Expression and characterization of a novel microbial GH9 glucanase, IDSGLUC9-4, isolated from sheep rumen

文献类型： 外文期刊

第一作者： Zhu, Yongzhen

作者： Zhu, Yongzhen;Wang, Kaiying;Zhang, Tietao;Zhu, Yongzhen;Bai, Shuning;Li, Nuo;Wang, Jun-Hong;Wang, Jia-Kun;Wang, Qian;Zhu, Yongzhen;Bai, Shuning;Li, Nuo;Wang, Jun-Hong;Wang, Jia-Kun;Wang, Qian

作者机构：

关键词： Expression; beta-Glucan; Glycoside Hydrolase; Hydrolysis; Rumen Microbiome

期刊名称：ANIMAL BIOSCIENCE （ 影响因子：2.5； 五年影响因子：2.8 ）

ISSN： 2765-0189

年卷期： 2024 年 37 卷 9 期

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收录情况： SCI

摘要： Objective: This study aimed to identify and characterize a novel endo-(3-glucanase, IDSGLUC9-4, from the rumen metatranscriptome of Hu sheep. Methods: A novel endo-(3-glucanase, IDSGLUC9-4, was heterologously expressed in Escherichia coli and biochemically characterized. The optimal temperature and pH of recombinant IDSGLUC9-4 were determined. Subsequently, substrate specificity of the enzyme was assessed using mixed-linked glucans including barley (3-glucan and Icelandic moss lichenan. Thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), matrix assisted laser desorption ionization time of flight mass spectrometry analyses were conducted to determine the products released from polysaccharides and cello-oligosaccharides substrates. Results: The recombinant IDSGLUC9-4 exhibited temperature and pH optima of 40 degrees C and pH 6.0, respectively. It exclusively hydrolyzed mixed-linked glucans, with significant activity observed for barley (3-glucan (109.59 +/- 3.61 mu mol/mg min) and Icelandic moss lichenan (35.35 +/- 1.55 mu mol/mg min). TLC and HPLC analyses revealed that IDSGLUC9-4 primarily released cellobiose, cellotriose, and cellotetraose from polysaccharide substrates. Furthermore, after 48 h of reaction, IDSGLUC9-4 removed most of the glucose, indicating transglycosylation activity alongside its endo-glucanase activity. Conclusion: The recombinant IDSGLUC9-4 was a relatively acid-resistant, mesophilic endo-glucanase (EC 3.2.1.4) that hydrolyzed glucan-like substrates, generating predominantly G3 and G4 oligosaccharides, and which appeared to have glycosylation activity. These findings provided insights into the substrate specificity and product profiles of rumen-derived GH9 glucanases and contributed to the expanding knowledge of cellulolytic enzymes and novel herbivore rumen enzymes in general.

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