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来自d-果糖的ATP再生的无细胞反应系统

Franziska Kraußer¹, Kenny Rabe¹, Christopher M Topham²

¹Chair of Bioprocess Engineering, Institute of Natural Materials Technology, TU Dresden, Bergstraße 120, 01062 Dresden, Germany.

ACS synthetic biology

|March 27, 2025

概括

这项研究介绍了使用d-果糖的无细胞腺三酸盐 (ATP) 再生系统. 优化的酶可以实现高ATP产量,使工业生物工艺更加可行.

科学领域:

生物技术是生物技术.
酶工程是什么? 酶工程是什么?
代谢工程是代谢工程.

背景情况:

依赖亚丁三酸盐 (ATP) 的体外生物过程对于工业应用,如无细胞蛋白质合成至关重要.
高昂的ATP成本是这些生物工艺广泛实施的一个主要限制.
开发高效,经济的ATP再生系统对于推进这些技术至关重要.

研究的目的:

开发和演示一种使用低成本基质的新型无细胞ATP再生系统.
为提高ATP生产效率,设计了一种改进的光基托拉酶酶.
为了优化整体酶级联以最大限度地提高ATP产量和生产力.

主要方法:

设计了一个无细胞系统,利用从d-果糖和无机酸盐中产生的酸乙烯.
采用半理性工程方法来优化Bifidobacterium adolescentis phosphoketolase (Bad.F6Pkt) 的使用.
集成工程Bad.F6Pkt,乙酸激酶,甘激酶和l-rhamnose异构酶成为一个完整的ATP再生级联.

主要成果:

改造的Bad.F6Pkt变种H548N显示d-果糖,d-红糖和糖类甲的活性显著增加.
从d-果糖中证明了ATP再生,其固态度为每molC6基1molATP.
整个系统的ATP产量为2.53mol ATP/mol d-果糖,生产率为7.2mM/h.

结论:

开发的无细胞ATP再生系统有效地利用了低成本的d-果糖.
基酶的酶工程对于提高ATP生产效率至关重要.
该系统为工业ATP依赖生物工艺提供了具有成本效益的解决方案.

关键词:

在ATP辅助因子的再生过程中,乙烯酸酸盐的合成方法在体外生物催化剂.分子建模分子建模光基托拉酶是一种光基托拉酶.半导体酶工程半导体酶工程

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