食品与发酵工业2024，Vol.50Issue(24)：44-50,7.DOI:10.13995/j.cnki.11-1802/ts.038659
金属β-内酰胺酶GOB-54的发现与表征
Discovery and biochemical characterization of GOB-54 metallo-β-lactamase
摘要
Abstract
This study aimed to investigate novel β-lactam resistance genes in soil and elucidate the underlying mechanisms of resist-ance,thus providing valuable clinical alerts,and insights that contribute to the development of new[3-lactam drugs.The positive clone was screened from soil metagenomic library,and resistance gene sequence was obtained by subclone construction and sequencing.Sequence a-lignment,phylogenetic tree and three-dimensional structure modeling were used to analyze the drug resistance gene.Microbroth dilution method was used to evaluate drug resistance.The gene was expressed heterologously,and the hydrolysis ability of the purified protein was tested.Sequence analysis of the subclone revealed that pZF700 contained one novel[3-lactamase gene,gob-54,exhibiting 52.3％amino acid sequence identity with the known β-lactamase genes.This gene encodes the broad-spectrum[3-lactam enzyme GOB-54.The three-di-mensional modeling of GOB-54 generated using SWISS-MODEL,revealed striking structural similarities with GOB-18 from clinical sources.The kinetic parameters indicated that GOB-54 had the highest affinity and catalytic activity for piperacillin among the β-lactam an-tibiotics tested.The Km and Kcat/Km values were found to be 121.82 μmol/L and 49.15 L/(mmol·s),respectively.In this study,the novel β-lactam resistance gene gob-54 was discovered from soil by functional metagenomics serves as a valuable reference for clinical mo-nitoring and prevention of the dissemination of β-lactam antibiotic resistance genes.关键词
β-内酰胺酶/土壤微生物/功能宏基因组学/抗生素耐药基因Key words
[3-lactamase/soil microorganism/functional metagenomics/drug resistance genes引用本文复制引用
章升霞,王绍琛,吕云斌,冯治洋..金属β-内酰胺酶GOB-54的发现与表征[J].食品与发酵工业,2024,50(24):44-50,7.基金项目
国家自然科学基金项目(32370089) （32370089）
