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加强原体和弹性质单体的外部合成的多维工程策略

Li-Hua Liu¹, Ying Huang², Rongrong Chen²

¹Biology Department and Institute of Marine Sciences, College of Science, Shantou University, Shantou 515063, PR China; Bio-Fermentation Research Center, Xiamen Yuanzhidao Biotechnology Co., Ltd., EYOSON Group Co., Ltd., Xiamen, 361028, PR China.

International journal of biological macromolecules

|November 19, 2025

概括

研究人员使用先进的遗传和无细胞合成方法增强了原蛋白 (CO3A1) 和弹性蛋白 (ELN) 的产生. 这一突破显著提高了工业应用的产量.

科学领域:

生物化学生物化学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

原蛋白和弹性蛋白 (ELN) 是关键的结构蛋白质,由于特定的氨基酸要求和翻译后修饰 (PTMs),它们难以外源合成.
高效的工业生产原蛋白 (例如,原蛋白III,CO3A1) 和ELN对于各种应用至关重要.

研究的目的:

开发多维策略,以改善原蛋白 (CO3A1) 和弹性蛋白 (ELN) 的外部合成.
为了提高工业规模生产的蛋白质产量.

主要方法:

优化了菌体辅助的非连续进化,以改善CO3A1基因上的T7RNA聚合酶过程性.
使用基于结构的蛋白质聚类和FACS进行增强的林氧化,选了Moumouvirus prolyl-4-hydroxylase突变体.
改善了无细胞蛋白质合成系统,以提高RNA稳定性和翻译效率.

主要成果:

鉴定了一种CO3A1基因突变 (K345E和P474L),其转录性能超过两倍.
开发了一种基4-基酶突变 (Δ2-23,S109P,L122F,V169I,P181Y) 具有三倍增加的CO3A1和ELN基化活性.
在无细胞系统中,实现了显著提高CO3A1和ELN的产量,分别达到0.31g/L和2.58g/L.

结论:

涉及转录增强,PTM优化和改善无细胞合成的多维策略显著提高了原蛋白和弹性素的生产.
这些进展对工业规模制造原蛋白和弹性纤维素有着宝贵的意义.

关键词:

无细胞蛋白质合成原蛋白和弹性纤维素定向进化是指导进化的.基因表达基因表达基因表达普罗利尔4-氧化酶是一种4-氧化酶.

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