Recombinant Expression of a Modified Shrimp Anti-Lipopolysaccharide Factor Gene in Pichia pastoris GS115 and Its Characteristic Analysis.
Recombinant Expression of a Modified Shrimp Anti-Lipopolysaccharide Factor Gene in Pichia pastoris GS115 and Its Characteristic Analysis.
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修饰虾抗脂多糖因子基因在毕赤酵母GS115中的重组表达及其特性分析
DOI：
10.3390/md14080152
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发表时间：
2016-08-09
期刊：
影响因子：
5.4
通讯作者：
Xiang J
中科院分区：
文献类型：
--
作者：
Yang H;Li S;Li F;Yu K;Yang F;Xiang J
具有脂多糖结合域（LBD）的抗脂多糖因子（ALFs）被认为在甲壳类动物中具有广谱抗菌活性和一定的抗病毒特性。FcALF2是从中国对虾（Fenneropenaeus chinensis）中分离到的ALFs的一个亚型。我们之前的研究表明，FcALF2修饰的LBD结构域（命名为LBDv）具有高度增强的抗菌活性。本研究设计并合成了以LBDv取代LBD的FcALF2修饰基因（mFcALF2）。该基因在酵母毕赤酵母GS115真核表达系统中成功表达，并分析了重组蛋白mFcALF2的特征。mFcALF2对革兰氏阴性菌（包括大肠杆菌、溶藻弧菌、哈威氏弧菌和副溶血性弧菌）和革兰氏阳性菌（包括地衣芽孢杆菌和表皮葡萄球菌）均有明显的抑菌活性。此外，mFcALF2可通过与病毒的预孵育，在体内抑制白斑综合征病毒（WSSV）的繁殖。本研究为水产养殖抗菌药物的开发奠定了基础。
Anti-lipopolysaccharide factors (ALFs) with a LPS-binding domain (LBD) are considered to have broad spectrum antimicrobial activities and certain antiviral properties in crustaceans. FcALF2 was one isoform of ALFs isolated from the Chinese shrimp Fenneropenaeus chinensis. Our previous study showed that a modified LBD domain (named LBDv) of FcALF2 exhibited a highly enhanced antimicrobial activity. In the present study, a modified FcALF2 gene (mFcALF2), in which the LBD was substituted by LBDv, was designed and synthesized. This gene was successfully expressed in yeast Pichia pastoris GS115 eukaryotic expression system, and the characteristics of the recombinant protein mFcALF2 were analyzed. mFcALF2 exhibited apparent antibacterial activities against Gram-negative bacteria, including Escherichia coli, Vibrio alginolyticus, Vibrio harveyi, and Vibrio parahaemolyticus, and Gram-positive bacteria, including Bacillus licheniformis and Staphylococcus epidermidis. In addition, mFcALF2 could reduce the propagation of white spot syndrome virus (WSSV) in vivo by pre-incubation with virus. The present study paves the way for developing antimicrobial drugs in aquaculture.