一株产细菌素乳酸菌的分离和鉴定

从市售3种酸乳中分离得到4株乳酸菌,通过发酵液的抑菌实验确定S1菌株为细菌素产生菌,综合S1菌株的形态学特征、生理生化特征将其初步鉴定为乳酸乳球菌(Lactococcus lactis.)。在排除过氧化氢和有机酸对指示菌的抑制作用后,确定该菌株产生的抑菌物质为细菌素,该细菌素对大肠杆菌具有较好抑制作用。     

Characterization of bacteriocin-coated antimicrobial polyethylene films by atomic force microscopy

Antimicrobial agents can be used to give antimicrobial properties to polymeric materials used to envelope foods for packaging purposes. In this study, we exploited an atomic force microscopy (AFM) analysis for the characterization of plastic films activated with antimicrobial agents. The aim was to acquire information on the distribution of the antimicrobials on plastic materials with the ultimate scope of understanding the mechanisms of interaction between antimicrobials and materials to be used for food packaging. Four polyethylene films differing in linear, EVA, and erucamide content were activated by 3 different bacteriocins as antimicrobials, namely, nisin and bacteriocins Bac162W from Lactobacillus curvatus and BacAM09 from Lactobacillus plantarum. The spectrum of activity of the bacteriocins was assayed and shown to include several strains of Listeria monocytogenes. The plastic films were activated by a previously developed coating procedure and the surfaces of the active films were examined by AFM. In addition, roughness parameters related to the single surfaces were investigated by an appropriate software. Significant differences were found between the bacteriocin activated and control (nonactivated) films and the activated surfaces showed lower values of average roughness and surface area ratio. It was not always possible to obtain a homogeneous distribution of the bacteriocin preparation following the coating procedure. This result was dependent on the bacteriocin used and its distribution on the different plastic films. Overall, the bacteriocin Bac162W showed the most homogeneous distribution while surfaces treated with nisin, showing a sort of microtexturing, always gave the highest roughness values. Although the issue needs further investigation, the connection between AFM imaging, roughness, and antimicrobial distribution on active packaging showed the potential to improve the understanding of the interactions between plastic films and antimicrobial preparations that can be important for the innovation in food packaging and science.     

双肽（IIb类）细菌素的结构特点及其与功能的关系

双肽（IIb类）细菌素通常是由革兰氏阳性菌产生的热稳定小分子（＜10 kDa）双组分抗菌肽,该类肽能够依靠特定模式基序介导肽-肽互作,形成具有跨膜能力的活性二聚体蛋白。大量研究表明,双肽细菌素具有可靠的安全性和理想的抑菌效果,在耐药菌控制方面有着巨大的应用潜力。因此,双肽细菌素发挥功能的结构特征与作用机制受到研究者们的广泛关注。本文按照双肽细菌素结构形成、肽-肽互作及肽-膜互作的逻辑梳理该类抗菌肽发挥抑菌作用的机制,同时综合现有研究阐述双肽细菌素的结构规律及影响其活性的结构要点,以期为双肽细菌素的相关研究提供新的思路。     

Class IIa bacteriocins from lactic acid bacteria: antibacterial activity and food preservation

In the last decade, a variety of ribosomally synthesized antimicrobial peptides, or bacteriocins, produced by lactic acid bacteria have been identified and characterized. As a result of these studies, insight has been gained into various fundamental aspects of biology and biochemistry such as bacteriocin processing and secretion, mechanisms of cell immunity, and structure-function relationships. In parallel, there has been a growing awareness that bacteriocins may be developed into useful antimicrobial food additives. Class IIa bacteriocins can be considered as the major subgroup of bacteriocins from lactic acid bacteria, not only because of their large number, but also because of their significant biological activities and potential applications. The present review provides an overview of the knowledge available for class IIa bacteriocins and discusses common features and recent findings concerning these substances. The activity and potential food applications of class IIa bacteriocins are a major focus of this review.     

与特定乳酸菌共培养对植物乳杆菌KLDS1.0391细菌素合成的诱导作用

植物乳杆菌KLDS1.0391与76株乳酸菌分别共培养后,测定培养物的抑菌活性和活菌数,判断与乳酸菌共培养对植物乳杆菌细菌素合成的影响及细菌素合成与菌体密度的关系。结果表明：植物乳杆菌KLDS1.0706、KLDS4.0315、KLDS4.0351、罗伊氏乳杆菌KLDS1.0736这4株菌与植物乳杆菌KLDS1.0391共培养后,抑菌活性显著增加(P＜0.01)。与罗伊氏乳杆菌共培养过程中细菌素的抑菌活性与植物乳杆菌KLDS1.0391的细胞密度呈现明显的正相关性,并且发现只有活的诱导菌与植物乳杆菌KLDS1.0391共培养才能够诱导细菌素的合成。     

