Lactic acid bacteria isolated from raw and fermented pork products: Identification and characterization of catalase-producing Pediococcus pentosaceus

Food Science and Biotechnology - Lactic acid bacteria (LAB) from raw and fermented pork samples were screened for their inhibitory activity by an agar spot test in order to obtain a LAB strain with...     

Pediococcus pentosaceus NB-17 for probiotic use

The plant-derived Pediococcus pentosaceus NB-17 was isolated from Japanese traditional vegetable pickles. The production of cytokines from mouse spleen cells co-cultivated with heat-killed bacteria was investigated in vitro. The bacteria significantly induced secretion levels of interferon (IFN)-gamma and interleukin (IL)-12 p70, and suppressed IL-4 productions in ovalbumin (OVA) sensitized mouse spleen cells. Therefore, the bacteria could effectively stimulate immune activities and showed allergic inhibitory effects. Further study on acid tolerance was performed under simulated gastric conditions and NB-17 showed resistance to simulated gastric acidity at pH 3.0 and pH 2.5. Moreover, after oral administration of the intact cells to rats, bacterial colonies derived from feces were analyzed by random amplification polymorphic DNA (RAPD). It was confirmed that the administered strain NB-17 remained alive in feces. These results suggest the possibility to use the P. pentosaceus NB-17 as functional foods.     

Reconstitution and function of Tetragenococcus halophila chaperonin 60 tetradecamer

Tetragenococcus halophila originally isolated from soy sauce is a halophilic lactic acid bacterium which can grow under 4 M sodium chloride. T. halophila chaperonin composed of a core moiety of chaperonin 60 (cpn60) and a lid moiety of chaperonin 10 (cpn10), is thought to contribute to host halotolerant capability. In this report, we reconstituted and characterized the core complex of T. halophila chaperonin by using a recombinant T. halophila cpn60 (Tcpn60) overexpressed in Escherichia coli. The reconstitution of Tcpn60 was performed in the presence of 10 mM MgCl2, 2 mM ATP and 0.8 M (NH4)2SO4 and the resultant oligomer was purified by gel filtration chromatography. Electron microscopy of the reconstituted Tcpn60 revealed a double toroidal tetradecameric structure that is characteristic of bacterial cpn60. The T. halophila tetradecamer cpn60 exhibited an ATPase activity and a refolding activity of both chemically and thermally denatured enolases under wide range of salt concentrations. Furthermore, we demonstrated that heterologous expression of Tcpn60 allowed the normal growth of host Escherichia coli cells under salt stress conditions and this effect was further enhanced by co-expression with Tcpn10. These results suggested that Tcpn60 contributes to the halotolerance property of T. halophila cell as a tetradecamer complex, probably associated with the Tcpn10 complex.     

戊糖片球菌表面蛋白性质及抑菌作用研究

以戊糖片球菌(Pediococcus pentosaceus) F28-8为研究对象,菌株的生长情况与产酸能力分析表明,F28-8具有良好的酸适应能力,透射电镜(TEM)观察表明其存在蛋白层结构。应用5 mol/L LiCl提取菌株表面蛋白并进行SDS-PAGE和MALDI-TOF-MS分析,发现该菌株表面含有主要组分为分子量83.5 kDa的N-乙酰基胞壁酿-L-丙氨酸酰胺酶和分子量51. 1 kDa的LysM肽聚糖结合域蛋白。进一步的菌株表面性质测试、胶原蛋白黏附实验以及抑菌实验结果显示,表面蛋白的剥离显著降低了菌株F28-8的表面性质(自动聚集能力、表面疏水性和表面电荷),降幅达到19.0%～56. 1%;LiCl提取的F28-8表面蛋白能显著降低沙门氏菌(Salmonella enterica subsp. CICC 21513)和金黄色葡萄球菌(Staphylococcus aureus CICC 21600)对胶原蛋白的黏附力,降幅分别为74. 59%和81. 16%;LiCl提取的F28-8表面蛋白对沙门氏菌和金黄色葡萄球菌的抑菌率分别达到31. 5%和15. 6%。研究结果表明,戊糖片球菌F28-8表面蛋白能改变菌体表面性质,增强其抑制致病菌黏附的能力并表现出一定的抑菌能力,在抗菌剂开发领域具有一定的应用潜力。     

