提高酸乳中嗜酸乳杆菌活性的研究

从市售合生元中分离鉴定出嗜酸乳杆菌,以脱脂乳培养基或MRS液体培养基培养菌株,探讨嗜酸乳杆菌的生长特性,研究不同pH、保藏温度以及西红柿汁含量对嗜酸乳杆菌活性的影响。结果表明:嗜酸乳杆菌在pH6.5培养基中培养6h后的活菌数最大,且在pH2.5的培养基中0～2h内保持106CFU/mL以上,即能在较低的胃酸环境保持其益生作用;嗜酸乳杆菌酸乳在4℃下保藏活菌存活期较长;在脱脂乳培养基中添加西红柿汁对嗜酸乳杆菌的生长起促进作用,且以添加15%的西红柿汁效果最佳。     

益生菌甜酒酿的研究

以糯米经甜酒曲糖化发酵后的醪液(甜酒酿)为初始培养基,通过在其中添加各种促生长因子,对益生菌嗜酸乳杆菌的最适生长培养基及培养条件进行了研究。结果表明,在12Bx糖度的甜酒酿中加入0.1%的大豆蛋白胨和1.5%的脱脂奶粉后培养嗜酸乳杆菌,在37℃下恒温培养16～18h,其活菌数可达5.8×108cfu/ml,酸度可控制在0.5%以下。     

乳酸菌增菌培养基的营养因子优化

以改良MRS发酵培养基为基础,选择玉米浆、牛肉膏、乳糖、番茄汁、眎蛋白胨等7个营养因子增菌培养乳酸菌进行优化。利用L8(27)正交实验,优化出培养基营养因子最佳组成是:玉米浆3%、牛肉膏1%、乳糖1%。研究结果表明,嗜酸乳杆菌、嗜热链球菌、保加利亚乳杆菌、嗜酸乳酸菌,在优化后的MRS培养基发酵液中,37℃培养20 h,菌落数均高于原MRS培养基发酵液的菌落数,达到109cfu/mL以上,乳酸菌发酵液得到了浓缩,大大降低了乳酸菌发酵培养基的成本,原料成本降低了约40%。     

植物原料乳酸菌发酵剂增菌培养的研究

对筛自泡菜的嗜酸乳杆菌S1的生长条件和增殖培养基进行了研究,优化确定了乳酸菌的培养条件和增殖培养基:起始pH值为5.5、培养温度为35℃、培养时间为12h,培养基主要组成为麦芽糖2%、牛肉膏1%、缓冲盐(NaAc∶CaCO3)0.5∶0.3。控制培养条件,结合优化培养基,可使培养液活菌数提高一个数量级达到5×109cfu/mL。     

嗜酸乳杆菌、双歧杆菌和干酪乳杆菌的选择性计数

介绍了干酪乳杆菌、嗜酸乳杆菌和双歧杆菌的选择性计数方法。LC培养基只能计数干酪乳杆菌,MRS-水杨素（或山梨醇）培养基可以计数嗜酸乳杆菌和干酪乳杆菌;而MRS培养基可以计数这3种益生菌,通过减法原则从嗜酸乳杆菌、干酪乳杆菌和双歧杆菌混合物中单独计数。另外,MRS-NNLP培养基也可用于选择性计数双歧杆菌。     

混合制剂中快速计数双歧杆菌鉴别培养基的研究

研制了一种混合制剂中快速计数双歧杆菌鉴别培养基。用添加X-Gal的改良MRS培养基和厌氧胶法对4种标准双歧杆菌(长双歧杆菌、短双歧杆菌、青春双歧杆菌、两歧双歧杆菌)和2种标准乳酸菌(保加利亚乳杆菌、嗜热链球菌)分别单独培养、混合培养,结果在X-Gal培养基上,双歧杆菌单独培养及与乳酸菌混合培养双歧杆菌均为白色菌落或浅蓝色菌落,而乳酸菌为蓝色菌落。结果表明,在X-Gal培养基上双歧杆菌和乳酸菌有明显的鉴别性,可用于双歧杆菌和乳酸菌混合制剂的菌落鉴别计数。     

不同菌种发酵绿茶对其茶多酚及咖啡碱含量影响的研究

分别以保加利亚乳杆菌、嗜热链球菌、嗜酸乳杆菌三种菌种为生产菌株,以脱脂乳为培养基,茶汁添加量分别为20%、30%、40%.在37℃恒温条件下经6、12、18、24、36、48h发酵后,以紫外分光光度法测定其中茶多酚以及咖啡碱的含量.当茶汁添加量为30%,以保加利亚乳杆菌发酵12～18h时,培养基中茶多酚含量的增加速率最快.当茶汁添加量为30%,以嗜酸乳杆菌发酵12～18h时,咖啡碱含量达最大值.     

