细菌素作为生物防腐剂的研究现状

化学防腐剂潜在的安全隐患愈来愈不容忽视,寻找一种新的代用品以减少化学防腐剂所造成的潜在危害已为各国科研工作者所瞩目。自从1969年英国食品防腐剂委员会和世界卫生组织联合食品添加剂专家委员会确认nisin为食品防腐剂以来,nisin作为第一个应用于食品中的细菌素,以其生产基因稳定,抑菌范围较广的特性已陆续被许多国家接受。细菌素是某些细菌在代谢过程中通过核糖体合成机制产生的一类具有抑菌活性的多肽或前体多肽,抑菌范围不局限于同源的细菌,产生菌对其细菌素有自身免疫性［‘］能够产生细菌素的菌株很多,但并非所有的细菌素或…     

乳酸菌细菌素的研究进展

简要介绍了乳酸菌细菌素的概念和分类,并以nisin为例对其分子结构、理化性质、抑菌范围及机制、遗传控制作了较为详尽的介绍,简要概括了乳酸菌细菌素的应用和今后的研究方向。     

细菌对消毒剂抗性机理研究进展

消毒剂通过抑制膜的主动运输、抑制微生物的代谢、扰乱微生物的DNA复制、使微生物细胞裂解而导致胞内成分渗漏以及使胞内物质凝结等方面发挥灭菌作用.消毒剂在杀灭细菌和控制细菌污染方面具有重要作用,但细菌对消毒剂也会产生抗性.细菌对消毒剂的抗性机制主要通过形成生物被膜,阻挡或外排机制减少消毒剂分子进入细胞,使消毒剂分子失活,以及其他表型性耐药等4个途径来实现,其中通过形成生物被膜阻止消毒剂分子进入细菌细胞或在进入细胞前使消毒剂失去效用,在细菌对消毒剂的抗性机制中具有重要作用.以实际污染的主要菌株为对象,研究它们对常用消毒剂的抗性机制,对控制细菌污染具有极大的应用价值和现实意义.     

大肠杆菌重组乳链菌肽抗性蛋白(NSR)的表达纯化及其功能分析

乳链菌肽(Nisin)是由某些乳酸菌产生的一种阳离子抗菌肽, 而Nisin抗性蛋白(Nisin resistance protein, NSR)的表达则使一些非 Nisin产生菌获得Nisin抗性。为深入探索NSR的作用机制, 本研究在大肠杆菌中表达了去除N末端38个氨基酸残基的NSR(NSRΔ38)与GST的融合蛋白GST-NSRΔ38。通过谷胱甘肽(GSH)亲和层析和GST标签的切除后, 得到纯化的NSRΔ38, 并测定了该蛋白可能的nisin降解活性。反应产物的抑菌活性测定结果表明被NSRΔ38作用     

基因工程技术在建立噬菌体抗性机制研究中的应用

在自然发生的噬菌体抗性机制的基础上,应用基因工程技术可以建立广泛的噬菌体抗性机制,为有效解决噬菌体感染问题提供了新的策略。噬菌体编码的抗性、反义RNA技术、自杀陷阱及限制/修饰系统的应用是近年来发展起来的几种抗噬菌体策略,着重对其作用机制、研究进展及其意义作一介绍。同时指出应用基因工程技术构建的噬菌体抗性菌株应用于食品发酵工业中所存在的问题,并对其应用前景作了展望。     

羊毛硫细菌素及其应用

由基因编码、在核糖体上合成的抗菌多肽广泛分布于自然界中。人、动物、昆虫、植物和微生物都可以产生。这些抗菌多肽在食品防腐保鲜以及在药物治疗和医治肿瘤、癌症方面的潜力引起人们极大的关注［１］。近１０年来,原核生物和真核生物产生的抗菌多肽成为人们研究的热点,并取得飞速进展［１－４］。本文将主要介绍革兰氏阳性细菌产生的羊毛硫细菌素的结构、性质、生物合成,作用机制及应用。１什么叫羊毛硫细菌素由细菌基因编码、在核糖体上合成的抗菌多肽叫作细菌素。它是由某些细菌通过核糖体合成机制产生的一类具有抑菌生物活性的多肽…     

细菌抗生素和重金属协同选择抗性机制研究进展

随着各类抗生素和新型复合金属材料的不断开发和使用,环境中抗生素和重金属离子协同污染的机率不断提高,对环境选择最为敏感的细菌通过自身的进化和发展形成了二者协同选择的抗性机制,如协同抗性、交叉抗性、协同调控和生物膜诱导机制。     

