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在Pichia pastoris中新型二次抗菌LIG的异质表达

Lu Zhao¹, Ling Li¹, Mingyang Hu¹

¹Laboratory of Molecular Nutrition and Immunity, College of Animal Science and Technology, Northeast Agricultural University, Harbin, PR China.

Journal of biotechnology

|January 5, 2024

概括

研究人员通过将两种LI结合,开发了一种二度抗菌 (AMP),LIG. 这种工程 AMP,表达在Pichia pastoris,保持强大的抗菌活性和稳定性,为AMP的生产和应用提供了一种新的方法.

科学领域:

生物化学生物化学
分子生物学分子生物学
微生物学微生物学

背景情况:

抗微生物 (AMP) 对于先天免疫非常重要.
融合LI结合了人类LL37和牛的印洛西丁,保留了抗菌特性和细胞选择性.
开发有效的AMP表达系统对于AMP的治疗应用至关重要.

研究的目的:

在Pichia pastoris中构建和表达抗微生物LI (LIG) 的二元形式.
评估复合二次 (rLIG) 的抗菌活性,溶血活性和稳定性.
利用P. pastoris表达系统,探索一种生产二维AMP的新策略.

主要方法:

使用Gly-Ser-Gly (G-S-G) 链接器结LI,形成二维LIG.
皮奇亚牧羊 (P. pastoris) 表达向量的构造 pPIC9K-6×他的-3×旗-LIG.
在P. pastoris GS115中优化复合蛋白表达,包括诱导时间,甲醇度和初始pH.
净化重组LIG (rLIG) 并评估其抗微生物和溶血活性,以及盐离子稳定性.

主要成果:

在P. pastoris GS115系统中,LIG的峰值总蛋白质表达达到189.6 mg/L.
在化酶消化后获得了5.9 mg/L的纯化重组LIG (rLIG).
rLIG表现出高的抗菌活性和低的溶血活性.
与单体LI相比,Dimeric LIG显示出可比的抗菌活性和增强的盐离子稳定性.

结论:

在P. pastoris中成功表达二元抗微生物LIG,为AMP生产提供了一种可行的方法.
迪梅里克LIG显示出强大的抗微生物疗效和稳定性,这表明治疗应用的潜力.
这项研究为优化P. pastoris系统中的AMP表达提供了有价值的见解.

关键词:

抗微生物是一种抗微生物.迪梅里克 (Dimeric) 是一种融合的表达式融合的表达式这就是Pichia pastoris.

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