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Molecular Biotechnology

, Volume 61, Issue 12, pp 892–904 | Cite as

A Novel VIII Carboxylesterase with High Hydrolytic Activity Against Ampicillin from a Soil Metagenomic Library

  • Fang Nan
  • Junwei Jiang
  • Shenglu Wu
  • Yueqi Zhang
  • Jiarong Qiu
  • Beibei Qiao
  • Shan Li
  • Zhihong XinEmail author
Review

Abstract

A novel carboxylesterase gene, named dlfae4, was discovered and sequenced from a soil metagenomic library. The dlfae4 gene was composed of 1017 base pairs encoding 338 amino acid residues with a predicted molecular mass of 37.2 kDa. DLFae4 exhibited strong hydrolytic activity towards methyl ferulate under optimum pH and temperature conditions (pH 8.6, 50 °C) and displayed remarkable thermostability, with residual activity as high as 50% after incubation for 3 h at 60 °C. A family VIII esterase DLFae4 was found to contain a typical serine residue within the S-X-X-K motif, which serves as a catalytic nucleophile in class C β-lactamases and family VIII esterases. As a consequence of its high sequence similarity with β-lactamases, DLFae4 exhibited significant hydrolytic activity towards ampicillin. In addition, DLFae4 was found to be the first known member of family VIII carboxylesterases with phthalate-degrading ability. Site-directed mutagenesis studies revealed that Ser11, Lys14, and Tyr121 residues play an essential catalytic role in DLFae4. These new findings, which are of great importance for further in-depth research and engineering development of carboxylesterases, should advance the implementation of biotechnological applications.

Keywords

Carboxylesterase Metagenomics Hydrolytic ampicillin Phthalate degradation Thermostability 

Notes

Acknowledgements

This research was supported by Fundamental Research Funds for the Central Universities (KYYJ201708) and by Six Talent Peaks Project in Jiangsu Province and by special funds of agro-product quality safety risk assessment of Ministry of Agriculture of the People’s Republic of China (GJFP201801104).

Compliance with Ethical Standards

Conflict of interest

The authors declare that there is no conflict of interest.

Human and Animal Rights

This article does not contain any studies with human or animal subjects.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Fang Nan
    • 1
  • Junwei Jiang
    • 1
  • Shenglu Wu
    • 1
  • Yueqi Zhang
    • 1
  • Jiarong Qiu
    • 1
  • Beibei Qiao
    • 1
  • Shan Li
    • 1
  • Zhihong Xin
    • 1
    Email author
  1. 1.Key Laboratory of Food Processing and Quality Control, College of Food Science and TechnologyNanjing Agricultural UniversityNanjingPeople’s Republic of China

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