氨基酸的碳负合成使用基于细胞的自由生物催化剂
Shaafique Chowdhury1, Ray Westenberg1,2, Kimberly Wennerholm3
1School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, Atlanta, Georgia 30332, United States.
ACS synthetic biology
|November 21, 2024
概括
研究人员开发了一种无细胞生物催化剂,用于从二氧化碳等价物中合成负碳氨基酸. 该系统有效地产生甘氨酸和氨酸,克服了工业应用传统生物方法的局限性.
科学领域:
- 合成生物学和代谢工程.
- 生物催化和酶工程.
- 碳捕获和利用 (CCU).碳捕获和利用.
背景情况:
- 生物系统可以将二氧化碳 (CO2) 转化为化学物质,但天然的二氧化碳固定速度太慢,无法用于工业用途.
- 有机体中工程代谢途径具有局限性,包括缓慢的碳固定和生长和合成之间的碳分配.
- 以前使用工程生物固定二氧化碳的尝试导致了细胞生长,但没有化学合成.
研究的目的:
- 设计一种基于细胞自由表达 (CFE) 的多酶生物催化剂,用于从二氧化碳等效物中碳负合成甘氨酸和血清.
- 为了克服在全细胞系统中看到的缓慢的二氧化碳固定率和低效的产品合成的局限性.
- 建立一个新的生物催化平台,从二氧化碳中生产有价值的化学品.
主要方法:
- 开发了一种基于溶酸盐的细胞自由表达 (CFE) 系统,使用多酶生物催化剂.
- 纳入的四基酸盐 (THF) 依赖的形式固定,还原性甘氨酸合成和血清合成途径.
- 实现了依赖酸盐脱酶的NAD(P) H再生,以驱动反应和优化基因比率和辅因子循环.
主要成果:
- 使用基于CFE的生物催化剂,达到30%的甲酸盐转化为血清素和甘氨酸,超过之前净化的酶系统效率 (22%).
- 即使经过200倍稀释,也证明了生物催化剂活性,允许更高的基质负载而不会增加细胞溶解物成本.
- 确定NAD(P) H再生对于驱动接近热力学平衡的反应至关重要,以及高效的THF循环利用降低了辅助因子成本.
结论:
- 本研究介绍了首次使用基于CFE的生物催化剂合成氨基酸,该生物催化剂捕获碳负生产的二氧化碳等价物.
- 与全细胞或纯化酶系统相比,CFE系统在效率,基板负载和成本效益方面具有优势.
- CFE平台具有扩展到其他有价值的化学物质 (如酸盐) 的潜力,使CO2化学合成中的更广泛应用成为可能.
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