Nisin expression production from Lactococcus lactis in milk whey medium

BACKGROUND: Nisin is a commercially available bacteriocin produced by Lactococcus lactis ATCC 11454 and used as a natural agent in the biopreservation of food. In the current investigation, milk whey, a byproduct from dairy industries was used as a fermentation substrate for the production of nisin. Lactococcus lactis ATCC 11454 was developed in a rotary shaker (30 °C/36 h/100 rpm) using two different media with milk whey (i) without filtration, pH 6.8, adjusted with NaOH 2 mol L^?1 and without pH adjustment, both autoclaved at 121 °C for 30 min, and (ii) filtrated (1.20 µm and 0.22 µm membrane filter). These cultures were transferred five times using 5 mL aliquots of broth culture for every new volume of the respective media. RESULTS: The results showed that culture media composed of milk whey without filtration supplied L. lactis its adaptation needs better than filtrated milk whey. Nisin titers, in milk whey without filtration (pH adjusted), was 11120.13 mg L^?1 in the second transfer, and up to 1628‐fold higher than the filtrated milk whey, 6.83 mg.L^?1 obtained in the first^t transfer. CONCLUSIONS: Biological processing of milk byproducts (milk whey) can be considered a profitable alternative, generating high‐value bioproducts and contributing to decreasing river disposals by dairy industries. Copyright © 2008 Society of Chemical Industry     

(S)-Reutericyclin: Susceptibility Testing and In Vivo Effect on Murine Fecal Microbiome and Volatile Organic Compounds

We aimed to assess the in vitro antimicrobial activity and the in vivo effect on the murine fecal microbiome and volatile organic compound (VOC) profile of (S)-reutericyclin. The antimicrobial activity of (S)-reutericyclin was tested against Clostridium difficile, Listeria monocytogenes, Escherichia coli, Enterococcus faecium, Staphylococcus aureus, Staphylococcus (S.) epidermidis, Streptococcus agalactiae, Pseudomonas aeruginosa and Propionibacterium acnes. Reutericyclin or water were gavage fed to male BALBc mice for 7 weeks. Thereafter stool samples underwent 16S based microbiome analysis and VOC analysis by gas chromatography mass spectrometry (GC-MS). (S)-reutericyclin inhibited growth of S. epidermidis only. Oral (S)-reutericyclin treatment caused a trend towards reduced alpha diversity. Beta diversity was significantly influenced by reutericyclin. Linear discriminant analysis Effect Size (LEfSe) analysis showed an increase of Streptococcus and Muribaculum as well as a decrease of butyrate producing Ruminoclostridium, Roseburia and Eubacterium in the reutericyclin group. VOC analysis revealed significant increases of pentane and heptane and decreases of 2,3-butanedione and 2-heptanone in reutericyclin animals. The antimicrobial activity of (S)-reutericyclin differs from reports of (R)-reutericyclin with inhibitory effects on a multitude of Gram-positive bacteria reported in the literature. In vivo (S)-reutericyclin treatment led to a microbiome shift towards dysbiosis and distinct alterations of the fecal VOC profile.     

Novel bacteriocinogenic Enterococcus hirae and Pediococcus pentosaceus strains with antilisterial activity isolated from Brazilian artisanal cheese

We isolated and characterized bacteriocin producers Enterococcus hirae ST57ACC and Pediococcus pentosaceus ST65ACC from raw milk artisanal cheeses. Their bacteriocins were tolerant to temperatures from 4°C to 100°C and under sterilization conditions (121°C for 15 min). Additionally, the tested bacteriocins remained active after being exposed to pH 2.0 to 10.0 for 2 h. The activity of the bacteriocins was affected by proteolytic enzymes but remained stable after treatment with EDTA, sodium dodecyl sulfate, NaCl, skim milk, and Tween 80. Cell-free supernatants were capable of inhibiting Listeria innocua and several strains of Listeria monocytogenes obtained from different sources and belonging to different serotypes. When L. monocytogenes 211 and L. monocytogenes 422 were treated with bacteriocins, growth was completely inhibited over 12 h. Cocultures of bacteriocinogenic strains and L. monocytogenes 422 in skim milk showed that E. hirae ST57ACC could control the growth of the pathogen in the matrix after 48 h. None of the selected isolates presented positive results on a screening panel for 25 bacteriocin-related genes, however, indicating that both strains might express novel bacteriocins.     

Different combinations of salts affect the growth and bacteriocin production by Lactobacillus salivarius CRL 1328

BACKGROUND: The culture medium for optimal growth of vaginal Lactobacillus salivarius CRL 1328 is different from that for optimal bacteriocin production. To simultaneously obtain high amount of biomass and bacteriocin of this microorganism, the effects of different basal culture media and salts on both responses were evaluated. The study was performed by using a complete factorial experimental design 2⁶, with central points. Sixty‐four different growth media, which resulted from the combinations of two basal culture media and two concentrations of five salts (ammonium citrate, sodium acetate, MgSO₄, MnSO₄, and K₂HPO₄) were assayed. RESULTS: Only the addition of MnSO₄ to each culture medium significantly stimulated the growth of L. salivarius. The presence of sodium acetate or MgSO₄ stimulated the bacteriocin production, while MnSO₄ and K₂HPO₄ exerted an inhibitory effect. However, the simultaneous addition of MnSO₄ and sodium acetate to both basal culture media allowed high bacteriocin levels to be reached, attenuating the inhibitory effect of Mn²⁺. CONCLUSIONS: The application of a complete experimental design contributed to simultaneous optimization of the biomass and bacteriocin production of L. salivarius CRL 1328. The results obtained are potentially applicable to the technological production of probiotic bacteria and antagonistic substance to be included in a probiotic pharmaceutical product. Copyright © 2009 Society of Chemical Industry     

双水相系统萃取细菌素发酵

以生长速率为标准,筛选出适合菌株乳酸乳球菌乳酸亚种ATCC 11454生长的成相聚合物聚乙二醇(PEG)20000、成相盐MgSO4.以细菌素浓度为筛选标准,优选出适合菌体生长及细菌素积累的双水相系统:PEG 20000 11%-MgSO4 3.5%.菌株细胞在该系统中完全分配在下相,虽然细胞的生长只有单相对照培养基的60%,但产物产量可提高33%.双水相系统系线和相体积比的改变均影响到菌体的生长及目的产物的积累.     

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.     

具有抑菌作用乳酸菌的筛选及其抑菌物质特性的研究

从内蒙古地区传统谷物发酵食品——酸粥中分离并鉴定的13株乳酸杆菌中筛选具有抑菌活性的菌株,并对其抑菌能力及抑菌物质的特性进行研究。结果表明,菌株NSZL-2(鼠李糖乳杆菌L.rhamnose)和NSZL-12(植物乳杆菌L.plantarum)对指示菌具有较高的抑菌活性,且抑菌能力主要来自菌体的发酵代谢产物。排除酸和过氧化氢的干扰后代谢产物仍具有抑菌活性,且抑菌物质对热具有很高的稳定性。代谢产物经蛋白酶K、胃蛋白酶、胰蛋白酶、木瓜蛋白酶、过氧化氢酶、溶菌酶处理后,抑菌能力均有不同程度的降低,说明抑菌物质对蛋白酶敏感,故判定抑菌物质中有蛋白质类细菌素。