中国兽医杂志2025，Vol.61Issue(6)：1-10,10.DOI:10.20157/j.cnki.zgsyzz.2025.06.001
瘤胃产甲烷古菌噬菌体来源的mtaA的鉴定与功能分析
Identification and Functional Analysis of mtaA Derived from Rumen Methanogenic Archaea-Infecting Phages
摘要
Abstract
Methanogenic archaea play a pivotal role in the physiological homeostasis and health of ruminants.The methanogenesis process in the rumen of ruminants not only affects energy metabolism and feed utilization efficiency but is also closely related to greenhouse gas emissions.To investigate whether phages influence ruminal methanogenesis,this study assembled and analyzed the auxiliary metabolic gene(mtaA)encoded by methanogenesis-associated phages through bioinformatics analysis,prokaryotic expression,and protein purification.The methyl transfer activity of MtaA was measured using ultraviolet spectrophotometry,with coenzyme M(CoM)-dependent methylcobalamin[CH3Cob(Ⅲ)]as the substrate.Bioinformatics analysis showed that MtaA from methanogenesis-associated phages is similar to that of methanogenic archaea,with a Zn2⁺-dependent active center,suggesting its potential biological activity in methyl group transfer.Phylogenetic analysis revealed a 99.7%similarity between MtaA derived from phage and Methanobacteriaceae,indicating that the mtaA gene might have originally transferred from Methanobacteriaceae archaea to the phage genome.In this study,the recombinant plasmid pET-24a(+)-mtaA was successfully constructed,and MtaA protein was purified.Biological activity assays showed that when the concentration of MtaA protein reached 50 mg/mL,the absorbance at 520 nm significantly decreased(P=0.013).These findings demonstrate that MtaA from methanogenesis-associated phages possesses methyl transfer activity.This suggests that phages may play a role in regulating microbial communities,potentially offering a novel approach for controlling intestinal pathogens in animals,thereby contributing to animal health and food safety.关键词
瘤胃/产甲烷/噬菌体/mtaA/活性鉴定Key words
rumen/methanogenesis/phage/mtaA/activity identification分类
生物学引用本文复制引用
吴海军,李好,崔超文,李村院,刘开平,戴继鸿,倪伟,胡圣伟..瘤胃产甲烷古菌噬菌体来源的mtaA的鉴定与功能分析[J].中国兽医杂志,2025,61(6):1-10,10.基金项目
新疆自治区天山英才项目(2022TSYCCX0125) （2022TSYCCX0125）
