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Heterologous expression and biochemical characterization of a cold-active lipase from Rhizopus microsporus suitable for oleate synthesis and bread making

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Abstract

Objectives

Cold-active lipases which show high specific activity at low temperatures are attractive in industrial applications in terms of product stability and energy saving. We aimed to identify novel cold-active lipase suitable for oleates synthesis and bread making.

Results

A novel lipase gene (RmLipA) from Rhizopus microsporus was cloned and heterologously expressed in Pichia pastoris. The encoding sequence displayed 75% identity to the lipase from R. niveus. The highest extracellular lipase activity of 7931 U/mL was achieved in a 5-L fermentation. The recombinant enzyme (RmLipA) was optimally active at pH 8.0 and 20–25 °C, respectively, and stable over a wide pH range of 2.0–11.0. The enzyme was a cold-active lipase, exhibiting > 80% of its maximal activity at 0 °C. RmLipA was a sn-1,3 regioselective lipase, and preferred to hydrolyze pNP esters and triglycerides with relatively long chain fatty acids. RmLipA synthesized various oleates using oleic acid and different alcohols as substrates (> 95%). Moreover, it significantly improved the quality of bread by increasing its specific volume (21.7%) and decreasing its crumb firmness (28.6%).

Conclusions

A novel cold-active lipase gene from R. microsporus was identified, and its application potentials were evaluated. RmLipA should be a potential candidate in oleates synthesis and bread making industries.

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Acknowledgements

This study was financially supported by the National Natural Science Foundation of China (No. 31871744), the Open Project Program of Beijing Key Laboratory of Flavor Chemistry, Beijing Technology and Business University (BTBU), Beijing 100048, China (SPFWHX-2016-ZD) and National Key Research and Development Program of China (2018YFD0401102).

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Author notes

  1. Man Xiang and Ling Wang have contribute equally to this work.

Authors and Affiliations

  1. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China

    Man Xiang, Ling Wang, Zhengqiang Jiang & Shaoqing Yang

  2. College of Engineering, China Agricultural University, Beijing, 100083, China

    Qiaojuan Yan

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  1. Man Xiang
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  2. Ling Wang
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  3. Qiaojuan Yan
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  4. Zhengqiang Jiang
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Correspondence to Shaoqing Yang.

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Xiang, M., Wang, L., Yan, Q. et al. Heterologous expression and biochemical characterization of a cold-active lipase from Rhizopus microsporus suitable for oleate synthesis and bread making. Biotechnol Lett 43, 1921–1932 (2021). https://doi.org/10.1007/s10529-021-03167-1

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