双岐杆菌酸奶的研制

就双歧杆菌以脱脂牛乳为培养基的耐氧发酵条件进行了探索,试验了导致双歧杆菌耐氧增殖的几种促进剂,且获得产菌量较高的发酵培养基,并将此菌应用于发酵乳制品。经试验确定了最佳发酵工艺,使用BAT混合菌种,或者将婴儿双歧杆菌、嗜酸乳杆菌、嗜热链球菌分别单独培养后按不同比例混合培养制备工作发酵剂,然后就可在常规条件下生产出双歧杆菌、嗜酸乳杆菌的活菌个数均高达10~(7~8)/ml以上、风味与普通酸乳相似、营养保健价值远高于普通酸乳的双歧杆菌酸奶。     

嗜酸乳杆菌培养的研究

本实验以普通营养培养基为对照,通过对pH的调节,加入不同碳源、氮源,对嗜酸乳杆菌的培养进行研究,结果表明:嗜酸乳杆菌的生长在很大程度上受到培养基的pH的影响,-般在pH 3.8的时候,嗜酸乳杆菌的生长就会受到抑制;嗜酸乳杆菌的生长也受碳源和氮源比值的限制,培养基中氮源过多,会引起微生物生长过于旺盛,不利产物的积累;如若氮源不足则菌体生长过慢,而碳源供应不足时容易引起菌体衰老和自溶.最佳培养基是:葡萄糖2%、蛋白胨2%、氯化钠0.1%、硫酸锰0.001%、硫酸镁0.02%、乙酸钠0.05%、pH6.0.     

菊粉寡糖促进嗜酸乳杆菌生长的研究

对菊粉可促进嗜酸乳杆菌的生长进行了初步研究。结果表明 ,用菊粉代替培养基中部分葡萄糖 ,嗜酸乳杆菌的生长得到明显的促进 ,当MRS培养基中葡萄糖与菊粉的质量比为 1∶1时 ,嗜酸乳杆菌的生物量及酸度得到提高 ,在 3 7℃下培养 48h ,活菌的数量及酸度比对照组分别提高 1 0 10CFU/mL和 1 0°T。     

酸奶中嗜酸乳杆菌和植物乳杆菌检测方法的研究

根据嗜酸乳杆菌、植物乳杆菌、嗜热链球菌和保加利亚乳杆菌对不同碳水化合物的选择性利用，设计了适合上述4种乳酸菌的选择性计数培养基，并对其效果进行了验证。结果表明，利用MRS培养基、麦芽糖-MRS培养基和山梨醇-MRS培养基(两种培养基共同使用)能满足选择性计数的要求。以所设计的选择性培养基，研究了含有4种乳酸菌的酸奶中不同乳酸菌在产品货架期内活菌数的变化规律。     

Probiotic bacteria: selective enumeration and survival in dairy foods

A number of health benefits have been claimed for probiotic bacteria such as Lactobacillus acidophilus, Bifidobacterium spp., and Lactobacillus casei. Because of the potential health benefits, these organisms are increasingly incorporated into dairy foods. However, studies have shown low viability of probiotics in market preparations. In order to assess viability of probiotic bacteria, it is important to have a working method for selective enumeration of these probiotic bacteria. Viability of probiotic bacteria is important in order to provide health benefits. Viability of probiotic bacteria can be improved by appropriate selection of acid and bile resistant strains, use of oxygen impermeable containers, two-step fermentation, micro-encapsulation, stress adaptation, incorporation of micronutrients such as peptides and amino acids and by sonication of yogurt bacteria. This review will cover selective enumeration and survival of probiotic bacteria in dairy foods.     