细菌素及其潜在新应用的研究进展

细菌素是一种由微生物核糖体合成的抗菌肽,一般作为食品防腐剂使用。近年来,科学家挑选少数的细菌素进行深入的研究,开辟了细菌素新的研究领域,并拓宽了其应用范围。随着遗传学和纳米技术的快速发展,细菌素极有可能发展成为下一代新型抗生素、新型载体分子,肿瘤治疗的药物等。同时,科学家发现一些细菌素具有调节群体感应的功能,这一发现表明细菌素具有应用到新领域的可能。目前,革兰氏阴性菌产生的细菌素主要用于细菌素翻译修饰研究,而革兰氏阳性菌(主要是乳酸菌)产生的细菌素主要进行细菌素应用方面的研究。当前,细菌素的应用正从食品领域扩展至人类健康方面。综述了细菌素的功能及其作用效果,并且详细描述了其从食品领域到人类健康方面的应用,表明了细菌素的重要性,旨在为进一步研究细菌素在食品防腐、人类疾病防治和生物防治等领域奠定基础。     

植物病原细菌的细菌素

植物病原细菌的细菌素吴健胜,王金生（江苏南京农业大学植保系,南京２１００９５）细菌素是一种细菌的某些菌系所产生的对该种细菌的另一些菌株或关系较近的细菌有杀伤作用,非复制性的含蛋白的抗菌物质［１］。继１９４６年Ｆｒｅｄｅｒｉｃｑ报道大肠杆菌产生的大肠杆...     

细菌的噬菌体感染抗性机制

噬菌体广泛存在于生态环境中。细菌在与噬菌体长期的共进化过程中,衍化出了多种针对噬茵体感染的抗性机制。我们从宿主菌的抑制吸附、阻止噬菌体DNA注入、切断噬菌体DNA和影响其功能及流产感染等方面,对宿主菌抵抗噬菌体感染的机制进行了综述。     

Biomedical applications of nisin

Nisin is a bacteriocin produced by a group of Gram‐positive bacteria that belongs to Lactococcus and Streptococcus species. Nisin is classified as a Type A (I) lantibiotic that is synthesized from mRNA and the translated peptide contains several unusual amino acids due to post‐translational modifications. Over the past few decades, nisin has been used widely as a food biopreservative. Since then, many natural and genetically modified variants of nisin have been identified and studied for their unique antimicrobial properties. Nisin is FDA approved and generally regarded as a safe peptide with recognized potential for clinical use. Over the past two decades the application of nisin has been extended to biomedical fields. Studies have reported that nisin can prevent the growth of drug‐resistant bacterial strains, such as methicillin‐resistant Staphylococcus aureus, Streptococcus pneumoniae, Enterococci and Clostridium difficile. Nisin has now been shown to have antimicrobial activity against both Gram‐positive and Gram‐negative disease‐associated pathogens. Nisin has been reported to have anti‐biofilm properties and can work synergistically in combination with conventional therapeutic drugs. In addition, like host‐defence peptides, nisin may activate the adaptive immune response and have an immunomodulatory role. Increasing evidence indicates that nisin can influence the growth of tumours and exhibit selective cytotoxicity towards cancer cells. Collectively, the application of nisin has advanced beyond its role as a food biopreservative. Thus, this review will describe and compare studies on nisin and provide insight into its future biomedical applications.     

Nisin treatment for inactivation of Salmonella species and other gram-negative bacteria.

Nisin, produced by Lactococcus lactis subsp. lactis, has a broad spectrum of activity against gram-positive bacteria and is generally recognized as safe in the United States for use in selected pasteurized cheese spreads to control the outgrowth and toxin production of Clostridium botulinum. This study evaluated the inhibitory activity of nisin in combination with a chelating agent, disodium EDTA, against several Salmonella species and other selected gram-negative bacteria. After a 1-h exposure to 50 micrograms of nisin per ml and 20 mM disodium EDTA at 37 degrees C, a 3.2- to 6.9-log-cycle reduction in population was observed with the species tested. Treatment with disodium EDTA or nisin alone produced no significant inhibition (less than 1-log-cycle reduction) of the Salmonella and other gram-negative species tested. These results demonstrated that nisin is bactericidal to Salmonella species and that the observed inactivation can be demonstrated in other gram-negative bacteria. Applications involving the simultaneous treatment with nisin and chelating agents that alter the outer membrane may be of value in controlling food-borne salmonellae and other gram-negative bacteria.     

