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A novel malic enzyme gene, Mime2, from Mortierella isabellina M6-22 contributes to lipid accumulation

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Abstract

Objective

This study was aimed at cloning and characterizing a novel malic enzyme (ME) gene of Mortierella isabellina M6-22 and identifying its relation with lipid accumulation.

Methods

Mime2 was cloned from strain M6-22. Plasmid pET32aMIME2 was constructed to express ME of MIME2 in Escherichia coli BL21. After purification, the optimal pH and temperature of MIME2, as well as Km and Vmax for NADP+ were determined. The effects of EDTA or metal ions (Mn2+, Mg2+, Co2+, Cu2+, Ca2+, or Zn2+) on the enzymatic activity of MIME2 were evaluated. Besides, plasmid pRHMIME2 was created to express MIME2 in Rhodosporidium kratochvilovae YM25235, and its cell lipid content was measured by the acid-heating method. The optimal pH and temperature of MIME2 are 5.8 and 30 °C, respectively.

Results

The act ivity of MIME2 was significantly increased by Mg2+, Ca2+, or Mn2+ at 0.5 mM but inhibited by Cu2+ or Zn2+ (p < 0.05). The optimal enzymatic activity of MIME2 is 177.46 U/mg, and the Km and Vmax for NADP+ are 0.703 mM and 156.25 μg/min, respectively. Besides, Mime2 transformation significantly increased the cell lipid content in strain YM25235 (3.15 ± 0.24 vs. 2.17 ± 0.31 g/L, p < 0.01).

Conclusions

The novel ME gene Mime2 isolated from strain M6-22 contributes to lipid accumulation in strain YM25235.

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Acknowledgements

This study was funded by National Natural Science Foundation of China (Project no. 31660454 and no. 31160016).

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Author notes

  1. Shan Li and Lingyan Li should be regard as co-first authors.

Authors and Affiliations

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

    Shan Li, Lingyan Li, Xiangfeng Xiong, Xiuling Ji, Yunlin Wei, Lianbing Lin & Qi Zhang

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  1. Shan Li
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  2. Lingyan Li
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  3. Xiangfeng Xiong
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  4. Xiuling Ji
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  7. Qi Zhang
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Correspondence to Qi Zhang.

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Li, S., Li, L., Xiong, X. et al. A novel malic enzyme gene, Mime2, from Mortierella isabellina M6-22 contributes to lipid accumulation. Biotechnol Lett 40, 1109–1118 (2018). https://doi.org/10.1007/s10529-018-2560-1

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