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Identification and Characterization of the Small Heat Shock Protein Hsp20 from Oenococcus oeni SD-2a

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Abstract

Oenococcus oeni can exert its function in hostile wine conditions during the malolactic fermentation process. Therefore, it is an important microbial resource for exploring resistance genes. Hsp20 is an important small heat shock protein from O. oeni. The conserved consensus motif “A-x-x-x-x-G-x-L” of Hsp20 announced its role as a member of the small heat shock protein family. The hsp20 gene from O. oeni SD-2a was cloned to create the recombinant plasmid pTriEx-Hsp20. The recombinant plasmid was transformed into Escherichia coli BL21(DE3) competent cells, and the Hsp20 protein was induced by isopropyl-β-d-thiogalactoside (IPTG). The hsp20 gene from O. oeni SD-2a was successfully expressed, and a 20-kDa fusion protein was identified by SDS-PAGE. The purified Hsp20 protein was obtained using Ni-affinity chromatography. Additionally, BL21(DE3)/Hsp20 and BL21(DE3)/Ctrl were treated at high temperatures of 42 and 52 °C, at pH values of 2.0–12.0, under oxidative shock with 0.1% (v/v) and 0.2% (v/v) H2O2, and under an osmotic shock of 430 and 860 mM NaCl to compare the effects of heterologous expression of the Hsp20 protein from O. oeni SD-2a for stress resistance. Notably, Hsp20 overexpression showed enhanced resistance than the control strain did when confronted with different elevated stress conditions. The results demonstrated heterologous expression of the hsp20 gene from O. oeni SD-2a significantly improved the resistance of the host E. coli bacteria against stress conditions.

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Funding

This funding was supported by National Natural Science Foundation of China [Grant Nos. 31560441, 31260371, 31760381] and Natural Science Foundation of Tibet [Grant Nos. XZ2017ZRG-26, XZ2018ZRG-63].

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Authors and Affiliations

  1. College of Biological and Chemical Engineering, Anhui Polytechnic University, Wuhu, 241000, Anhui, China

    Yiman Qi & Guoqiang Zhang

  2. College of Food Science and Engineering, Northwest Agriculture and Forestry University, Yangling, 712100, Shaanxi, China

    Yiman Qi, Dan Liu & Mingtao Fan

  3. Bioinformatics Center, Northwest Agriculture and Forestry University, Yangling, 712100, Shaanxi, China

    Haopeng Yu

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  1. Yiman Qi
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  2. Dan Liu
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Qi, Y., Liu, D., Yu, H. et al. Identification and Characterization of the Small Heat Shock Protein Hsp20 from Oenococcus oeni SD-2a. Curr Microbiol 77, 3595–3602 (2020). https://doi.org/10.1007/s00284-020-02168-z

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