KSA-1,一种天然存在于肠道细菌 Kosakonia sacchari 中的 Ambler A 类广谱 β-内酰胺酶。

IF 3.7 3区 医学 Q2 INFECTIOUS DISEASES
{"title":"KSA-1,一种天然存在于肠道细菌 Kosakonia sacchari 中的 Ambler A 类广谱 β-内酰胺酶。","authors":"","doi":"10.1016/j.jgar.2024.07.008","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><p>Several bacterial species belonging to the <em>Gammaproteobacteria</em> possess intrinsic class A β-lactamase genes that may represent a source of further dissemination and acquisition to other Gram-negative species. Here we characterised KSA-1 class A β-lactamase, the gene of which was identified within the chromosome of an environmental Enterobacterales species, namely <em>Kosakonia sacchari</em>, which was also recently identified as the progenitor of an MCR-like colistin-resistance determinant.</p></div><div><h3>Methods</h3><p>In silico analysis using the GenBank database identified a class A β-lactamase gene within the chromosome of <em>K. sacchari</em> SP1 (GenBank accession no. WP_017456759). The corresponding protein KSA-1 shared 63% amino acid identity with the intrinsic CKO-1 from <em>Citrobacter koseri</em> and 53% with TEM-1. Using the <em>K. sacchari</em> DSM 100203 reference strain as a template, <em>bla</em><sub>KSA-1</sub> was amplified, cloned into the plasmid pUCp24 and expressed in <em>Escherchia coli</em> TOP10. Minimal inhibitory concentrations and kinetic parameters were obtained from the purified enzyme.</p></div><div><h3>Results</h3><p><em>K. sacchari</em> strain SP1 conferred resistance to amino-, carboxy- and ureido-penicillins only. Once produced within <em>E. coli</em>, KSA-1 showed a typical clavulanic acid-inhibited extended spectrum β-lactamase associated with a peculiar temocillin resistance profile. Kinetic assays were performed using a purified extract of KSA-1 and demonstrated a high hydrolysis rate for benzylpenicillin and piperacillin, as well as weakly extended spectrum cephalosporins. Determination of inhibitory constants showed 50% inhibitory concentration values of 2.2, 3 and 1.8 nM for clavulanic acid, tazobactam and avibactam, respectively. Analysis of sequences surrounding the <em>bla</em><sub>KSA-1</sub> gene did not reveal any mobile element that could have been involved in the acquisition of this β-lactamase gene in that species.</p></div><div><h3>Conclusion</h3><p>KSA-1 is a class A extended spectrum β-lactamase distantly related to known extended spectrum or broad-spectrum Ambler class A β-lactamases, which is highly resistant to temocillin. The <em>bla</em><sub>KSA-1</sub> gene could be considered as intrinsic within the species.</p></div>","PeriodicalId":15936,"journal":{"name":"Journal of global antimicrobial resistance","volume":null,"pages":null},"PeriodicalIF":3.7000,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2213716524001413/pdfft?md5=392f7b968cc0284a79adaa986b33316d&pid=1-s2.0-S2213716524001413-main.pdf","citationCount":"0","resultStr":"{\"title\":\"KSA-1, a naturally occurring Ambler class A extended spectrum β-lactamase from the enterobacterial species Kosakonia sacchari\",\"authors\":\"\",\"doi\":\"10.1016/j.jgar.2024.07.008\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>Several bacterial species belonging to the <em>Gammaproteobacteria</em> possess intrinsic class A β-lactamase genes that may represent a source of further dissemination and acquisition to other Gram-negative species. Here we characterised KSA-1 class A β-lactamase, the gene of which was identified within the chromosome of an environmental Enterobacterales species, namely <em>Kosakonia sacchari</em>, which was also recently identified as the progenitor of an MCR-like colistin-resistance determinant.</p></div><div><h3>Methods</h3><p>In silico analysis using the GenBank database identified a class A β-lactamase gene within the chromosome of <em>K. sacchari</em> SP1 (GenBank accession no. WP_017456759). The corresponding protein KSA-1 shared 63% amino acid identity with the intrinsic CKO-1 from <em>Citrobacter koseri</em> and 53% with TEM-1. Using the <em>K. sacchari</em> DSM 100203 reference strain as a template, <em>bla</em><sub>KSA-1</sub> was amplified, cloned into the plasmid pUCp24 and expressed in <em>Escherchia coli</em> TOP10. Minimal inhibitory concentrations and kinetic parameters were obtained from the purified enzyme.</p></div><div><h3>Results</h3><p><em>K. sacchari</em> strain SP1 conferred resistance to amino-, carboxy- and ureido-penicillins only. Once produced within <em>E. coli</em>, KSA-1 showed a typical clavulanic acid-inhibited extended spectrum β-lactamase associated with a peculiar temocillin resistance profile. Kinetic assays were performed using a purified extract of KSA-1 and demonstrated a high hydrolysis rate for benzylpenicillin and piperacillin, as well as weakly extended spectrum cephalosporins. Determination of inhibitory constants showed 50% inhibitory concentration values of 2.2, 3 and 1.8 nM for clavulanic acid, tazobactam and avibactam, respectively. Analysis of sequences surrounding the <em>bla</em><sub>KSA-1</sub> gene did not reveal any mobile element that could have been involved in the acquisition of this β-lactamase gene in that species.</p></div><div><h3>Conclusion</h3><p>KSA-1 is a class A extended spectrum β-lactamase distantly related to known extended spectrum or broad-spectrum Ambler class A β-lactamases, which is highly resistant to temocillin. The <em>bla</em><sub>KSA-1</sub> gene could be considered as intrinsic within the species.</p></div>\",\"PeriodicalId\":15936,\"journal\":{\"name\":\"Journal of global antimicrobial resistance\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-08-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2213716524001413/pdfft?md5=392f7b968cc0284a79adaa986b33316d&pid=1-s2.0-S2213716524001413-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of global antimicrobial resistance\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2213716524001413\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"INFECTIOUS DISEASES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of global antimicrobial resistance","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2213716524001413","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"INFECTIOUS DISEASES","Score":null,"Total":0}
引用次数: 0