Potential of wine-associated lactic acid bacteria to degrade biogenic amines

Some lactic acid bacteria (LAB) isolated from fermented foods have been proven to degrade biogenic amines through the production of amine oxidase enzymes. Since little is known about this in relation to wine micro-organisms, this work examined the ability of LAB strains (n = 85) isolated from wines and other related enological sources, as well as commercial malolactic starter cultures (n = 3) and type strains (n = 2), to degrade histamine, tyramine and putrescine. The biogenic amine-degrading ability of the strains was evaluated by RP-HPLC in culture media and wine malolactic fermentation laboratory experiments. Although at different extent, 25% of the LAB isolates were able to degrade histamine, 18% tyramine and 18% putrescine, whereas none of the commercial malolactic starter cultures or type strains were able to degrade any of the tested amines. The greatest biogenic amine-degrading ability was exhibited by 9 strains belonging to the Lactobacillus and Pediococcus groups, and most of them were able to simultaneously degrade at least two of the three studied biogenic amines. Further experiments with one of the strains that showed high biogenic amine-degrading ability (L. casei IFI-CA 52) revealed that cell-free extracts maintained this ability in comparison to the cell suspensions at pH 4.6, indicating that amine-degrading enzymes were effectively extracted from the cells and their action was optimal in the degradation of biogenic amines. In addition, it was confirmed that wine components such as ethanol (12%) and polyphenols (75 mg/L), and wine additives such as SO₂ (30 mg/L), reduced the histamine-degrading ability of L. casei IFI-CA 52 at pH 4.6 by 80%, 85% and 11%, respectively, in cell suspensions, whereas the reduction was 91%, 67% and 50%, respectively, in cell-free extracts. In spite of this adverse influence of the wine matrix, our results proved the potential of wine-associated LAB as a promising strategy to reduce biogenic amines in wine.     

发酵剂戊糖片球菌I9应用于发酵香肠的效果分析

从300余个菌落中分离得到120株“纯菌落”菌株。通过对纯菌落生理生化特征等的分析,确定所得的其中一株纯菌落为戊糖片球菌19（Pediomccus pentcvsaceus 19）。并对其在发酵香肠中的效果进行了初步分析。     

味全乳酸菌

乳品是消费者每天都需要摄入的东西,但因消费者自身原因会在摄入后产生不适的状况．加之最近频繁爆出乳品安全问题,让消费者对乳品的选择有了一些担忧．所以纯奶制品的销售额一直不太稳定,这给纯奶企业与经销商的投资带来了不利因素。但如果不是纯奶制品的乳品．例如果奶、花生奶等,却一直没有在销售业绩上出现下滑征兆。     

The effect of Pediococcus damnosus and Pediococcus pentosaceus on the growth of pathogens in minced meat

The antibacterial effects of one strain of Pediococcus damnosus and two strains of Pediococcus pentosacaeus against Clostridium perfringens, Listeria monocytogenes, Salmonella infantis and Yersinia enterocolitica were investigated. Growth inhibition studies were conducted in juice from minced meat incubated at +6 degrees C and +15 degrees C for various periods after the inoculation with pediococci. Inhibitory effects were seen for all bacteria tested.     

乳酸菌冷冻干燥保护剂的筛选

通过单因素比较和正交试验设计,确定适合保加利亚乳杆菌和嗜热链球菌的最佳冻干保护剂配方。通过对15种冻干保护剂单因素保护效果的比较,以及对选定的4种保护剂的正交试验结果,可以确定嗜热链球菌最佳冻干保护剂配方为脱脂乳粉12%、海藻糖1%、甘油3%、谷氨酸钠1%,保加利亚乳杆菌最佳冻干保护剂配方为脱脂乳粉8%、海藻糖1%、甘油2%、谷氨酸钠1.5%。     

