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Macrobrachium rosenbergii Cu/Zn superoxide dismutase (Cu/Zn SOD) expressed in Saccharomyces cerevisiae and evaluation of the immune function to Vibrio parahaemolyticus

文献类型： 外文期刊

作者： Du, Jie 1 ; Zhu, Huanxi 3 ; Ye, Minshuo 1 ; Ma, Yan 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Agr Resources & Environm, 50 Zhongling St, Nanjing 210014, Peoples R China

2.Minist Agr, Key Lab Agroenvironm Downstream Yangtze Plain, Nanjing, Jiangsu, Peoples R China

3.Jiangsu Acad Agr Sci, Inst Anim Sci, Nanjing 210014, Peoples R China

关键词： Macrobrachium rosenbergii; Copper/zinc superoxide dismutase (Cu/ZnSOD); Saccharomyces cerevisiae; Bacterial agglutination assay; Vibrio parahaemolyticus; Immune gene expression

期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.581； 五年影响因子：4.851 ）

ISSN： 1050-4648

年卷期： 2019 年 90 卷

页码：

收录情况： SCI

摘要： Superoxide dismutases (SODs) are important antioxidant enzymes that occur in virtually all oxygen-respiring organisms, and copper/zinc SOD (Cu/ZnSOD) is one of the most important SODs. In the present study, Macrobrachiwn rosenbergii Cu/Zn-SOD was expressed in a yeast eukaryotic system. The open reading frame (ORF) of MrCu/ZnSOD was cloned into the plasmid vector pHAC181, and the recombinant plasmid was integrated into the downstream region of the GAL1 promoter in Saccharomyces cerevisiae strain GAL1-ScRCH1 via homologous recombination. The resulting recombinant MrCu/ZnSOD consisted of a 3 x HA-tag at its C-terminal. Via western blot, the molecular weight of the recombinant MrCu/ZnSOD was estimated at about 30 kDa. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of this recombinant MrCu/ZnSOD ranged from 0.556 to 0.840 mu m, and from 0.967 to 2.015 mu M, respectively. The recombinant MrCu/ZnSOD protein was able to agglutinate four Gram-negative bacterial strains, as well as two of three Gram-positive strains (except Staphylococcus aureus). This demonstrated that the recombinant protein possessed some antimicrobial activity against certain Gram-positive and Gram-negative bacteria. M. rosenbergii were fed with the recombinant yeast strain MrCu/ZnSOD for 4 weeks and then challenged with the most common crustacean pathogen, Vibrio parahaemolyticus. This group of prawns presented lower mortality, higher enzymatic activity, and higher expression of the mRNA of immune-related genes than that in the control groups. Taken together, these results suggest that MrCu/ZnSOD is an antioxidant enzyme and antimicrobial peptide involved in the crustacean innate immune system and offers protection to the host against pathogenic bacteria.

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