Nisin treatment for inactivation of Salmonella species and other gram-negative bacteria.

Nisin, produced by Lactococcus lactis subsp. lactis, has a broad spectrum of activity against gram-positive bacteria and is generally recognized as safe in the United States for use in selected pasteurized cheese spreads to control the outgrowth and toxin production of Clostridium botulinum. This study evaluated the inhibitory activity of nisin in combination with a chelating agent, disodium EDTA, against several Salmonella species and other selected gram-negative bacteria. After a 1-h exposure to 50 micrograms of nisin per ml and 20 mM disodium EDTA at 37 degrees C, a 3.2- to 6.9-log-cycle reduction in population was observed with the species tested. Treatment with disodium EDTA or nisin alone produced no significant inhibition (less than 1-log-cycle reduction) of the Salmonella and other gram-negative species tested. These results demonstrated that nisin is bactericidal to Salmonella species and that the observed inactivation can be demonstrated in other gram-negative bacteria. Applications involving the simultaneous treatment with nisin and chelating agents that alter the outer membrane may be of value in controlling food-borne salmonellae and other gram-negative bacteria.     

Immunomodulatory efficacy of nisin—a bacterial lantibiotic peptide

Nisin is a peptide bacteriocin, grouped under the category of lantibiotics. It is naturally produced by Lactococcus lactis to eliminate other competing gram‐positive bacteria from its vicinity. Moreover under certain conditions it is reported to be effective against a broad range of gram‐negative bacteria as well. Thus, it has been widely used as a safe food preservative especially in the dairy industry. Because of its wide‐scale consumption, its effect on eukaryotic cells should be of great concern. Here we examine the immunomodulatory efficacy of nisin in vitro. MTT‐based cytotoxicity assay demonstrated nisin's cytotoxicity on human T‐cell lymphoma Jurkat cells, Molt‐4 cells and freshly cultured human lymphocytes at over 200 µM concentration (IC₅₀225 µM ). The cell death mechanism induced by nisin in all these lymphocyte types was independent of oligonucleosomal DNA fragmentation, as analyzed by agarose gel electrophoresis and comet assay. Additionally, scanning electron microscope and fluorescence microscopy demonstrated the ability of nisin to activate human PMNs in vitro. Nisin‐activated neutrophils extruded intact nuclear chromatin to form NETs, well known for neutralization of virulence factors and extermination of bacterial pathogens. Nisin's presence also elevated neutrophil intracellular superoxide levels, normally produced by activated NADPH oxidase and prerequisite to NET formation. These nisin‐induced responses in cellular representatives of two separate branches of human immune system—adaptive and innate—although leading to cell death, did not include DNA fragmentation. From these findings, we propose that nisin might trigger similar AICD mechanisms in lymphocytes and neutrophils, different from conventional apoptosis which involves DNA fragmentation. Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd.     

Bioengineering nisin to overcome the nisin resistance protein

The emergence and dissemination of antibiotic resistant bacteria is a major medical challenge. Lantibiotics are highly modified bacterially produced antimicrobial peptides that have attracted considerable interest as alternatives or adjuncts to existing antibiotics. Nisin, the most widely studied and commercially exploited lantibiotic, exhibits high efficacy against many pathogens. However, some clinically relevant bacteria express highly specific membrane‐associated nisin resistance proteins. One notable example is the nisin resistance protein that acts by cleaving the peptide bond between ring E and the adjacent serine 29, resulting in a truncated peptide with significantly less activity. We utilised a complete bank of bioengineered nisin (nisin A) producers in which the serine 29 residue has been replaced with every alternative amino acid. The nisin A S29P derivative was found to be as active as nisin A against a variety of bacterial targets but, crucially, exhibited a 20‐fold increase in specific activity against a strain expressing the nisin resistance protein. Another derivative, nisin PV, exhibited similar properties but was much less prone to oxidation. This version of nisin with enhanced resistance to specific resistance mechanisms could prove useful in the fight against antibiotic resistant pathogens.     

Mechanisms of nisin resistance in Gram-positive bacteria

Nisin is the most prominent lantibiotic and is used as a food preservative due to its high potency against certain Gram-positive bacteria. However, the effectiveness of nisin is often affected by environmental factors such as pH, temperature, food composition, structure, as well as food microbiota. The development of nisin resistance has been seen among various Gram-positive bacteria. The mechanisms under the acquisition of nisin resistance are complicated and may differ among strains. This paper presents a brief review of possible mechanisms of the development of resistance to nisin among Gram-positive bacteria.     

