Antibacterial agents specifically inhibiting lipopolysaccharide synthesis

The spread of antibiotic resistance in Gram-negative bacteria has sustained a continuing search for new agents with antibacterial activity against this important class of bacterial pathogen. Because the biosynthesis of lipopolysaccharide (LPS) is unique to Gram-negative bacteria and required by them for growth and virulence, attempts have been made to discover or design antibacterial agents acting at this site; however, no such agents have so far been developed. We now present definitive experimental data documenting design of the first member of the class of antibacterial compounds which specifically inhibit LPS synthesis. The target enzyme is 3-deoxy-D-manno-octulosonate cytidylytransferase (CMP-KDO synthetase), a cytoplasmic enzyme which activates 3-deoxy-D-manno-octulosonate (KDO) for incorporation into LPS. A specific inhibitor of CMP-KDO synthetase, alpha-C-(1,5-anhydro-7-amino-2,7-dideoxy-D-manno-heptopyranosyl)-carboxy late was designed using results of our studies of the purified enzyme. LPS synthesis ceased and lipid A precursor accumulated, causing growth stasis and perturbation of outer membrane structure and function, following delivery of the inhibitor to the intracellular target by a peptide carrier. Antibacterial action required an intact oligopeptide permease system and specific intracellular aminopeptidase activity to release inhibitor from the peptide prodrug.     

新型纳米无机抗菌剂TiO2和ZnO的广谱抗菌性研究

 选择了革兰氏阴性细菌、革兰氏阳性细菌、芽孢杆菌、酵母菌和霉菌的代表菌株,采用三角瓶振荡法及纸片扩散法,对2种新型纳米无机抗菌剂:纳米TiO₂和纳米ZnO进行了广谱抗菌性研究,并通过与日本的纳米无机抗菌剂及有机抗菌剂进行比较,结果表明2种新型纳米抗菌剂不仅对所有代表菌株表现出很好的广谱抗菌性能,而且其抑菌能力强于日本无机抗菌剂和有机抗菌剂;在此基础上对相关抗菌剂的抗菌机理进行了分析讨论.     

Human cytomegalovirus UL97 open reading frame encodes a protein that phosphorylates the antiviral nucleoside analogue ganciclovir.

Human cytomegalovirus (HCMV, a betaherpes virus) is the cause of serious disease in immunologically compromised individuals, including those with acquired immunodeficiency syndrome. One of the compounds used in the chemotherapy of HCMV infections is the nucleoside analogue 9-(1,3-dihydroxy-2-propoxymethyl)-guanine (ganciclovir). The mechanism of action of this drug is dependent on the formation of the nucleoside triphosphate, which is a strong inhibitor of the viral DNA polymerase. Thymidine kinase, which is encoded by many of the herpesviruses, catalyses the initial phosphorylation of ganciclovir. But there is no evidence for the coding of this enzyme by HCMV, and DNA sequence analysis of the HCMV genome has shown that there is no open reading frame characteristic of a herpesvirus thymidine kinase. Here we present biochemical and immunological evidence that the HCMV UL97 open reading frame codes for a protein capable of phosphorylating ganciclovir. This protein seems to be responsible for the selectivity of ganciclovir and will be useful tool in the understanding and refinement of the antiviral activity of new selective anti-HCMV compounds.     

Domain-specific recruitment of amide amino acids for protein synthesis

The formation of aminoacyl-transfer RNA is a crucial step in ensuring the accuracy of protein synthesis. Despite the central importance of this process in all living organisms, it remains unknown how archaea and some bacteria synthesize Asn-tRNA and Gln-tRNA. These amide aminoacyl-tRNAs can be formed by the direct acylation of tRNA, catalysed by asparaginyl-tRNA synthetase and glutaminyl-tRNA synthetase, respectively. A separate, indirect pathway involves the formation of mis-acylated Asp-tRNA(Asn) or Glu-tRNA(Gln), and the subsequent amidation of these amino acids while they are bound to tRNA, which is catalysed by amidotransferases. Here we show that all archaea possess an archaea-specific heterodimeric amidotransferase (encoded by gatD and gatE) for Gln-tRNA formation. However, Asn-tRNA synthesis in archaea is divergent: some archaea use asparaginyl-tRNA synthetase, whereas others use a heterotrimeric amidotransferase (encoded by the gatA, gatB and gatC genes). Because bacteria primarily use transamidation, and the eukaryal cytoplasm uses glutaminyl-tRNA synthetase, it appears that the three domains use different mechanisms for Gln-tRNA synthesis; as such, this is the only known step in protein synthesis where all three domains have diverged. Closer inspection of the two amidotransferases reveals that each of them recruited a metabolic enzyme to aid its function; this provides direct evidence for a relationship between amino-acid metabolism and protein biosynthesis.     

