Expression and Characterization Study of an Alkaline Pectate Lyase IDSPL1-20 Derived from Rumen Microbiota
BAI Shu-Ning1, LI Nuo1, DENG Qian1, LI Shi-Qi1, XU Xiao-Feng1,2, WANG Jia-Kun1, ZHANG Hui-En2*, WANG Qian1*
1 College of Animal Sciences/Institute of Dairy Science, Zhejiang University, Hangzhou 310058, China; 2 College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China
Abstract：Pectate lyases (Pel) play critical roles in pectin degradation and hold significant biotechnological promise across diverse application in food and textile industries. In this study, a novel pectate lyase gene derived from Hu sheep (Ovis aries) rumen microbiota, IDSPL1-20 (GenBank No. PP975428), was cloned and heterologously expressed. The enzyme activity and biochemical characterizations of recombinant rIDSPL1-20 were determined. The results showed that the optimum temperature and pH of rIDSPL1-20 were 50 ℃ and pH 10.0, respectively. The rIDSPL1-20 exhibited poor thermal stability but excellent alkaline-tolerance. The residual activity maintained over 80% after treatment at pH 9.0~11.0 for 1 h. The rIDSPL1-20 was activated by the presence of 0.25~2 mmol/L Ca2+ (P<0.05). Multi-substrate specificities determination indicated that the rIDSPL1-20 showed Vmax values of (316.25±34.09) and (105.55±3.72) µmol/(min·mg) against polygalacturonic acid (PGA) and 60% esterified pectin (60% DE pectin). The rIDSPL1-20 mainly released unsaturated trigalacturonate (uG3) and unsaturated digalacturonate (uG2) from PGA and 60% DE pectin as the predominant products, indicating that the enzyme probably functions as a typical endo-pectate lyase (EC 4.2.2.2). In addition, the degradation efficiency of Cellic® CTec3 HS for peanut (Arachis hypogaea)straw was dramatically boosted by rIDSPL1-20 (P<0.05). After synergistic reaction for 2 and 4 h, the total reducing sugars yielded (0.50±0.07) and (0.57±0.14) mg/mL. This study laid the groundwork for the development of novel biomass-degrading enzymes.
白书宁, 李锘, 邓倩, 李诗棋, 徐晓锋, 王佳堃, 张慧恩, 王谦. 瘤胃微生物来源碱性果胶酸裂解酶IDSPL1-20的表达与性质研究[J]. 农业生物技术学报, 2025, 33(4): 867-878.
BAI Shu-Ning, LI Nuo, DENG Qian, LI Shi-Qi, XU Xiao-Feng, WANG Jia-Kun, ZHANG Hui-En, WANG Qian. Expression and Characterization Study of an Alkaline Pectate Lyase IDSPL1-20 Derived from Rumen Microbiota. 农业生物技术学报, 2025, 33(4): 867-878.
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