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在益生菌中构建无抗生素蛋白表达系统用于卵蛋白生物合成 Escherichia coli Nissle 1917

Cheng Liu^1,2,3,4,5, Xueqin Lv^1,2,3,4, Long Liu^1,2,3,4

¹Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.

Journal of agricultural and food chemistry

|April 4, 2024

概括

使用工程 *Escherichia coli* Nissle 1917 生产的细胞培养卵蛋白 (OVA) 为蛋提供了一个环保的替代品. 这项研究在一种新的,无抗生素的表达系统中实现了创纪录的高OVA产量.

科学领域:

生物技术是生物技术.
微生物工程微生物工程
蛋白质表达方式

背景情况:

卵蛋白 (OVA) 是主要的蛋蛋白,但其生产对环境产生重大影响.
细胞培养OVA是一种可持续的替代品,可以减少全球变暖和土地利用.
益生菌 *Escherichia coli* Nissle 1917 (EcN) 提供了一个强大的,无抗生素的宿主,用于异质蛋白质的生产,因为它的内源性质粒.

研究的目的:

开发一种高效的,无抗生素的系统,用于*Escherichia coli*Nissle 1917 (EcN) 中异质卵蛋白 (OVA) 的生产.
通过基因工程优化OVA产量,包括促进体和核糖体结合部位的修改.
描述重组OVA的生物化学特性,并将其与天然OVA进行比较.

主要方法:

在内源性等离子体上使用OVA表达带进行工程EcN.
在最初的OVA表达式中使用了战术促进器 (P_tac).
实现了双促进体系统 (P_P5 + P_tac) 和替代的核糖体结合位 (RBS) 增强表达.
在3L生物反应器中扩大生产规模.

主要成果:

在摇瓶中实现了20 mg·L-1的初始OVA产量.
优化表达增加了30毫克·L-1和42毫克·L-1的产量,使用双促进剂和RBS替代.
在使用优化 EcN 菌株 (EcNP-P28) 和等离子体 (L28) 的生物反应器中达到创纪录的 3.70 g·L-1 OVA.
再组合OVA表现出与天然OVA相似的生化特征.

结论:

在EcN中成功建立了高产,无抗生素的异质蛋白表达系统.
证明了工程EcN在可持续大规模生产OVA等重组蛋白质方面的潜力.
开发的系统代表了微生物蛋白质生产的重大进步,实现了 prokaryotes 中报告的最高 OVA 产量.

关键词:

大肠杆菌 Escherichia coli 尼斯尔 1917 年没有抗生素的蛋白质表达系统.优化基因表达的优化卵性白蛋白 (Ovalbumin) 是一种卵性白蛋白.

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