Fan Xiao-Man, Huang Jia-Ying, Ling Xiao-Min, Wei Wei, Su Wen-Bin, Zhang Ye-Wang
School of Pharmacy, Jiangsu University, Zhenjiang 212013, China.
Polymers (Basel). 2022 Apr 27;14(9):1770. doi: 10.3390/polym14091770.
Enzymatic preparation of low-molecular-weight chondroitin sulfate (LMWCS) has received increasing attention. In this work, a chondroitin sulfate lyase ABC (Chon-ABC) was successfully cloned, expressed, and characterized. The and of the Chon-ABC were 0.54 mM and 541.3 U mg, respectively. The maximal activity was assayed as 500.4 U mg at 37 °C in pH 8.0 phosphate buffer saline. The half-lives of the Chon-ABC were 133 d and 127 min at 4 °C and 37 °C, respectively. Enzymatic preparation of LMWCS was performed at room temperature for 30 min. The changes between the substrate and product were analyzed with mass spectrometry (MS), high-performance liquid chromatography (HPLC), gel permeation chromatography (GPC), and nuclear magnetic resonance (NMR). Overall, the Chon-ABC from is competitive in large-scale enzymatic preparation of LMWCS for its high activity, stability, and substrate specificity.
低分子量硫酸软骨素（LMWCS）的酶法制备受到了越来越多的关注。在这项工作中，成功克隆、表达并表征了硫酸软骨素裂解酶ABC（Chon-ABC）。Chon-ABC的Km和Vmax分别为0.54 mM和541.3 U mg。在37°C、pH 8.0的磷酸盐缓冲盐水中测定的最大活性为500.4 U mg。Chon-ABC在4°C和37°C下的半衰期分别为133天和127分钟。LMWCS的酶法制备在室温下进行30分钟。用质谱（MS）、高效液相色谱（HPLC）、凝胶渗透色谱（GPC）和核磁共振（NMR）分析底物和产物之间的变化。总体而言，来自[具体来源未给出]的Chon-ABC因其高活性、稳定性和底物特异性，在LMWCS的大规模酶法制备中具有竞争力。
