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Expression and Characterization of a Novel Enantioselective Lipase from Aspergillus fumigatus

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Abstract

A 1,080-bp cDNA (CGMCC 2873) encoding of a cold-active lipase of Aspergillus fumigatus (AFL67) was cloned and expressed in Escherichia coli for the first time. The new lipase, AFL67, was one-step purified by 8.30 folds through Ni–NTA affinity chromatography with a recovery of 86.8 %. The specific activity of purified AFL67 was 449 U mg−1 on p-NP hexanoate. AFL67 preferentially hydrolyzed p-nitrophenyl esters of short- and medium-chain fatty acids, with p-nitrophenyl hexanoate the maximum. The optimum temperature and pH was 15 °C and 7.5, respectively. The purified AFL67 was stable at 10–25 °C for 30 min, and in the pH range of 6.0–9.0 for 16 h (at 4 °C). Its activity was increased by 47 and 50 %, in the presence of 10 % (v/v) ethanol and isopropanol, respectively. The new lipase AFL67 highly enantioselectively deacylated (S)-α-acetoxyphenylacetic acid (APA) and o-Cl-APA, m-Cl-APA, and p-Cl-APA to (S)-mandelic acid and its derivates. These features render this cold-active novel lipase AFL67 attractive for biotechnological applications in the field of enantioselective synthesis of chiral mandelic acids, o-acylated mandelic acids, and their derivates and detergent additives.

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Acknowledgments

This research was financially supported by China National Special Fund for State Key Laboratory of Bioreactor Engineering (no. 2060204) and Shanghai Leading Academic Discipline Project (no. B505).

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People’s Republic of China

    Jiao-Jiao Shangguan, Li-qiang Fan, Xin Ju, Qing-qing Zhu, Jian Zhao & Jian-He Xu

  2. Institute of Chinese Materia Medica, Shanghai University of Chinese Traditional Medicine, Shanghai, 201203, China

    Fu-Jun Wang

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  1. Jiao-Jiao Shangguan
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  2. Li-qiang Fan
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  3. Xin Ju
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  4. Qing-qing Zhu
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Correspondence to Jian Zhao or Jian-He Xu.

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Jiao-Jiao Shangguan and Li-qiang Fan contributed equally to this work

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Shangguan, JJ., Fan, Lq., Ju, X. et al. Expression and Characterization of a Novel Enantioselective Lipase from Aspergillus fumigatus . Appl Biochem Biotechnol 168, 1820–1833 (2012). https://doi.org/10.1007/s12010-012-9899-x

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