一株强抑菌活性植物乳杆菌的分离及益生性能研究

以健康黔北黑猪肠道黏膜内容物为原料,采用厌氧罐培养法分离出肠道细菌,结合抑菌试验筛选出其中具有强抑菌活性菌种进行分子生物学鉴定,并对其耐受性及抑菌物质的稳定性进行研究。结果表明,该菌株被鉴定为植物乳杆菌（Lactobacillus plantarum）,对金黄色葡萄球菌ATCC6538、大肠埃希氏菌ATCC25922和鼠伤寒沙门氏菌ATCC14028等指示菌具有强抑制作用,其发酵上清液对3种指示菌的最大抑菌圈直径分别约为44.7 mm、23.1 mm和31.8 mm;菌株发酵上清液抑菌物质耐高温,120 ℃处理30 min抑菌活性无显著变化（P＞0.05）;胰蛋白酶处理后,上清液的抑菌圈直径显著减小（P＜0.05）,而过氧化氢酶和胃蛋白酶处理后抑菌圈直径无显著变化（P＞0.05）,表明主要抑菌物质为有机酸和活性肽;菌株经pH 2、0.3%猪胆盐或人工胃肠液处理6 h后存活率分别为0.06%、0.61%、40.80%和25.65%,显示出良好的益生性能。     

Characterization of a reuterin-producing Lactobacillus coryniformis strain isolated from a goat's milk cheese

Lactobacillus coryniformis CECT 5711, a strain isolated from a goat's milk cheese, displayed a broad-spectrum antimicrobial activity; as a consequence, its ability to produce the antagonistic compounds associated to lactic acid bacteria, including bacteriocins, hydrogen peroxide, lactic acid, acetic acid, and reuterin (3-hydroxypropionaldehyde, 3-HPA) was investigated. Production of bacteriocins or hydrogen peroxide by this strain could not be detected. However, in addition to lactic acid and acetic acid, it produced reuterin and cobalamin, a cofactor required for conversion of glycerol to 3-HPA through a glycerol dehydratase. The gene encoding a glycerol dehydratase subunit was detected by PCR and the corresponding amplicon was sequenced. This strain showed a high survival after exposition to conditions simulating those existing in the gastrointestinal tract as well as a notable ability to adhere to intestinal cells, which suggests that its reuterin-producing ability may be used for the host benefit. In addition, the strain showed a strong beta-galactosidase activity. Production of biogenic amines and degradation of mucin could not be detected.     

Isolation and characterisation of six novel mycobacteriophages and investigation of their antimicrobial potential in milk

Using Mycobacterium smegmatis as a rapidly growing host, six mycobacteriophages were isolated from a variety of environmental sources and characterised by restriction endonuclease digestion. All six phages had long, non-contractile tails and isometric heads characteristic of Siphoviridae and were relatively heat-stable up to 72 °C and generally retained infectivity at pH4–pH10 for 60 min. Each individual phage, when tested against M. smegmatis in both growth media and reconstituted milk, typically reduced host numbers by up to 7 logs. When all 6 phages were combined as a cocktail in milk, a strong bactericidal effect on M. smegmatis was observed over 96 h, evidenced by a 9-log reduction in host cell numbers. These results achieved against M. smegmatis, indicate that mycophages may have a potential application as anti-mycobacterial agents when used against more virulent pathogens such as Mycobacterium avium subsp. paratuberculosis, which has been detected in both raw and pasteurised milk.     

一株植物乳杆菌的分离鉴定及应用

从湖南自然发酵的酸豇豆中分离出一株疑似乳酸菌菌株L4,提取其16S rDNA,并经测序、同源性分析和系统发育树构建等,对其属种进行鉴定。结果表明,菌株L4为植物乳杆菌（Lactobacillus plantarum）。通过耐盐、耐酸和降解亚硝酸盐实验发现,菌株L4可在10%食盐含量条件下生长,耐受pH 2.5,对亚硝酸盐的降解率高达99%。接种菌株L4发酵萝卜干,以自然发酵萝卜干为对照,对发酵萝卜干进行感官评定初步探究该菌株发酵萝卜干的特性。结果表明,菌株L4可以缩短萝卜干发酵周期,提高发酵萝卜干感官评分。     

Anti-obesity activity of Lactobacillus fermented soy milk products

The anti-obesity activity of Lactobacillus paracasei subsp. paracasei NTU 101 and Lactobacillus plantarum NTU 102 and their soy milk fermented products (SM101 and SM102) were investigated. Results indicated that the inhibition of 3T3-L1 differentiation and the accumulation of free fatty acids markedly increased in rats treated with SM101 and SM102. Moreover, the up-regulation and down-regulation of lipolysis and heparin-releasable lipoprotein lipase, respectively, were observed in the 3T3-L1 adipocytes of the SM101 and SM101 groups, and these effects of SM101 and SM102 were greater than unfermented soy milk (USM). We also found that SM101 and SM102 both improved obesity in Wistar rats fed with a high-fat diet (HFD) and that this improvement was stronger than that observed for USM. The level of serum leptin in HFD-induced rats was significantly elevated by the 5-week administration of SM101 and SM102 (10⁶–10¹⁰ CFU/mL per rat per day); however, this activity was not promoted by USM. The anti-obesity activity of SM101 and SM102 may result from the increased daidzein and genistein levels that were observed during the fermentation by L. paracasei subsp. paracasei NTU 101 and L. plantarum NTU 102.     

