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High-speed atomic force microscopy reveals opposite traffic of processive chitinases impairs α-chitin biodegradation

文献类型： 外文期刊

作者： Qu, Mingbo 1 ; Kong, Lin 1 ; Li, Pengfei 1 ; Zhang, Aijing 1 ; Ando, Toshio 4 ; Yang, Qing 2 ;

作者机构： 1.Dalian Univ Technol, Sch Bioengn, MOE Key Lab Biointelligent Mfg, 2 Linggong Rd, Dalian 116024, Peoples R China

2.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China

3.Chinese Acad Agr Sci, Agr Genom Inst Shenzhen, Shenzhen Branch, Guangdong Lab Lingnan Modern Agr,Genome Anal Lab,M, Shenzhen 518120, Peoples R China

4.Kanazawa Univ, Nano Life Sci Inst WPI NanoLSI, Kakuma, Kanazawa 9201192, Japan

关键词： Atomic force microscope; Chitinase; Chitin; Processivity; Synergy

期刊名称：CARBOHYDRATE POLYMERS （ 影响因子：12.5； 五年影响因子：11.9 ）

ISSN： 0144-8617

年卷期： 2025 年 352 卷

页码：

收录情况： SCI

摘要： The antiparallelly organized alpha-chitin exhibits greater thermodynamic stability and is more recalcitrant to degradation than its parallel allomorph, (3-chitin, thereby impeding the efficient utilization of this natural resource. The processive chitinases usually provide the majority of catalytic potential for chitin biodegradation. Using high-speed atomic force microscopy (HS-AFM), we revealed that the opposite traffic of OfChi-h, the only processive chitinase involved in chitin biodegradation in the insect Ostrinia furnacalis, is a key factor that significantly affects alpha-chitin degradation. Compared to (3-chitin as a substrate, OfChi-h exhibited a much lower catalytic turnover and significantly reduced binding affinity to alpha-chitin. Our HS-AFM data suggested that, at the single-molecule level, OfChi-h molecules move in opposite directions along alpha-chitin fibers. Some OfChi-h molecules encounter others in a head-to-head manner, leading to their detachment from the substrate, which in turn impairs catalytic activity and processivity. Our further investigations demonstrated that the insect's strategy to avoid opposite traffic is through a synergistic interaction between OfChi-h and two non-processive endo-acting chitinases. This work reveals the mechanism by which a processive chitinase is impaired by the antiparallel organization of chitin fibers and highlights the synergism between processive and non-processive endo-acting chitinases in the degradation of recalcitrant alpha-chitin.

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