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

doi:10.3969/j.issn.1674-7968.2025.04.015

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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 V_max 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.

Key words： Rumen microbes Pectate lyase Alkali-tolerant Esterified Synergism

Received: 06 May 2024

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S188

Corresponding Authors: * Emirate14@zju.edu.cn;zhanghuien@zwu.edu.cn

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	BAI Shu-Ning
	LI Nuo
	DENG Qian
	LI Shi-Qi
	XU Xiao-Feng
	WANG Jia-Kun
	ZHANG Hui-En
	WANG Qian

Cite this article:

BAI Shu-Ning,LI Nuo,DENG Qian, et al. Expression and Characterization Study of an Alkaline Pectate Lyase IDSPL1-20 Derived from Rumen Microbiota[J]. 农业生物技术学报, 2025, 33(4): 867-878.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2025.04.015 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2025/V33/I4/867

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