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Characterization of a novel multifunctional glycoside hydrolase family in the metagenome-assembled genomes of horse gut

文献类型： 外文期刊

作者： Hu, Lingling 1 ; Li, Xiaoyue 1 ; Li, Cunyuan 1 ; Wang, Limin 2 ; Han, Lin 1 ; Ni, Wei 1 ; Zhou, Ping 2 ; Hu, Shengwei 1 ;

作者机构： 1.Shihezi Univ, Coll Life Sci, Shihezi 832003, Xinjiang, Peoples R China

2.Xinjiang Acad Agr & Reclamat Sci, State Key Lab Sheep Genet Improvement & Hlth Prod, Urumqi 832003, Xinjiang, Peoples R China

关键词： Gut microbiota; Novel CAZymes; Gene expression

期刊名称：GENE （ 影响因子：2.6； 五年影响因子：2.7 ）

ISSN： 0378-1119

年卷期： 2024 年 927 卷

页码：

收录情况： SCI

摘要： The gut microbiota is a treasure trove of carbohydrate-active enzymes (CAZymes). To explore novel and efficient CAZymes, we analyzed the 4,142 metagenome-assembled genomes (MAGs) of the horse gut microbiota and found the MAG117.bin13 genome (Bacteroides fragilis) contains the highest number of polysaccharide utilisation loci sites (PULs), indicating its high capability for carbohydrate degradation. Bioinformatics analysis indicate that the PULs region of the MAG117.bin13 genome encodes many hypothetical proteins, which are important sources for exploring novel CAZymes. Interestingly, we discovered a hypothetical protein (595 amino acids). This protein exhibits potential CAZymes activity and has a lower similarity to CAZymes, we named it BfLac2275. We purified the protein using prokaryotic expression technology and studied its enzymatic function. The hydrolysis experiment of the polysaccharide substrate showed that the BfLac2275 protein has the ability to degrade alpha-lactose (156.94 U/mg), maltose (92.59 U/mg), raffinose (86.81 U/mg), and hyaluronic acid (5.71 U/mg). The enzyme activity is optimal at pH 5.0 and 30 degree celsius, indicating that the hypothetical protein BfLac2275 is a novel and multifunctional CAZymes in the glycoside hydrolases (GHs). These properties indicate that BfLac2275 has broad application prospects in many fields such as plant polysaccharide decomposition, food industry, animal feed additives and enzyme preparations. This study not only serves as a reference for exploring novel CAZymes encoded by gut microbiota but also provides an example for further studying the functional annotation of hypothetical genes in metagenomic assembly genomes.

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