枯草芽孢杆菌和乳酸乳球菌对功能性重组灵芝免疫调节蛋白的胞外表达
Extracellular expression of a functional recombinant Ganoderma lucidium immunomodulatory protein by Bacillus subtilis and Lactococcus lactis.
作者信息
Yeh Chuan M, Yeh Chun K, Hsu Xun Y, Luo Qiu M, Lin Ming Y
机构信息
Department of Food Science and Biotechnology, National Chung-Hsing University, Taichung, Taiwan, Republic of China.
出版信息
Appl Environ Microbiol. 2008 Feb;74(4):1039-49. doi: 10.1128/AEM.01547-07. Epub 2007 Dec 21.
Abstract
Bacillus subtilis and Lactococcus lactis are ideal hosts for the production of extracellular heterologous proteins of major commercial importance. A recombinant gene for the novel Ganoderma lucidium immunomodulatory protein LZ-8, recombinant LZ-8, was designed encoding the same amino acid sequence but using the preferred codons for both strains and was synthesized by overlapping extension PCR. Using the signal peptide (SP) from subtilisin YaB (SP(YaB)), recombinant LZ-8 was expressed extracellularly in Bacillus subtilis and Lactococcus lactis. In the absence of SP(YaB), recombinant LZ-8 was expressed extracellularly in B. subtilis, but not in L. lactis. The three expressed recombinant LZ-8s showed different capacities for modulating the production of Th1 and Th2 cytokines by peripheral blood mononuclear cells and of tumor necrosis factor alpha by a macrophage cell line.
摘要
枯草芽孢杆菌和乳酸乳球菌是生产具有重要商业价值的细胞外异源蛋白的理想宿主。设计了一种新型灵芝免疫调节蛋白LZ-8的重组基因，即重组LZ-8，其编码相同的氨基酸序列，但使用了这两种菌株偏好的密码子，并通过重叠延伸PCR合成。利用枯草杆菌蛋白酶YaB的信号肽（SP(YaB)），重组LZ-8在枯草芽孢杆菌和乳酸乳球菌中进行细胞外表达。在没有SP(YaB)的情况下，重组LZ-8在枯草芽孢杆菌中进行细胞外表达，但在乳酸乳球菌中不表达。三种表达的重组LZ-8在调节外周血单核细胞产生Th1和Th2细胞因子以及巨噬细胞系产生肿瘤坏死因子α方面表现出不同的能力。
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