泡菜中植物乳杆菌的筛选及益生活性研究

从民间自制泡菜中筛选乳酸菌。根据菌落形态,革兰氏染色和溶钙透明圈,分离得到17株乳酸菌菌株,用16S r DNA测序鉴定分离菌株,并测试了其耐酸和耐胆盐能力。结果表明,筛选菌株中WHLP-01、WHLP-02、WHLP-03和WHPE-02四株菌株可以在p H=2.0的环境下生存2 h;WHLP-01与WHLP-02两株菌株可以在0.3%胆盐浓度下存活;其中菌株WHLP-01属植物乳杆菌,其对常见的8株致病菌均有明显的抑制效果和降胆固醇能力,可以作为微生态制剂的候选菌株深入研究。      

一株高效降胆固醇嗜酸乳杆菌在发酵乳中的应用

采用高效降胆固醇嗜酸乳杆菌菌株2-2和嗜热链球菌调制发酵剂B,研究了发酵剂B在发酵乳中的应用。通过与传统保加利亚乳杆菌和嗜热链球菌调制的发酵剂A的对比,结果表明,发酵剂B有较强的胆固醇去除效力和弱的后酸化能力。4℃条件下存放15d后发酵乳的酸度为103.6°T,发酵乳中乳酸菌活菌数为2.6×107cfu/mL,高于标准的最低限制(≥106cfu/mL)。研究表明,在降胆固醇和抑制后酸化意义上,发酵剂B能够取代发酵剂A。     

瑞士乳杆菌发酵乳中生物活性小肽的免疫功能特性研究

利用MTT、中性红吞噬、一氧化氮诱生分泌等方法从已获得的瑞士乳杆菌发酵乳来源小肽中筛选出对小鼠腹腔巨噬细胞具有免疫活性的小肽GLPN,并通过ELISA方法检测TNF-Ω,IL-1β,IL-6的细胞因子水平,以对具有的免疫促进作用效果进行评估.结果表明,GLPN在0.1 g/L质量浓度下所有指标上都能表现出显著的促进作用,而在LPS作用下,GLPN质量浓度达到0.5 g/L也未引起炎症因子的过量表达.     

Coculture-inducible bacteriocin activity of Lactobacillus plantarum strain J23 isolated from grape must

Detection and characterization of bacteriocin production by Lactobacillus plantarum strain J23, recovered from a grape must sample in Spain, have been carried out. Bacteriocin activity was degraded by proteolytic enzymes (trypsin, alfa-chymotrypsin, papaine, protease, proteinase K and acid proteases), and it was stable at high temperatures (121 degrees C, 20min), in a wide range of pH (1-12), and after treatment with organic solvents. L. plantarum J23 showed antimicrobial activity against Oenococcus oeni, and a range of Lactobacillus and Pediococcus species. Bacteriocin production was detected in liquid media only when J23 was cocultivated with some inducing bacteria, and induction took place when intact cells or 55 degrees C heated cells of the inducer were cocultivated with J23, but not with their autoclaved cells. Bacteriocin activity of J23 was not induced by high initial J23 inocula, and it was detected in cocultures during the exponential phase. The presence of ethanol or acidic pH in the media reduced bacteriocin production in the cocultures of J23 with the inducing bacteria. The presence of plantaricin-related plnEF and plnJ genes was detected by PCR and sequencing. Nevertheless, negative results were obtained for plnA, plnK, plNC8, plS and plW genes.     

一株曲拉源短乳杆菌的分离及其体外抑菌特性分析

摘要：目的 天然牦牛乳曲拉中蕴含着大量的乳酸菌，为丰富食品发酵的有益菌种资源，本试验分离筛选具有抑菌活性的乳酸菌。方法 以西藏牦牛乳曲拉为分离基质，MRS(De Man Rogosa Sharpe)加上1%CaCO3为选择鉴别培养基，运用形态学观察、生理生化试验、16S r RNA基因序列、药敏试验对分离菌株进行菌种鉴定，通过抑菌试验探究其抑菌活性。结果 分离菌株NWMCC0322被鉴定为短乳杆菌，其对青霉素类、四环素类、大环内酯类、酰胺醇类、喹诺酮类、磺胺类、头孢类药物敏感，对多烯类药物中敏。NWMCC0322对指示菌金黄色葡萄球菌的抑菌效果最好，其抑菌圈直径达12.0±0.6 mm，其次为甲型副伤寒沙门氏菌11.3±0.3 mm，大肠杆菌11.0±0.5 mm。结论 该短乳杆菌对食品污染常见菌株具有抑菌效果，在食品发酵中有较好的应用价值。     

农家酸菜中优质乳酸菌的筛选分离及其产酸性能优化

采用平板画线和平板涂布方法从3种浙江西北山区民间酸菜样品中分离出6株产酸菌株,并从中优选出4株产酸能力强的菌株,经形态学及生化特性初步鉴定,4株均为乳酸杆菌.对L-3菌株的发酵产酸能力进行优化,结果表明,培养温度38℃,葡萄糖添加量14g/L,蛋白胨添加量10g／L具有最好的发酵产香、产酸性能.     