Enumeration of Lactobacillus casei in the presence of L. acidophilus,bifidobacteria and lactic starter bacteria in fermented dairy products

The aim of this work was to select a set of culture media to perform the enumeration of L. casei when it appears together with L. acidophilus, bifidobacteria and bacteria of lactic acid starters, in fermented dairy products. A number of L. acidophilus, Bifidobacterium and L. casei strains (8 of each) were tested for their ability to grow in two selective/differential media (LP-MRS agar and B-MRS agar). The enumeration of these bacteria using the media mentioned above was also carried out in fermented dairy products available on the market. B-MRS agar and LP-MRS agar were able to inhibit the lactic acid bacteria from the starters. B-MRS agar could be useful for the selective colony count of L. acidophilus or L. casei, or for a differential enumeration when these bacteria appear together in a fermented dairy product. In the case of bifidobacteria, a selective colony count could be performed on LP-MRS agar, even when it appears with L. casei, because a differential cell enumeration between both organisms was possible. The three probiotic bacteria could be simultaneously enumerated from dairy products that contain them along with starter bacteria using media developed in this work.     

Selective enumeration of probiotic microorganisms in cheese

Cheese is a dairy product which has a good potential for delivery of probiotic microorganisms into the human intestine. To be considered to offer probiotic health benefits, probiotics must remain viable in food products above a threshold level (e.g., 10⁶ cfu g⁻¹) until the time of consumption. In order to ensure that a minimal number of probiotic bacteria is present in the cheese, reliable methods for enumeration are required. The choice of culture medium for selective enumeration of probiotic strains in combination with starters depends on the product matrix, the target group and the taxonomic diversity of the bacterial background flora in the product. Enumeration protocol should be designed as a function of the target microorganism(s) to be quantified in the cheese. An overview of some series of culture media for selective enumeration of commercial probiotic cultures is presented in this review.     

新疆不同地域发酵乳品中Lactobacillus多样性的研究

利用MRS,M17等5种不同培养基从12份采自新疆北部伊犁、博乐、塔城、阿勒泰地区牧民家庭传统方法制作的乳品中分离乳酸菌,并进行了生理生化表型特征鉴定。对这些乳酸菌进行16Sr RNA基因序列的测序,构建系统发育树,初步建立其属水平的进化地位,再利用乳杆菌种间特异性引物对其进行种水平的鉴定和分类。共分离164株疑似乳酸菌,大部分菌株对温度适应性较强。以杆菌为主,系统发育表明:样品中乳酸菌主要有7个属,其中Lactobacillus(78株)、Carrobacterium(3株)、Weissella(1株)、Lactococcus(22株)、Enterococcus(47株)、Streptococcus(8株)、Vagococcus(5株)。种特性扩增显示乳杆菌存在种水平的差异。主要有4个种。利用牛津杯从样品中筛选出了10株对大肠杆菌(Escherichia coli)、枯草芽孢杆菌(Bacillus subtilis)、李斯特氏菌(Listeria monocytogenes)金黄色葡萄球菌(Staphylococcus aureus)均具有明显抑制作用的乳杆菌,为乳酸菌作为生物型防腐剂应用到食品工业中奠定基础。     

Microbial community dynamics during the Scamorza Altamurana cheese natural fermentation

The growth dynamics of the natural microbial community responsible for the fermentation of Scamorza Altamurana, a typical Southern Italian cheese made using backslopping, was investigated applying a polyphasic approach combining 1) microbial enumeration with culture media, 2) randomly amplified polymorphic DNA (RAPD) fingerprinting of microbial communities, 3) sequencing of partial 16S ribosomal DNA (rDNA) genes, and 4) physiological tests. Viable cell counts on different culture media showed that the cocci community prevailed during the 18 h of curd fermentation and the 6 d of cheese ripening. RAPD fingerprinting made it possible to isolate 25 different strains identified by 16S rDNA sequencing as belonging to five species of Lactobacillus, three species of Streptococcus, one species of Weissella, and one species of Enterococcus. The physiological analyses of all lactic acid bacteria strains revealed that the isolates belonging to Streptococcus genus were the most acidifying, whereas lactobacilli were most proteolytic. Streptococcus thermophilus C48W and Lactobacillus delbrueckii subsp. bulgaricus B15Z dominated all through the fermentation process. Furthermore, they seemed to be stable in a subsequent whey sample analyzed after 7 mo. The recovery of strains endowed with interesting technological features, such as acidifying and proteolytic activities, and surviving in natural whey could allow the upscaling of cheese processing safeguarding the organoleptic characteristics of Scamorza Altamurana and could possibly improve other fermented dairy products.     

