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一个由毒素-抗毒素驱动的生物传感器平台,用于对d-氨酸3-胺酶的定向进化

Yuying Han¹, Jiaqi Li¹, Yaxin He¹

¹Key Laboratory of Industrial Fermentation Microbiology of the Ministry of Education; Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology; National Engineering Laboratory for Industrial Enzymes, Tianjin 300457, P. R. China.

Journal of agricultural and food chemistry

|February 25, 2026

概括

科学家们设计了一个新的选平台,用于酶进化. 这种基于毒素-抗毒素的系统 (TASP) 提高了酶活性和稳定性,克服了以前方法的局限性.

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?
合成生物学合成生物学

背景情况:

生物传感器辅助查对于酶导向进化至关重要.
现有的基于抗生素耐药性的方法面临着选择压力的下降.
对于高通量选,需要一个强大的,可持续的选择压力.

研究的目的:

开发一种新的基于毒素-抗毒素的高通量查平台 (TASP).
通过使用TASP系统进行d-粉-3-酶 (DAE) 工程.
通过定向进化来增强酶催化活性和稳定性.

主要方法:

开发了一种纳入毒素-抗毒素系统 (AtaT-AtaR) 的TASP,用于细胞自主选择压力.
设计了一种对d-素敏感的生物传感器 (DB),配备了增强的pPsiA促进剂,以获得更广泛的动态范围和更好的信号噪声比.
利用mCherry作为记者基因,将DAE活动与细胞生长表型联系起来,用于视觉和定量分析.

主要成果:

设计的ppsiA促进剂实现了150mM的d-allulose动态范围和14.2倍的信号对噪声增强.
该TASP平台成功将DAE催化活性与细胞生长联系起来.
确定了一种DAE突变 (M4-2),其催化活性增加了5.2倍,半衰期在60°C时长了3.5倍.

结论:

建立了一种可靠的TASP,用于对DAE变种进行高通量选.
推进了对酶工程的生长合选择策略的开发.
该TASP平台提供了一个可持续和有效的方法,用于定向的酶进化.

关键词:

在d-allulose 3-epimerase中使用.增长合生物传感器高通量选的高通量选毒素-抗毒素系统的毒素.

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