用于多酶生物催化剂动态控制的可编程支架介导组装调节工具

IF 13.1 1区化学 Q1 CHEMISTRY, PHYSICAL

ACS Catalysis Pub Date : 2025-01-23 DOI:10.1021/acscatal.4c05420

Mengkai Hu, Teng Bao, Zhen Qin, Qiang Wang, Hengwei Zhang, Yujue Wang, Jiajia You, Zhenglian Xue, Rongzhen Zhang, Shang-Tian Yang, Xian Zhang, Zhiming Rao

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引用次数: 0

摘要

表面显示全细胞生物催化与传统的基于细胞内酶的方法相比具有显著的优势，但面临复杂的多酶过程和低效率的挑战。我们提出了支架介导的组装调节工具（SMART），这是一个可编程的平台，用于动态控制谷氨酸棒状杆菌表面的多酶生物催化剂。SMART将正交交联支架与双捕集器/标签对（SpyCatcher/SpyTag和SnoopCatcher/SnoopTag）集成在一起，具有可控的分子比例和高负载能力，为酶组装提供用户定义的功能。我们通过计算建模和实验验证对SMART进行了优化，并创建了双功能支架和长串联重复序列，以获得最佳的酶负载。SMART的多功能性被证明是在两个酶序列级联中共同显示，具有可调的化学计量，显著提高了麦芽糖糊精到海藻糖的转化。此外，我们整合了一个基于Bxb1重组酶的状态机（RSM）遗传电路，允许在单个细胞内对酶和支架表达进行时间调控。最后，对该体系在低成本蔗糖生产异麦芽糖中的适用性进行了验证。SMART设计代表了表面显示生物催化的重大进步，为复杂的多酶反应提供了一个可编程平台，在各种工业相关的生物催化剂中具有潜在的应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Programmable Scaffold-Mediated Assembly Regulation Tool for Dynamic Control of a Multienzyme Biocatalyst

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Programmable Scaffold-Mediated Assembly Regulation Tool for Dynamic Control of a Multienzyme Biocatalyst

Surface-displayed whole-cell biocatalysis offers significant advantages over traditional intracellular enzyme-based methods but faces challenges in complex multienzyme processes and low efficiency. We present Scaffold-Mediated Assembly Regulation Tool (SMART), a programmable platform for dynamic control of multienzyme biocatalysts on Corynebacterium glutamicum surfaces. SMART integrates orthogonal cross-linked scaffolds with the twin Catcher/Tag pairs (SpyCatcher/SpyTag and SnoopCatcher/SnoopTag), controllable molecular ratios, and high loading capacity, offering user-defined functionality for enzyme assembly. We optimized SMART through computational modeling and experimental validation and created bifunctional scaffolds and long tandem repeats for optimal enzyme loading. The versatility of SMART was demonstrated by codisplaying in a two enzyme sequential cascade with tunable stoichiometry, significantly improving the conversion of maltodextrin to trehalose. Furthermore, we integrated a Bxb1 recombinase-based state machine (RSM) genetic circuit, allowing temporal regulation of enzyme and scaffold expression within a single cell. Finally, the system’s applicability was showcased in isomaltulose production from low-cost sucrose. SMART design represents a significant advancement in surface-displayed biocatalysis, offering a programmable platform for complex multienzyme reactions with potential applications in various industrially relevant biocatalysts.

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来源期刊

ACS Catalysis CHEMISTRY, PHYSICAL-

CiteScore

20.80

自引率

6.20%

发文量

1253

审稿时长

1.5 months

期刊介绍： ACS Catalysis is an esteemed journal that publishes original research in the fields of heterogeneous catalysis, molecular catalysis, and biocatalysis. It offers broad coverage across diverse areas such as life sciences, organometallics and synthesis, photochemistry and electrochemistry, drug discovery and synthesis, materials science, environmental protection, polymer discovery and synthesis, and energy and fuels. The scope of the journal is to showcase innovative work in various aspects of catalysis. This includes new reactions and novel synthetic approaches utilizing known catalysts, the discovery or modification of new catalysts, elucidation of catalytic mechanisms through cutting-edge investigations, practical enhancements of existing processes, as well as conceptual advances in the field. Contributions to ACS Catalysis can encompass both experimental and theoretical research focused on catalytic molecules, macromolecules, and materials that exhibit catalytic turnover.