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High-level expression and biochemical characterization of a novel cold-active lipase from Rhizomucor endophyticus

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Abstract

Objectives

To identify novel cold-active lipases from fungal sources and improve their production by heterologous expression in Pichia pastoris.

Results

A novel cold-active lipase gene (ReLipB) from Rhizomucor endophyticus was cloned. ReLipB was expressed at a high level in Pichia pastoris using high cell-density fermentation in a 5-l fermentor with the highest lipase activity of 1395 U/ml. The recombinant lipase (RelipB) was purified and biochemically characterized. ReLipB was most active at pH 7.5 and 25 °C. It was stable from pH 4.5–9.0. It exhibited broad substrate specificity towards p-nitrophenyl (pNP) esters (C2–C16) and triacylglycerols (C2–C12), showing the highest specific activities towards pNP laurate (231 U/mg) and tricaprylin (1840 U/mg), respectively. In addition, the enzyme displayed excellent stability with high concentrations of organic solvents including cyclohexane, n-hexane, n-heptane, isooctane and petroleum ester and surfactants.

Conclusions

A novel cold-active lipase from Rhizomucor endophyticus was identified, expressed at a high level and biochemically characterized. The high yield and unique enzymatic properties make this lipase of some potential for industrial applications.

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Acknowledgments

This work was financially supported by the grant of National Natural Science Foundation of China (No. 31371718), the Opening Project of Hubei Key Laboratory of Lipid Chemistry and Nutrition (201502) and the National Science Fund for Distinguished Young Scholars (No. 31325021).

Supporting information

Supplementary Table 1—Effect of organic solvents and surfactants on the activity of ReLipBa

Supplementary Figure 1—Optimal pH of the lipase from Rhizomucor endophyticus after deglycosylation.

Supplementary Figure 2—Optimal temperature (A) and thermostability (B) of the lipase from Rhizomucor endophyticus after deglycosylation.

Author information

Authors and Affiliations

  1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing, 100083, China

    Xiaojie Duan, Yu Liu, Zhengqiang Jiang & Shaoqing Yang

  2. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Hubei Key Laboratory of Lipid Chemistry and Nutrition, Wuhan, 430062, China

    Mingming Zheng

Authors
  1. Xiaojie Duan
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  2. Mingming Zheng
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  3. Yu Liu
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  4. Zhengqiang Jiang
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  5. Shaoqing Yang
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Corresponding author

Correspondence to Shaoqing Yang.

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Supplementary material 1 (DOCX 47 kb)

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Duan, X., Zheng, M., Liu, Y. et al. High-level expression and biochemical characterization of a novel cold-active lipase from Rhizomucor endophyticus . Biotechnol Lett 38, 2127–2135 (2016). https://doi.org/10.1007/s10529-016-2200-6

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  • DOI: https://doi.org/10.1007/s10529-016-2200-6

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