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Effects of GSH1 and GSH2 Gene Mutation on Glutathione Synthetases Activity of Saccharomyces cerevisiae

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Abstract

In this paper, three mutants from wild Saccharomyces cerevisiae HBU2.558, called U2.558, UN2.558, and UNA2.558, were screened by UV, sodium nitrite, Atmospheric and room temperature plasma, respectively. Glutathione production of the three mutants increased by 41.86, 72.09 and 56.76%, respectively. We detected the activity of glutathione synthetases and found that its activity was improved. Amino acid sequences of three mutant colonies were compared with HBU2.558. Four mutants: Leu51→Pro51 (L51P), Glu62→Val62 (E62V), Ala332→Glu332 (A332E) and Ser653→Gly653 (S653G) were found in the analysis of γ-glutamylcysteine ligase. L51 is located adjacently to the two active sites of GCL/E/Mg2+/ADP complex in the overall GCL structure. L51P mutant spread distortion on the β-sheet due to the fact that the φ was changed from −50.4° to −40.2°. A mutant Leu54→Pro54 (L54P) was found in the analysis of glutathione synthetase, and L54 was an amino acid located between an α-helix and a β-sheet. The results confirm that introduction of proline located at the middle of the β-sheet or at the N- or C-terminal between α-helix and β-sheet or, i.e., L51P and L54P, changed the φ, rigidity, hydrophobicity and conformational entropy, thus increased protein stability and improved the enzyme activity.

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Abbreviations

S .cerevisiae :

Saccharomyces cerevisiae

GSH:

Glutathione

ARTP:

Room temperature plasma

GCL:

γ-Glutamylcysteine ligase

GSS:

Glutathione synthetase

YPD:

Yeast extract-peptone-dextrose

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Acknowledgements

The work was supported by the Science Research Plan of Hebei Higher Schools (No. Z2010225), open fund of Key laboratory (No. 3333112) and Bioengineering key discipline of Hebei Province.

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Authors and Affiliations

  1. Engineering Laboratory of Microbial Breeding and Preservation of Hebei Province, Key Laboratory of Microbial Diversity Research and Application of Hebei Province, Key Discipline of Biological Engineering of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China

    Wen Xu, Haiyan Jia, Longmei Zhang, Haiyan Wang, Hui Tang & Liping Zhang

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  1. Wen Xu
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  2. Haiyan Jia
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  3. Longmei Zhang
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Correspondence to Liping Zhang.

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Xu, W., Jia, H., Zhang, L. et al. Effects of GSH1 and GSH2 Gene Mutation on Glutathione Synthetases Activity of Saccharomyces cerevisiae . Protein J 36, 270–277 (2017). https://doi.org/10.1007/s10930-017-9731-0

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