Isolation of exopolysaccharide producing Lactobacillus strains from sorghum distillery residues pickled cabbage and their antioxidant properties

Twelve Lactobacillus strains isolated from sorghum distillery residue (SDR) pickled cabbage were studied for their in vitro scavenging activity against hydroxyl radicals and DPPH free radicals, inhibition of lipid peroxidation, and their resistance to hydrogen peroxide in cell lysate or intact cells. Lactobacillus brevis D7, at a dose of 10¹⁰ CFU/mL, showed the highest hydroxyl radical and DPPH scavenging activities, as well as total antioxidative activity, with inhibition rates of 51.2, 44.9, and 65.2%, respectively in intact cells. On the other hand, the L. brevis D7 strain was the most resistant against hydrogen peroxide. L. brevis D7 isolated from SDR pickled cabbage should be considered as a potential antioxidant to be used in functional foods. Correlation analysis showed that DPPH free radical scavenging activity is a potential screening indicator for high exopolysaccharide (EPS) producing Lactobacillus species.     

The potential effect of FOS and inulin upon probiotic bacterium performance in curdled milk matrices

Inulin and fructooligosaccharides were studied for their prebiotic effect upon growth/survival of probiotic bacteria and technological potential in probiotic food processing, via characterization of glycolysis, proteolysis and lipolysis in curdled milk matrices; the ultimate goal is the manufacture of synbiotic cheeses. Prebiotic compounds did not significantly affect growth/viability of all strains studied, except Lactobacillus acidophilus La-5. Proteolysis indices revealed considerable casein degradation in probiotic and synbiotic matrices inoculated with Bifidobacterium lactis B94 and Lactobacillus casei-01; lower values were achieved in those inoculated with L. acidophilus La-5, yet a synbiotic effect was apparent in NPN values. Lipolysis was not extensive over storage, irrespective of matrix type; however, interesting differences in terms of the qualitative free fatty acids profile were observed. CLA isomers, and α-linolenic and γ−linolenic acids were detected upon 15 d of ripening of all inoculated matrices. Principal component analysis was able to discriminate the various matrices according to degree of maturation, throughout the ripening period. Microbiological and biochemical parameters unfolded a very good technological potential, especially of B. lactis B94 and L. casei-01, to produce novel types of functional dairy matrices - although extrapolation to actual cheeses should still be done with care, because e.g. syneresis was not considered.     

罗伊氏乳杆菌发酵液中具有降胆固醇能力蛋白的分离、纯化和鉴定

罗伊氏乳杆菌DSM122460无细胞上清发酵液（cell-free supernatant,CFS）具有较高的胆固醇移除能力,经初步鉴定有效成分为蛋白质。通过超滤浓缩、硫酸铵分级沉淀、DEAE Sepharose F.F阴离子交换层析进一步对该活性成分进行分离纯化。纯化后,其比活力达到61.6 U/mg,纯化倍数为32.4。经SDS-PAGE检测,样品达到电泳级纯,分子量约为60 ku。通过质谱鉴定初步认定该活性蛋白为一假设蛋白,其功能尚待进一步研究。      

酸马奶酒中乳酸菌的分离及发酵性能研究

以酸马奶酒为原料,筛选到1株优良的乳酸菌MJ3,并对该乳酸菌的形态特征、生理生化特征、培养条件、发酵性能进行了研究。结果表明,该菌为G＋短杆菌,最适生长pH为7.0,最适生长温度为35℃,最适的分离培养基是MRS培养基;该菌发酵8h时,酸度达到最大,为37.6莓T,后酸化能力弱。     

Transit of radical scavenging activity of milk products prepared by Maillard reaction and Lactobacillus casei strain Shirota fermentation through the hamster intestine

Oxidative stress in the colon is associated with the incidence of colon cancer. In situ, the suppression of oxidative stress in the colon would be an effective form of prevention of the cancer. In this study we investigated the transit of the radical scavenging activity of milk products through the hamster intestinal tract. Two types of skim milk products were prepared by Maillard reaction and then lactic acid fermentation. Heat treatment enhanced the radical scavenging activity for 2,2-diphenyl-1-picrylhydrazyl radical of skim milk. The activity was further increased by fermentation with Lactobacillus casei strain Shirota. Normal hamsters were fed these milk products for 14 d. For potential radical scavenging activity per unit dry weight of feces and cecal content, the groups ranked in the order of fermented product-fed hamsters > heated product-fed hamsters > control hamsters, reflecting the order of the potential of the corresponding diets. Approximately 12% of the activity of the heated and the fermented product diets passed through the gastrointestinal tract. These results suggest that some of the radical scavenging activity generated by food processing reached the colon in nonabsorbable products.