Tolerance to high osmolality of the lactic acid bacterium Oenococcus oeni and identification of potential osmoprotectants

Growth of the lactic acid bacterium Oenococcus oeni under hyperosmotic constraint was investigated in a chemically defined medium. The bacterium could grow on media with an elevated osmolality, preferably below 1.5 Osm kg(-)(1) H(2)O. At osmolalities comprised between 0.6 and 1.5 Osm kg(-)(1) H(2)O, the growth deficit elicited by the sugars glucose and fructose was slightly more severe than with salts (NaCl or KCl). In contrast to what was observed in other lactic acid bacteria, proline, glycine betaine and related molecules were unable to relieve inhibition of growth of O. oeni under osmotic constraint. This was correlated to the absence of sequences homologous to the genes coding for glycine betaine and/or proline transporters described in Lactococcus lactis and Lactobacillus plantarum. The amino acid aspartate proved to be osmoprotective under electrolyte and non-electrolyte stress. Examination of the role of peptides during osmoregulation showed that proline- and glutamate-containing peptides were protective under salt-induced stress, and not under sugar-induced stress. Under high salt, PepQ a cytoplasmic prolidase that specifically liberated proline from di-peptides increased activity, while PepX (X-prolyl-dipeptidyl aminopeptidase) and PepI (iminopeptidase) activities were unaffected. Our data suggest that proline- and glutamate-containing peptides may contribute to the adaptation of O. oeni to high salt through their intracellular hydrolysis and/or direct accumulation.     

泡菜用乳酸菌富集培养的研究

泡菜用直投式乳酸菌发酵剂制作中,菌体富集和冻干浓缩是关键点。主要以实验室现有的3种菌株:植物乳杆菌、肠膜明串珠菌、短乳杆菌为出发菌株,进行富集培养的研究,采用缓冲盐法和化学中和法2种培养方式。通过缓冲盐法确定了植物乳杆菌、肠膜明串珠菌、短乳杆菌的最适pH均为7.0,最适培养温度分别为30℃、30℃、35℃;化学中和法采用控制pH为7.0,培养温度分别为30℃、30℃、35℃的培养条件。通过2种方法的比较,可知化学中和法得到的活菌数约为缓冲盐法的3～5倍。     

泡菜中植物乳酸菌的筛选与鉴定

目的:对自然发酵泡菜盐水中的乳酸菌进行分离鉴定及特性研究。方法:乳酸菌分离采用倾注法使稀释样品与MRS培养基混合,然后在37℃中培养,再挑取乳酸菌在MRS培养基上进行划线分离。结果:试验成功地从泡菜盐水中分离出一种植物乳酸菌菌株,并对其进行了形态特征、培养特征和生化特征鉴定,也对其生长曲线进行了测定和分析。在生化特征鉴定中,进行了革兰氏染色、H2S检验、明胶液化试验、H2O2试验、乳酸定性试验、碳水化合物发酵试验、精氨酸水解试验、运动性试验,根据结果可初步确定为植物乳杆菌。其生长曲线也满足细菌的典型生长曲线,有明显的延滞期、对数生长期、稳定期和衰亡期。结论:其在对数生长期时,pH变化最大,说明菌株的产酸性能迅速得到了提高。在稳定期时,该菌的产酸量达到最大值。     

培养时间对乳酸菌自溶影响及菌体形态学变化

乳酸菌自溶对干酪的成熟有着及其重要的意义，但对生产发酵剂却有较大的负面影响。研究了培养时间对乳酸杆菌LB-3自瘩行为的影响．研究结果表明，在迟滞期向对数生长期的过渡时期乳酸杆菌LB-3的自溶度最大，有73％的菌体发生自溶。同时利用透射电镜对乳酸杆菌LB-3在自溶期间菌体细胞所发生的形态学变化进行了观察，在自溶过程中乳酸杆菌LB-3菌体细胞变得透明，细胞质浓缩．而且部分细胞壁被自溶酶水解掉。     

Potential of lactic acid bacteria isolated from specific natural niches in food production and preservation

