Construction of Chitinase Complexes Using Self-Assembly Systems for Efficient Hydrolysis of Chitin.

IF 3.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS ACS Synthetic Biology Pub Date : 2024-11-20 DOI:10.1021/acssynbio.4c00613

Zhewei Shen, Yuchen Pan, Yuansheng Liu, Houhui Song, Chenggang Xu

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Abstract

Chitin biomass is the second most abundant natural polysaccharide after cellulose on the earth, yet its recalcitrance to degrade and utilize severely limits its application. However, many microorganisms, such as Serratia marcescen, can secrete a range of free chitinases to degrade chitin, though their activity is typically insufficient to meet industrial demands. In this study, we employed self-assembly systems, named SpyTag/SpyCatcher and SnoopTag/SnoopCatcher, to modularize the molecular design of CHB, ChiB, ChiC, and CBP21 derived from S. marcescens ATCC14756, and we successfully constructed a variety of chitinase complexes. The assembled complexes showed higher chitinolytic activity and stability, compared to free chitinase mixture. Moreover, the distinct arrangements and combinations of chitinases within these complexes led to varied activities, suggesting that the spatial proximity and substrate channeling effects contribute to the synergy of chitinase complexes. The findings lay a solid technical foundation for the application of chitinosome in the industrial production of N-acetylglucosamine and chitooligosaccharides.

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利用自组装系统构建几丁质酶复合物，以高效水解几丁质。

甲壳素生物质是地球上仅次于纤维素的第二大天然多糖，但其难以降解和利用的特性严重限制了它的应用。然而，许多微生物（如 Serratia marcescen）可以分泌一系列游离几丁质酶来降解几丁质，但其活性通常不足以满足工业需求。在本研究中，我们采用自组装系统（SpyTag/SpyCatcher和SnoopTag/SnoopCatcher）对来自S. marcescens ATCC14756的CHB、ChiB、ChiC和CBP21进行了模块化分子设计，并成功构建了多种几丁质酶复合物。与游离的几丁质酶混合物相比，组装后的复合物具有更高的几丁质分解活性和稳定性。此外，这些复合物中几丁质酶的不同排列和组合导致了不同的活性，表明空间接近性和底物通道效应有助于几丁质酶复合物的协同作用。这些发现为将几丁质酶体应用于 N-乙酰葡糖胺和壳寡糖的工业生产奠定了坚实的技术基础。

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来源期刊

ACS Synthetic Biology 生物-

CiteScore

8.00

自引率

10.60%

发文量

380

审稿时长

6-12 weeks

期刊介绍： The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.