响应面法优化植物乳杆菌代谢产细菌素的发酵条件

为提高一株分离自内蒙古传统发酵稀奶油“焦克”中的植物乳杆菌KLDS1.0391 代谢产细菌素量,以中性蛋白酶水解脱脂乳为培养基,以枯草芽孢杆菌为指示菌,以抑菌圈直径为考察指标,在单因素试验的基础上,采用响应面法优化发酵pH 值、温度以及接种量。结果表明：对该菌代谢产细菌素的活性影响大小依次为：发酵pH值＞接种量＞发酵温度;最优发酵条件为：pH5.1、发酵温度33℃、接种量1%。在此条件下,发酵液的抑菌圈直径为15.00mm,细菌素的效价为601.32IU/mL,较优化前提高了43.08%。在最优发酵条件下获得的实验结果与模型预测值吻合,说明所建立的模型是切实可行的。     

产细菌素戊糖乳杆菌B8发酵动力学模型的建立

从四川香肠中选育出一株具有产细菌素能力的戊糖乳杆菌B8,在10 L发酵罐内对其进行分批发酵动力学研究,建立菌体生长、产物合成、底物消耗及pH随时间变化的动力学模型,确定模型参数,得到其菌体生长动力学模型■ =0.56691-■、细菌素生产动力学模型■=75.25■ +122.94X、葡萄糖消耗动力学模型-■=5.843■+0.799 8X。pH值变化的数学模型y =■+5.992 8。结果表明,模型预测值与实验值吻合较好,为工业放大生产细菌素提供了理论依据和方法。     

Novel bacteriocins produced by Lactobacillus fermentum strains with bacteriostatic effects in milk against selected indicator microorganisms

The aim of this work was to isolate and characterize bacteriocins produced by 2 Lactobacillus fermentum strains isolated from artisanal Mexican Cocido cheese. Fractions (F ≤3 kDa) obtained from cell-free supernatants of Lb. fermentum strains J23 and J32 were further fractionated by reversed-phase HPLC on a C18 column. Antimicrobial activities of F ≤3 kDa and bacteriocin-containing fractions (BCF), obtained from fractionation of F ≤3 kDa against 4 indicator microorganisms, were determined by the disk diffusion method and growth inhibition in milk. Subsequently, isolated BCF were analyzed by reversed-phase HPLC tandem mass spectrometry. Results showed that BCF presented antimicrobial activity against the 4 indicator microorganisms tested. For J23, one of the fractions (F3) presented the highest activity against Escherichia coli and was also inhibitory against Staphylococcus aureus, Listeria innocua, Salmonella Typhimurium, and Salmonella Choleraesuis. Similarly, fractions F3 and F4 produced by J32 presented antimicrobial activity against all indicator microorganisms. Furthermore, generation time and growth rate showed that F3 from J23 presented significantly higher antimicrobial activity against the 4 indicator microorganisms (2 gram-positive and 2 gram-negative) when inoculated in milk compared with F3 from J32. Interestingly, this fraction presented a broader antimicrobial spectrum in milk than nisin (control). Reversed-phase HPLC tandem mass spectrometry analysis revealed the presence of several peptides in BCF; however, F3 from J23 that was the most active fraction of all presented only 1 bacteriocin. The chemical characterization of this bacteriocin suggested that it was a novel peptide with 10 hydrophobic AA residues in its sequence and a molecular weight of 2,056 Da. This bacteriocin and its producing strain, J23, may find application as a biopreservative against these indicator microorganisms in dairy products.     

金属离子与乳酸菌素对沙门氏菌的协同抑菌作用

通过检测唾液乳杆菌O2A发酵上清液及其所产乳酸菌素对沙门氏菌的抑菌活性,考察金属离子对乳酸菌素抑菌活性的协同作用及其机理。结果表明,单独的乳酸菌素和乳酸菌素-金属离子复配均对沙门氏菌有较好的抑菌效果;不同抑菌剂对沙门氏菌处理18 h后,细菌的生长曲线发生较大变化,且Zn2+(30 mmol/L)-乳酸菌素、Zn2+(30 mmol/L)-发酵上清液的抑菌效果较佳。扫描电子显微镜和透射电子显微镜结果表明,与标准沙门氏菌菌株相比,处理组的菌株细胞完整性均受到不同程度的破坏,破坏程度最严重的Zn2+(30 mmol/L)-乳酸菌素处理组,其抑菌机理可能是破坏细胞结构的完整性,使菌体细胞壁和细胞膜受到损伤,从而抑制细菌生长甚至杀死细菌。