红霉素类抗生素分析研究进展

红霉素类抗生素药物属于广谱抗菌药,尤其对革兰氏菌特别敏感,因此其在临床及养殖业和农业中得到广泛使用。红霉素在动物体内代谢时间较长,从而会产生残留。环境中残留抗生素对人类和生物群落的影响等方面的课题已成为环境生态学家和环境分析化学家研究的热点。本文综述了近年来国内外对红霉素类抗生素的分析研究进展。     

Functional genomic analysis of phagocytosis and identification of a Drosophila receptor for E. coli

The recognition and phagocytosis of microbes by macrophages is a principal aspect of innate immunity that is conserved from insects to humans. Drosophila melanogaster has circulating macrophages that phagocytose microbes similarly to mammalian macrophages, suggesting that insect macrophages can be used as a model to study cell-mediated innate immunity. We devised a double-stranded RNA interference-based screen in macrophage-like Drosophila S2 cells, and have defined 34 gene products involved in phagocytosis. These include proteins that participate in haemocyte development, vesicle transport, actin cytoskeleton regulation and a cell surface receptor. This receptor, Peptidoglycan recognition protein LC (PGRP-LC), is involved in phagocytosis of Gram-negative but not Gram-positive bacteria. Drosophila humoral immunity also distinguishes between Gram-negative and Gram-positive bacteria through the Imd and Toll pathways, respectively; however, a receptor for the Imd pathway has not been identified. Here we show that PGRP-LC is important for antibacterial peptide synthesis induced by Escherichia coli both in vitro and in vivo. Furthermore, totem mutants, which fail to express PGRP-LC, are susceptible to Gram-negative (E. coli), but not Gram-positive, bacterial infection. Our results demonstrate that PGRP-LC is an essential component for recognition and signalling of Gram-negative bacteria. Furthermore, this functional genomic approach is likely to have applications beyond phagocytosis.     

A plant pathogen virulence factor inhibits the eukaryotic proteasome by a novel mechanism

Pathogenic bacteria often use effector molecules to increase virulence. In most cases, the mode of action of effectors remains unknown. Strains of Pseudomonas syringae pv. syringae (Pss) secrete syringolin A (SylA), a product of a mixed non-ribosomal peptide/polyketide synthetase, in planta. Here we identify SylA as a virulence factor because a SylA-negative mutant in Pss strain B728a obtained by gene disruption was markedly less virulent on its host, Phaseolus vulgaris (bean). We show that SylA irreversibly inhibits all three catalytic activities of eukaryotic proteasomes, thus adding proteasome inhibition to the repertoire of modes of action of virulence factors. The crystal structure of the yeast proteasome in complex with SylA revealed a novel mechanism of covalent binding to the catalytic subunits. Thus, SylA defines a new class of proteasome inhibitors that includes glidobactin A (GlbA), a structurally related compound from an unknown species of the order Burkholderiales, for which we demonstrate a similar proteasome inhibition mechanism. As proteasome inhibitors are a promising class of anti-tumour agents, the discovery of a novel family of inhibitory natural products, which we refer to as syrbactins, may also have implications for the development of anti-cancer drugs. Homologues of SylA and GlbA synthetase genes are found in some other pathogenic bacteria, including the human pathogen Burkholderia pseudomallei, the causative agent of melioidosis. It is thus possible that these bacteria are capable of producing proteasome inhibitors of the syrbactin class.     

Binding of a non-beta-lactam antibiotic to penicillin-binding proteins

In the search for new beta-lactam antibiotics of natural origin, the discoveries of cephamycins and sulfazecins (monobactams) were important turning points in that they accelerated many screening efforts aimed at other new compounds. In our target-directed screening for beta-lactam antibiotics using beta-lactam hypersensitive mutants, we have examined Gram-negative bacteria isolated from natural habitats and have recently reported several types of beta-lactam antibiotics such as cephabacins and formadicins. Here we report a novel antibiotic, lactivicin, found using this system. Although lactivicin has various biological activities commonly observed in beta-lactam antibiotics, it does not possess a beta-lactam ring in its molecule, but has the unique structure of a dicyclic dipeptide.     