Multimethod assessment of commercial nisin preparations

Nisin is a GRAS preservative effective against several Gram-positive organisms including Listeria monocytogenes. Commercial preparations are usually fermentation products containing 2.5% pure nisin along with insoluble material which, in this study, was found to influence the quantification and activity of nisin under different conditions. Commercially available samples of nisin were tested for efficacy using various methods, such as well diffusion, time to turbidity, and GUS (where a reporter compound is induced in response to nisin). SDS-PAGE detected a single peptide band, corresponding with the molecular weight of nisin. Protein quantified using the Bradford method indicated that the carrier of some samples was proteinaceous. Though the activity of commercially available nisin preparations is indicated on the label, end users should determine the effect of changing their source of nisin. Received 13 February 2002/ Accepted in revised form 26 July 2002     

Genetics of subtilin and nisin biosyntheses

Several peptide antibiotics have been described as potent inhibitors of bacterial growth. With respect to their biosynthesis, they can be devided into two classes: (i) those that are synthesized by a non-ribosomal mechanism and (ii) those that are ribosomally synthesized. Subtilin and nisin belong to the ribosomally synthesized peptide antibiotics. They contain the rare amino acids dehydroalanine, dehydrobutyrine, meso-lanthionine, and 3-methyl-lanthionine. They are derived from prepeptides which are post-translationally modiffied and have been termed lantibiotics because of their characteristic lanthionine bridges (Schnell et al. 1988). Nisin is the most prominent lantibiotic and is used as a food preservative due to its high potency against certain gram-positive bacteria (Mattick & Hirsch 1944, 1947; Rayman & Hurst 1984). It is produced by Lactococcus lactis strains belonging to serological group N. The potent bactericidal activities of nisin and other lantibiotics are based on depolarization of energized bacterial cytoplasmic membranes. Breakdown of the membrane potential is initiated by the formation of pores through which molecules of low molecular weight are released. A trans-negative membrane potential of 50 to 100 mV is necessary for pore formation by nisin (Ruhr & Sahl 1985; Sahl et al. 1987). Nisin occurs as a partially amphiphilic molecule (Van de Ven et al. 1991). Apart from the detergent-like effect of nisin on cytoplasmic membranes, an inhibition of murein synthesis has also been discussed as the primary effect (Reisinger et al. 1980). In several countries nisin is used to prevent the growth of clostridia in cheese and canned food. The nisin peptide structure was first described by Gross & Morall (1971), and its structural gene was isolated in 1988 (Buchman et al. 1988; Kaletta & Entian 1989). Nisin has two natural variants, nisin A and nisin Z, which differ in a single amino acid residue at position 27 (histidin in nisin A is replaced by asparagin in nisin Z (Mulders et al. 1991; De Vos et al. 1993). Subtilin is produced by Bacillus subtilis ATCC 6633. Its chemical structure was first unravelled by Gross & Kiltz (1973) and its structural gene was isolated in 1988 (Banerjee & Hansen 1988). Subtilin shares strong similarities to nisin with an identical organization of the lanthionine ring structures (Fig. 1), and both lantibiotics possess similar antibiotic activities. Due to its easy genetic analysis B. subtilis became a very suitable model organism for the identification and characterization of genes and proteins involved in lantibiotic biosynthesis. The pathway by which nisin is produced is very similar to that of subtilin, and the proteins involved share significant homologies over the entire proteins (for review see also De Vos et al. 1995b). The respective genes have been identified adjacent to the structural genes, and are organized in operon-like structures (Fig. 2). These genes are responsible for post-translational modification, transport of the modified prepeptide, proteolytic cleavage, and immunity which prevents toxic effects on the producing bacterium. In addition to this, biosynthesis of subtilin and nisin is strongly regulated by a two-component regulatory system which consists of a histidin kinase and a response regulator protein.     

乳链菌肽作用机制及其应用

乳链菌肽作为高效、安全的天然食品防腐剂已得到广泛应用。成熟的乳链菌肽含有34个氨基酸残基,由5个硫醚桥组成独特的内环结构,是研究蛋白质结构与功能的一个很好的材料。本文论述了乳链菌肽的分子结构及其与细胞膜作用的分子机制,详细阐述了乳链菌肽的作用模型。