Autochthonous strains of lactic acid bacteria (LAB) have been isolated from traditionally homemade cheeses collected from specific ecological localities across Serbia and Montenegro. Genetic and biochemical analysis of this LAB revealed that they produce bacteriocins, proteinases and exopolysaccharides. LAB produces a variety of antimicrobial substances with potential importance for food fermentation and preservation. Apart from the metabolic end products, some strains also secrete antimicrobial substances known as bacteriocins. Among the natural isolates of LAB from homemade cheeses, bacteriocin producers were found in both lactococci and lactobacilli. Lactococcus lactis subsp. lactis BGMN1-5 was found to produce three narrow spectrum class II heat-stable bacteriocins. In addition to bacteriocin production, BGMN1-5 synthesized a cell envelope-associated proteinase (CEP) and shows an aggregation phenotype. Another isolate, L. lactis subsp. lactis BGSM1-19 produces low molecular mass (7 kDa) bacteriocin SM19 that showed antimicrobial activity against Staphylococcus aureus, Micrococcus flavus and partially against Salmonella paratyphi. Production of bacteriocin reaches a plateau after 8 h of BGSM1-19 growth. Bacteriocin SM19 retained activity within the wide pH range from 1 to 12 and after the treatment at 100 degrees C for 15 min. Among collection of lactobacilli, the isolate Lactobacillus paracasei subsp. paracasei BGSJ2-8 produces heat-stable bacteriocin SJ (approx. 5 kDa) polypeptide. It retained activity after treatment for 1 h at 100 degrees C, and in the pH range from 2 to 11. In addition to isolates from cheeses, bacteriocin-producing human oral lactobacilli were detected. Most of them showed antimicrobial activity against streptococci, staphylococci and micrococci, but not against Candida. Isolate BGHO1 that showed the highest antimicrobial activity was determined as L. paracasei. Interestingly, Lactobacillus helveticus BGRA43, which was isolated from the human intestine showed strong activity against Clostridium sporogenes, but it was not possible to detect any bacteriocin production in this isolate by using standard procedures. Further analysis of antimicrobial activity revealed that BGRA43 has a relatively broad spectrum. Lactobacilli resistant to nisin were also detected among natural isolates. They produce bacteriocins, which have no activity against nisin producing lactococci.     

乳酸菌的生物拮抗作用

论述了乳酸菌的生物拮抗作用,包括对致病菌和乳酸菌之间的拮抗。并简要论述了产生拮抗作用的可能因素或机理,为乳酸菌进一步开发提供一种理论依据。     

具抗氧化活性乳酸菌的筛选

以体外过氧化氢耐受能力为指标,从发酵食品中筛选出2株能够耐受1mmol/L浓度过氧化氢的乳酸菌,两株细菌的耐过氧化氢能力与L. rhamnosusGG的耐过氧化氢能力相当。研究了两株乳酸菌完整细胞和无细胞提取物的DPPH自由基和羟自由基清除能力。结果表明,菌体浓度为109mL-1的L1001完整细胞和无细胞提取物的DPPH自由基清除率分别为36.2%和37.9%,同样浓度的F12完整细胞和无细胞提取物对DPPH自由基的清除率分别为24.4%和27.0%;清除羟自由基实验表明,细胞浓度为109mL-1的细胞清除羟自由基能力与同体积的浓度为1mmol/L抗坏血酸相当,L1001的羟自由基清除能力优于F12。利用API鉴定系统L1001菌株被鉴定为植物乳杆菌Lactobacillus plantarum L1001。     

干酪中非发酵剂乳酸菌

非发酵剂乳酸菌是干酪中主要的次生菌群，不属于发酵微生物，通常不能很好的在牛奶中生长，不能产酸，但对干酪的风味形成有很重要作用。本文概述了非发酵剂乳酸菌的一些特征，包括不同原料制成的干酪中非发酵剂乳酸菌的来源、生长能源的利用、自身存在的蛋白分解系统对干酪风味形成影响及作为一种提高干酪品质的附属发酵剂的应用展望。