Jun Wang, Hongjuan Nian, Xiuling Ji, Lianbing Lin, Yunlin Wei & Qi Zhang

  2. Medical faculty, Kunming University of Science and Technology, Kunming, 650500, China

    Wei Chen

Authors
  1. Jun Wang
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  2. Wei Chen
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  3. Hongjuan Nian
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  4. Xiuling Ji
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  5. Lianbing Lin
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  6. Yunlin Wei
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  7. Qi Zhang
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Corresponding author

Correspondence to Qi Zhang.

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Funding

This study was supported by a grant from the National Natural Science Foundation of China (Nos. 31160016, 31660454 and 31260034).

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The authors declare that there is no conflict of interest.

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Wang, J., Chen, W., Nian, H. et al. Inhibition of Polyunsaturated Fatty Acids Synthesis Decreases Growth Rate and Membrane Fluidity of Rhodosporidium kratochvilovae at Low Temperature. Lipids 52, 729–735 (2017). https://doi.org/10.1007/s11745-017-4273-y

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  • DOI: https://doi.org/10.1007/s11745-017-4273-y

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