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A novel β-glucosidase with lipolytic activity from a soil metagenome

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Abstract

Moonlighting proteins have two different functions within a single polypeptide chain. Exploring moonlighting enzymes from the environment using the metagenomic approach is interesting. In the present study, a novel β-glucosidase gene, designated as bgl1D, with lipolytic activity (renamed Lip1C) was cloned through function-based screening of a metagenomic library from uncultured soil microorganisms. The deduced amino acid sequence comparison and phylogenetic analysis also indicated that Lip1C and other putative lipases are closely related. Biochemical characterization demonstrated that the maximum activity of the recombinant Lip1C protein occurs at pH 8.0 and 30°C using 4-nitrophenyl butyrate as substrate. The putative lipase had an apparent K m value of 0.88 mmol/L, a k cat value of 212/min, and a k cat/K m value of 241 L/mmol/min. Lip1C exhibited habitat-specific characteristics with 5 mmol/L AlCl3, CuCl2, and LiCl. The characterization of the biochemical properties of Lip1C enhances our understanding of this novel moonlighting enzyme isolated from a soil metagenome.

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Abbreviations

E. coli :

Escherichia coli

PCR:

Polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

Ni-NTA:

Nickel-nitrilotriacetic acid

EDTA:

Ethylenediamine tetraacetic acid

SDS:

Sodium dodecyl sulfate

ORF:

Open reading frame

pNPB:

4-Nitrophenyl butyrate

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Acknowledgments

This research was supported in part by the Ph.D. Programs Foundation of Ministry of Education of China (Grant No. 200805930002), the Natural Science Foundation of Guangxi Zhuang Autonomous Region of China (Grant No. 2010GXNSFB013019, 2011GXNSFA018073).

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Authors and Affiliations

  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, and College of Life Science and Technology, Guangxi University, 100 Daxue East Road, Nanning, Guangxi, 530004, People’s Republic of China

    Cheng-Jian Jiang, Gao Chen, Jie Huang, Ke Jin, Pei-Hong Shen & Jun-Fang Li

  2. College of Chemistry and Ecology Engineering, Guangxi University for Nationalities, 188 Daxue East Road, Nanning, Guangxi, 530006, People’s Republic of China

    Qin Huang & Bo Wu

Authors
  1. Cheng-Jian Jiang
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  2. Gao Chen
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  3. Jie Huang
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  4. Qin Huang
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  5. Ke Jin
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  6. Pei-Hong Shen
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  7. Jun-Fang Li
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  8. Bo Wu
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Corresponding author

Correspondence to Bo Wu.

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Supplementary Fig. 1S

SDS-PAGE analysis of recombinant proteins of Lip1C. Proteins were separated by 12% SDS-PAGE and then stained with Coomassie brilliant blue G-250. A and B: Lane 1 protein molecular weight markers; Lane 2 total protein of E. coli Tuner (DE3) pLacI harboring empty pETBlue-2 as control; Lane 3 total protein of E. coli Tuner (DE3) pLacI harboring the recombinant lip1C in pETBlue-2 induced by addition of 0.5 mM IPTG; Lane 4 sample purified by the nickel-nitrilotriacetic acid column method. The recombinant Lip1C protein is indicated by the black arrow (DOC 2464 kb)

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Jiang, CJ., Chen, G., Huang, J. et al. A novel β-glucosidase with lipolytic activity from a soil metagenome. Folia Microbiol 56, 563–570 (2011). https://doi.org/10.1007/s12223-011-0083-4

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  • DOI: https://doi.org/10.1007/s12223-011-0083-4

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