数字农科院2.0

Characterization of a novel chondroitinase and its lab-scale performance in chondroitin sulfate degradation from meat processing wastewater

文献类型： 外文期刊

作者： Ruiyun Wu;Dequan Zhang;Mengyuan Xiao;Qian Shen;Linggao Liu;Roshani Dumila;Pinglan Li;Zhenyu Wang;Nan Shang

作者机构：

关键词： Catalytic mechanism;Chondroitinase;Enzymatic activity;Substrate specificity;Wastewater treatment

期刊名称： Journal of Cleaner Production

ISSN： 0959-6526

年卷期： 2025 年 531 卷

页码：

收录情况： SCIE（2025版） ; ; EI（2025版）

摘要： Meat processing wastewater presents a significant environmental challenge, largely due to inefficient enzymatic degradation of chondroitin sulfate (CS). This study introduces a novel chondroitinase (Ps_Chase) from Pseudarthrobacter sp. PL-410, exhibiting a 50 % higher enzymatic activity (850.7 U/mg) than commercial Chase ABC. Ps_Chase displays exceptional thermostability, maintaining over 80 % activity after 180 min at 37 °C and demonstrating half-lives of 420 min (CS-A) and 480 min (CS-C). It shows high specificity for CS-C, with a low Km (0.0425 mg/mL) and high Vmax (0.171 μmol/min·mL), and achieves a 59.36 % yield of 1–3 kDa oligosaccharides, increasing bioavailability by 140 %. Compared to other enzymes, Ps_Chase reduced chitosan molecular weight by 48.8 % and increased absorbance at 232 nm by 1.5-2-fold, demonstrating superior depolymerization efficiency. Structural analysis highlights several conserved catalytic residues (H271, R334, Y280, E445), with mutagenesis confirming their essential roles in catalysis. Molecular simulations revealed stable substrate binding, enhanced compactness, and the highest hydrogen bond count (9.68) for CS-C. QM/MM simulations identified a two-step proton transfer mechanism, with H271 and R334 catalyzing cleavage via β-elimination, exhibiting a rate-limiting energy barrier of 26.11 kcal/mol, which was significantly lower than for CS-B (37.89 kcal/mol). Y280 was found to stabilize the carbanion intermediate through hydrogen bonding. In volume-dependent experiments (0.5–3 L), Ps_Chase consistently reduced BOD and COD levels by up to 42 %. This outcome indicates the effective depolymerization of CS-rich substrates under controlled conditions. These results demonstrate Ps_Chase's catalytic efficiency, substrate selectivity, and mechanistic features, providing a basis for future exploration of environmentally relevant applications in chondroitin sulfate valorization.

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