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来自发酵氨基酸球菌的ACI-1:厌氧球菌中首个β-内酰胺酶的特性

ACI-1 from Acidaminococcus fermentans: characterization of the first beta-lactamase in Anaerobic cocci.

作者信息

Galán J C, Reig M, Navas A, Baquero F, Blázquez J

机构信息

Servicio de Microbiología, Hospital Ramón y Cajal, Instituto Nacional de Salud (INSALUD), 28034 Madrid, Spain.

出版信息

Antimicrob Agents Chemother. 2000 Nov;44(11):3144-9. doi: 10.1128/AAC.44.11.3144-3149.2000.

Abstract

Acidaminococcus fermentans belongs to the group of strictly anaerobic gram-negative cocci. All previously described Acidaminococcus strains are susceptible to beta-lactam antibiotics. An A. fermentans strain (RYC-MR95) resistant to penicillin and expanded-spectrum cephalosporin (amoxicillin and cefotaxime MICs, 64 microgram/ml) was isolated from a human perianal abscess. A fragment encoding a beta-lactamase from genomic DNA was cloned in Escherichia coli K-12 strain HB101, and the recombinant strain expressed resistance to amoxicillin (MIC, 1,024 microgram/ml) and cefotaxime (MIC, 4 microgram/ml). Clavulanic acid decreased the MICs to 8 and 0.03 microgram/ml, respectively. Analysis of the nucleotide sequence revealed a new class A beta-lactamase, ACI-1. In accordance with its biochemical properties, we propose to assign ACI-1 to functional group 2be. The ACI-1 enzyme (estimated pI 4.3) had <50% amino acid identity with any other class A beta-lactamases, the closest being ROB-1 from Haemophilus influenzae (44%). ACI-1 was closer to class A beta-lactamases from some gram-positive organisms (41 to 44% amino acid identity with Bacillus beta-lactamases) than to most class A enzymes from gram-negative organisms (TEM-1, 24.6%). The aci1 gene had a G+C content of 42.1%, in contrast with 56% G+C content for genomic DNA from A. fermentans, thus suggesting that aci1 may have been obtained by horizontal gene transfer.

摘要

发酵氨基酸球菌属于严格厌氧的革兰氏阴性球菌类。所有先前描述的发酵氨基酸球菌菌株都对β-内酰胺类抗生素敏感。从一名人类肛周脓肿患者中分离出一株对青霉素和广谱头孢菌素(阿莫西林和头孢噻肟的最低抑菌浓度为64微克/毫升)耐药的发酵氨基酸球菌菌株(RYC-MR95)。从基因组DNA中克隆出一个编码β-内酰胺酶的片段,并将其克隆到大肠杆菌K-12菌株HB101中,该重组菌株对阿莫西林(最低抑菌浓度为1024微克/毫升)和头孢噻肟(最低抑菌浓度为4微克/毫升)表现出耐药性。克拉维酸分别将最低抑菌浓度降低至8微克/毫升和0.03微克/毫升。核苷酸序列分析揭示了一种新的A类β-内酰胺酶,即ACI-1。根据其生化特性,我们建议将ACI-1归类为功能组2be。ACI-1酶(估计的等电点为4.3)与任何其他A类β-内酰胺酶的氨基酸同一性均低于50%,与流感嗜血杆菌的ROB-1最为接近(44%)。与大多数革兰氏阴性菌的A类酶(TEM-1为24.6%)相比,ACI-1与一些革兰氏阳性菌的A类β-内酰胺酶更为接近(与芽孢杆菌β-内酰胺酶的氨基酸同一性为41%至44%)。aci1基因的G+C含量为42.1%,而发酵氨基酸球菌基因组DNA的G+C含量为56%,这表明aci1可能是通过水平基因转移获得的。

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