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印度对虾(凡纳滨对虾)中一种c型溶菌酶的功能特性

Functional Characterization of a c-type Lysozyme from Indian Shrimp Fenneropenaeus indicus.

作者信息

Karthik Viswanathan, Kamalakannan Vijayan, Thomas Ancy, Sudheer Naduvilamuriparambu Saidumuhammed, Singh Issac S Bright, Narayanan Rangarajan Badri

机构信息

Centre for Biotechnology, Anna University, Chennai, 600025, Tamil Nadu, India,

出版信息

Probiotics Antimicrob Proteins. 2014 Jun;6(2):114-21. doi: 10.1007/s12602-013-9146-y.

Abstract

Lysozyme gene from Fenneropenaeus indicus was cloned, expressed in Escherichia coli and characterized. The cDNA consists of 477 base pairs and encodes amino acid sequence of 159 residues. F. indicus lysozyme had high identity (98%) with Fenneropenaeus merguiensis and Fenneropenaeus chinensis and exhibits low to moderate identities with lysozymes of other invertebrates and vertebrates. This lysozyme is presumed to be chicken types as it possesses two catalytic and eight cysteine residues that are conserved across c-type lysozymes and a c-terminal extension, which is a characteristic of lysozymes from marine invertebrates. Further, the antimicrobial properties of the recombinant lysozyme from F. indicus were determined in comparison with recombinant hen egg white lysozyme. This exhibited high activity against a Gram-negative pathogenic bacterium Salmonella typhimurium and two fungal strains Pichia pastoris and Saccharomyces cerevisiae in turbidimetric assay. Distribution of lysozyme gene and protein in tissues of shrimps infected with white spot syndrome virus revealed that the high levels of lysozyme are correlated with low and high viral load in abdominal muscle and tail, respectively. In conclusion, lysozyme from F. indicus has a broad spectrum of antimicrobial properties, which once again emphasizes its role in shrimp innate immune response.

摘要

克隆了印度对虾的溶菌酶基因,在大肠杆菌中进行表达并对其特性进行了研究。该cDNA由477个碱基对组成,编码159个氨基酸残基的氨基酸序列。印度对虾溶菌酶与墨吉对虾和中国对虾具有高度同源性(98%),与其他无脊椎动物和脊椎动物的溶菌酶同源性较低至中等。该溶菌酶被推测为鸡类型,因为它具有两个催化残基和八个半胱氨酸残基,这些残基在c型溶菌酶中是保守的,并且具有一个c末端延伸,这是海洋无脊椎动物溶菌酶的一个特征。此外,与重组鸡蛋清溶菌酶相比,测定了印度对虾重组溶菌酶的抗菌特性。在比浊法中,它对革兰氏阴性病原菌鼠伤寒沙门氏菌和两种真菌菌株巴斯德毕赤酵母和酿酒酵母表现出高活性。感染白斑综合征病毒的对虾组织中溶菌酶基因和蛋白质的分布表明,溶菌酶水平的升高分别与腹部肌肉和尾部低病毒载量和高病毒载量相关。总之,印度对虾溶菌酶具有广谱抗菌特性,这再次强调了其在对虾先天免疫反应中的作用。

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