小链霉菌HCCB10043敲除硫酯酶基因的工程菌株构建及其代谢产物的研究

小链霉菌(Streptomyces parvus)HCCB10043的主要代谢产物为脂肽类化合物A21978C,其基因组序列中包括了非核糖体肽合成酶(NRPS non ribosomal peptide synthetase)、聚酮合酶(PKS polyketide synthases)以及NRPS-PKS混合的多酶体系基因簇,它们的共同特点是在代谢产物生物合成簇中连接有一个硫酯酶域,即TE(thioesterase)domain.硫酯酶可以使已经合成的化合物链的合成过程终止,并且具有水解释放成熟脂肽以及环化线性脂肽链的功能.通过对联二吡啶类代谢产物合成簇中TE基因的敲除构得工程菌株,工程菌发酵结果表明联二吡啶类代谢产物产量减少.     

纳米TiO_2抑菌机理研究

在近荧光灯(10cm)照射下,纳米二氧化钛(TiO2)具有很强的光催化杀菌作用.作者通过TiO2对多种代表微生物的抑菌特性以及对TiO2光催化杀灭革兰氏阴、阳性细菌进行对比、常规培养验证和透射电镜观察得出结论:TiO2光催化灭菌首先是从细菌细胞壁开始,其产生的自由基能破坏细胞壁结构,使细胞壁断裂、破损,质膜解体,然后进入胞体内部破坏内膜和细胞组分,使细胞质凝聚,导致细胞内容物溢出,可出现菌体空化现象     

The Drosophila immune response against Gram-negative bacteria is mediated by a peptidoglycan recognition protein

The antimicrobial defence of Drosophila relies largely on the challenge-induced synthesis of an array of potent antimicrobial peptides by the fat body. The defence against Gram-positive bacteria and natural fungal infections is mediated by the Toll signalling pathway, whereas defence against Gram-negative bacteria is dependent on the Immune deficiency (IMD) pathway. Loss-of-function mutations in either pathway reduce the resistance to corresponding infections. The link between microbial infections and activation of these two pathways has remained elusive. The Toll pathway is activated by Gram-positive bacteria through a circulating Peptidoglycan recognition protein (PGRP-SA). PGRPs appear to be highly conserved from insects to mammals, and the Drosophila genome contains 13 members. Here we report a mutation in a gene coding for a putative transmembrane protein, PGRP-LC, which reduces survival to Gram-negative sepsis but has no effect on the response to Gram-positive bacteria or natural fungal infections. By genetic epistasis, we demonstrate that PGRP-LC acts upstream of the imd gene. The data on PGRP-SA with respect to the response to Gram-positive infections, together with the present report, indicate that the PGRP family has a principal role in sensing microbial infections in Drosophila.     

一条新的人类组氨酰tRNA合成酶类似物全长cDNA的克隆及表达谱分析

从人胎脑文库中克隆到一条长为 12 19bp的cDNA ,它编码 2 3.4ku的肽链 .该肽链与葡萄球菌组氨酰tRNA合成酶N端同源 5 8% .利用humangenomicblast可将其定位于 2 0p11.Northern杂交显示胎脑中有一约1.5kb的单一条带 .基因芯片杂交分析表明 ,其在肺癌和前列腺癌组织中表达较高 .绿荧光蛋白亚细胞定位技术显示该蛋白存在于COS7细胞的细胞质中     

新一代四环素类药物替加环素的研究进展

对于日益增多的危及生命的多药耐药菌的爆发与流行,临床上迫切需要新的抗生素,替加环素以其广谱、高效,较高的性价比,满足了人们这一需求。综述了近年来替加环素的研究进展,包括体内外的抗菌活性、抗生素后效应、药代动力学、不良反应、药物相互作用、临床研究、克服细菌耐药的机理、性价比等各个方面。替加环素是一个比较理想的药物,对大多数耐药菌都有较强的抑制力,适合于治疗复杂的具有耐药的感染。     

The Toll-like receptor 2 is recruited to macrophage phagosomes and discriminates between pathogens.

Macrophages orchestrate innate immunity by phagocytosing pathogens and coordinating inflammatory responses. Effective defence requires the host to discriminate between different pathogens. The specificity of innate immune recognition in Drosophila is mediated by the Toll family of receptors; Toll mediates anti-fungal responses, whereas 18-wheeler mediates anti-bacterial defence. A large number of Toll homologues have been identified in mammals, and Toll-like receptor 4 is critical in responses to Gram-negative bacteria. Here we show that Toll-like receptor 2 is recruited specifically to macrophage phagosomes containing yeast, and that a point mutation in the receptor abrogates inflammatory responses to yeast and Gram-positive bacteria, but not to Gram-negative bacteria. Thus, during the phagocytosis of pathogens, two classes of innate immune receptors cooperate to mediate host defence: phagocytic receptors, such as the mannose receptor, signal particle internalization, and the Toll-like receptors sample the contents of the vacuole and trigger an inflammatory response appropriate to defence against the specific organism.     

