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对l-theanine合成酶与基于PPK2的ATP再生在没有缓冲区和Mn2+优化的条件下进行比较评估

Kyungjae Yu¹, Hyun Gi Koh¹, Byung Wook Lee¹

¹Department of Biological and Chemical Engineering, Hongik University, Sejong 30016, Republic of Korea.

Enzyme and microbial technology

|February 28, 2026

概括

这项研究表明,使用工程化大肠杆菌 (E. coli) 进行有效的l-theanine生产. 本源的-氨基氨基合成酶 (GCS) 酶显示出作为生产这种功能性氨基酸的替代生物催化剂的希望.

科学领域:

生物技术是生物技术.
生物催化剂是一种生物催化剂.
代谢工程是代谢工程.

背景情况:

氨酸是一种有价值的氨基酸,具有多种营养学应用.
目前的生产主要依赖于胺甲基胺合成酶 (GMAS).
-氨基氨酸合成酶 (GCS) 对l-theanine生物合成的潜力在很大程度上尚未被探索.

研究的目的:

建立一个全细胞生物转化系统,用于生产l-氨酸.
评估本地大肠杆菌GCS (ecGCS) 作为生物催化剂的疗效.
使用多酸盐激酶2 (PPK2) 变体优化ATP再生.

主要方法:

工程化埃舍里希亚大肠杆菌菌株表达甲基菌可能GMAS (MmGMAS) 或大肠杆菌GCS (ecGCS).
集成的ATP再生系统使用聚酸激酶2 (PPK2).
高基板,没有缓冲区的全细胞转换系统.

主要成果:

在分析预测ecGCS的催化效率与MmmGMAS相比.
Rhodobacter sphaeroides PPK2 (PPK2-6) 优化了ATP再生,减少了约90%的ATP输入.
两种MmmGMAS-PPK2-6和ecGCS-PPK2-6系统都实现了高的l-theanine产量 (分别为47.9g/L和44.5g/L).

结论:

使用本地ecGCS的高效l-theanine生产在与过程相关的条件下首次被证明.
基于ecGCS的系统为GMAS生物合成提供了一个有前途的替代或补充平台.
没有缓冲区的条件有助于生产,并可能简化下游加工.

关键词:

在ATP再生过程中进行ATP再生.在L-theanine的使用中.聚酸激酶2的使用方法整细胞转化整细胞转化 γ-Glutamylcysteine合成酶的使用情况 γ-谷氨基甲基胺合成酶

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