Identification and Characterization of the Small Heat Shock Protein Hsp20 from Oenococcus oeni SD-2a
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Identification and Characterization of the Small Heat Shock Protein Hsp20 from Oenococcus oeni SD‑2a Yiman Qi1,2 · Dan Liu2 · Haopeng Yu3 · Guoqiang Zhang1 · Mingtao Fan2 Received: 15 November 2019 / Accepted: 12 August 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
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.
Introduction Malolactic fermentation (MLF) is an important biological process in making high-quality wine. The environmental conditions for MLF are highly unfavorable with low pH and high ethanol contents. Notably, Oenococcus oeni, an acidophilic and stress-tolerant bacterium, presents a predominance against atypical conditions therein when it is used as Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00284-020-02168-z) contains supplementary material, which is available to authorized users. * Guoqiang Zhang [email protected] * Mingtao Fan [email protected] 1
College of Biological and Chemical Engineering, Anhui Polytechnic University, Wuhu 241000, Anhui, China
2
College of Food Science and Engineering, Northwest Agriculture and Forestry University, Yangling 712100, Shaanxi, China
3
Bioinformatics Center, Northwest Agriculture and Forestry University, Yangling 712100, Shaanxi, China
a starter of MLF. The potential role of O. oeni is presumed to be dependent on the promotion of stress-related proteins. Heat shock proteins are synthesized when organisms respond to environmental stress, function as molecular chaperones to safeguard proteins from denaturation and allow pro
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