鳗鲡肝肾病病原菌的研究

从汕头地区3个养鳗场患肝肾病鳗鲡的肝脏和肾脏分离出60个菌株,用这些菌株人工感染鳗鲡能出现相同的症状,并能从肝脏或肾脏重新分离出相同的菌株,证实为该病的病原菌.这些菌株的形状基本一致,经鉴定为野生型迟呆爱德华菌(Edwardsiella tarda.wild type).这些菌株虽然是革兰氏阴性肠杆菌,但全部菌株对多粘菌素 E 高度耐药,对青霉素 G 反而较敏感.所有菌株对目前养鳗场常用的四环类抗生素及磺胺类药物高度耐药,其耐药率均达到100%,但对大多数被测试的抗菌药物敏感.     

猪胸膜放线杆菌中黏附素的研究进展

胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,APP)是引起猪传染性胸膜肺炎疾病的病原菌,APP引起猪致病的毒力因子有多种,其中黏附素作为细菌粘附宿主的第一步中的关键作用被广泛关注.黏附是APP感染宿主的第一步,分泌到革兰氏阴性菌表面的自转运黏附索调节着细菌对宿主细胞的黏附,是重要的...     

Antibacterial agents based on the cyclic D,L-alpha-peptide architecture

The rapid emergence of bacterial infections that are resistant to many drugs underscores the need for new therapeutic agents. Here we report that six- and eight-residue cyclic d,l-alpha-peptides act preferentially on Gram-positive and/or Gram-negative bacterial membranes compared to mammalian cells, increase membrane permeability, collapse transmembrane ion potentials, and cause rapid cell death. The effectiveness of this class of materials as selective antibacterial agents is highlighted by the high efficacy observed against lethal methicillin-resistant Staphylococcus aureus infections in mice. Cyclic d,l-alpha-peptides are proteolytically stable, easy to synthesize, and can be derived from a potentially vast membrane-active sequence space. The unique abiotic structure of the cyclic peptides and their quick bactericidal action may also contribute to limit temporal acquirement of drug resistant bacteria. The low molecular weight d,l-alpha-peptides offer an attractive complement to the current arsenal of naturally derived antibiotics, and hold considerable potential in combating a variety of existing and emerging infectious diseases.     

不同大豆品种对根际土壤微生物群落影响的差异

采用磷脂脂肪酸(phospholipid fatty acid, PLFA)分析法研究4个大豆品种(台湾75、中黄42、春丰早和六月半)对根际土壤微生物区系的影响。结果发现：尽管不同大豆品种在红壤(黄筋泥土)中的生物量和产量差别很大,但对根际土壤好氧菌、厌氧菌、革兰氏阴性菌、革兰氏阳性菌、放线菌和原生动物的影响不大;六月半品种显著影响根际土壤真菌PLFA总量,改变了细菌/真菌的比值;与六月半相比,春丰早显著增加了根际微生物群落的多样性、群落的优势种群和均匀度;中黄42显著提高了根际微生物群落优势种群的丰富度;中黄42和台湾75对根际土壤微生物Shannon均匀度的影响也有显著差异;但是4种大豆品种对根际土壤微生物群落的McIntosh和Gini多样性指数没有显著影响。主成分分析结果表明,六月半和中黄42品种对根际微生物的影响差异明显,而春丰早和台湾75变化相近,差异不明显。     

环二鸟苷酸对细菌运动调控的研究进展

细菌中的第二信使环二鸟苷酸(c-di-GMP)对细菌的运动性有调节作用.c-di-GMP调控鞭毛的生物合成、菌毛形成和菌毛蛋白的组成,以及其他一些与运动相关的蛋白的合成.细菌的运动性与其毒力、致病性、粘附性、趋化性、生物膜组成等密切相关.在革兰氏阴性细菌中,关于c-di-GMP的信号通路的研究较为清晰,而在革兰氏阳性细菌中,关于该信号转导通路的研究较少.此外,有关c-di-GMP的信号通路的研究主要集中在病原菌.该文主要综述了一些常见病原菌中c-di-GMP对其运动性的调控机制,为研究其他细菌c-di-GMP信号通路提供思路.     

一株光合细菌分离与初步鉴定

从西昌学院北校区养鱼池的底泥中富集分离得到一株光合细菌,该菌株厌氧培养物为红色,革兰氏染色呈阴性,繁殖方式为二均分裂。通过形态学观察及生理生化特性鉴定,鉴定为该菌株属于红假单胞菌属。