摘要

背景:一些属于革兰氏阴性杆菌的细菌拥有固有的 A 类 β-内酰胺酶基因,这些基因可能是进一步传播和获取其他革兰氏阴性杆菌的源头。因此,我们对 KSA-1 A 类 β-内酰胺酶进行了鉴定,发现该基因存在于环境肠杆菌(即 Kosakonia sacchari)的染色体中:方法:利用 GenBank 数据库进行了内部分析,在 Kosakonia sacchari SP1(GenBank 编号:WP_065368351)的染色体中发现了一个 A 类 β-内酰胺酶基因。相应的蛋白质 KSA-1 与来自柯氏柠檬杆菌的固有 CKO-1 有 63% 的氨基酸相同性,与 TEM-1 有 53% 的氨基酸相同性。以 K. sacchari DSM 100203 参考菌株为模板,扩增 blaKSA-1,克隆到质粒 pUCp24 中,并在大肠杆菌 TOP10 中表达。从纯化的酶中获得了 MICs 和动力学参数:结果:菌株 K. sacchari SP1 只对氨基青霉素、卡博依青霉素和脲基青霉素产生抗性。一旦在大肠杆菌中产生,KSA-1就会表现出典型的克拉维酸抑制型广谱ß-内酰胺酶(ESBL),并伴有特殊的替莫西林耐药性特征。使用纯化的 KSA-1 提取物进行的动力学测定显示,该酶对苄青霉素、哌拉西林和弱广谱头孢菌素的水解率很高。抑制常数的测定显示,克拉维酸、他唑巴坦和阿维菌素的 IC50 值分别为 2.2、3 和 1.8 nM。对 blaKSA-1 基因周围序列的分析没有发现任何可能参与该物种获得这种 β-内酰胺酶基因的移动元素:结论:KSA-1 是一种 A 类 ESBL,与已知的 ESBL 关系密切,也具有很高的替莫西林活性。blaKSA-1 基因可视为该物种的固有基因。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
KSA-1, a naturally occurring Ambler class A extended spectrum β-lactamase from the enterobacterial species Kosakonia sacchari

Background

Several bacterial species belonging to the Gammaproteobacteria possess intrinsic class A β-lactamase genes that may represent a source of further dissemination and acquisition to other Gram-negative species. Here we characterised KSA-1 class A β-lactamase, the gene of which was identified within the chromosome of an environmental Enterobacterales species, namely Kosakonia sacchari, which was also recently identified as the progenitor of an MCR-like colistin-resistance determinant.

Methods

In silico analysis using the GenBank database identified a class A β-lactamase gene within the chromosome of K. sacchari SP1 (GenBank accession no. WP_017456759). The corresponding protein KSA-1 shared 63% amino acid identity with the intrinsic CKO-1 from Citrobacter koseri and 53% with TEM-1. Using the K. sacchari DSM 100203 reference strain as a template, blaKSA-1 was amplified, cloned into the plasmid pUCp24 and expressed in Escherchia coli TOP10. Minimal inhibitory concentrations and kinetic parameters were obtained from the purified enzyme.

Results

K. sacchari strain SP1 conferred resistance to amino-, carboxy- and ureido-penicillins only. Once produced within E. coli, KSA-1 showed a typical clavulanic acid-inhibited extended spectrum β-lactamase associated with a peculiar temocillin resistance profile. Kinetic assays were performed using a purified extract of KSA-1 and demonstrated a high hydrolysis rate for benzylpenicillin and piperacillin, as well as weakly extended spectrum cephalosporins. Determination of inhibitory constants showed 50% inhibitory concentration values of 2.2, 3 and 1.8 nM for clavulanic acid, tazobactam and avibactam, respectively. Analysis of sequences surrounding the blaKSA-1 gene did not reveal any mobile element that could have been involved in the acquisition of this β-lactamase gene in that species.

Conclusion

KSA-1 is a class A extended spectrum β-lactamase distantly related to known extended spectrum or broad-spectrum Ambler class A β-lactamases, which is highly resistant to temocillin. The blaKSA-1 gene could be considered as intrinsic within the species.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of global antimicrobial resistance
Journal of global antimicrobial resistance INFECTIOUS DISEASES-PHARMACOLOGY & PHARMACY
CiteScore
8.70
自引率
2.20%
发文量
285
审稿时长
34 weeks
期刊介绍: The Journal of Global Antimicrobial Resistance (JGAR) is a quarterly online journal run by an international Editorial Board that focuses on the global spread of antibiotic-resistant microbes. JGAR is a dedicated journal for all professionals working in research, health care, the environment and animal infection control, aiming to track the resistance threat worldwide and provides a single voice devoted to antimicrobial resistance (AMR). Featuring peer-reviewed and up to date research articles, reviews, short notes and hot topics JGAR covers the key topics related to antibacterial, antiviral, antifungal and antiparasitic resistance.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信