眉山泡菜中乳酸菌的分离鉴定

目的：分析眉山泡菜的基本数据和乳酸菌的菌种构成。方法：从眉山市采集泡菜12 份,测定pH值、总酸度、盐度、乳酸菌计数,用16S rRNA序列分析法鉴定菌种。结果：样品pH值的范围是3.37～3.89,总酸度的范围是0.65～0.92 g/100 g（以乳酸计）,盐度的范围是4.16%～5.28%（以NaCl计）,MRS和M17乳酸菌计数分别是1.71～6.33、1.33～5.78（lg（CFU/g））。总酸度对乳酸菌计数影响最大。共分离鉴定出16 株乳酸菌：Lactobacillus fermentum（2 株）、Lactobacillus plantarum（3 株）、Lactobacillus brevis（2 株）、Lactobacillusacidophilus（1 株）、Lactobacillus casei（1 株）、Lactobacillus pentosus（1 株）、Lactobacillus delbrueckii（1 株）、Lactococcus lactis（3 株）、Pediococcus pentosaceus（2 株）。结论：眉山泡菜中乳酸菌存在多样性,实验结果为工业发酵生产泡菜积累了菌株。     

A comprehensive approach to determine the probiotic potential of human-derived Lactobacillus for industrial use

Specific strains should only be regarded as probiotics if they fulfill certain safety, technological and functional criteria. The aim of this work was to study, from a comprehensive point of view (in vitro and in vivo tests), three Lactobacillus strains (Lactobacillus paracasei JP1, Lactobacillus rhamnosus 64 and Lactobacillus gasseri 37) isolated from feces of local newborns, determining some parameters of technological, biological and functional relevance. All strains were able to adequately grow in different economic culture media (cheese whey, buttermilk and milk), which were also suitable as cryoprotectants. As selective media, LP-MRS was more effective than B-MRS for the enumeration of all strains. The strains were resistant to different technological (frozen storage, high salt content) and biological (simulated gastrointestinal digestion after refrigerated storage in acidified milk, bile exposure) challenges. L. rhamnosus 64 and L. gasseri 37, in particular, were sensible to chloramphenicol, erythromycin, streptomycin, tetracycline and vancomycin, increased the phagocytic activity of peritoneal macrophage and induced the proliferation of IgA producing cells in small intestine when administered to mice. Even when clinical trails are still needed, both strains fulfilled the main criteria proposed by FAO/WHO to consider them as potential probiotics for the formulation of new foods.     

Molecular quantification of lactic acid bacteria in fermented milk products using real-time quantitative PCR

Real-time quantitative PCR assays were developed for the absolute quantification of lactic acid bacteria (LAB) (Streptococcus thermophilus, Lactobacillus delbrueckii, L. casei, L. paracasei, L. rhamnosus, L. acidophilus and L. johnsonii) in fermented milk products. The results of molecular quantification and classic bacterial enumeration did not differ significantly with respect to S. thermophilus and the species of the L. casei group which were detected in the six commercial fermented products tested, thus showing that DNA extraction was efficient and that genomic DNA solutions were free of PCR inhibitors. For L. delbrueckii, the results of bacterial enumeration were generally lower by a factor 10 to 100 than those of PCR quantification, suggesting a loss of viability during storage of the dairy products at 1-8 degrees C for most of the strains in this species. Real-time quantitative assays enabled identification of the species of lactic acid bacterial strains initially present in commercial fermented milk products and their accurate quantification with a detection threshold of 10(3) cells per ml of product.     

木薯渣中乳酸菌的分离鉴定及其产酸能力分析

该研究利用传统培养法从自然发酵木薯渣中分离得到9株菌,通过菌株形态、生理生化特征及16S rDNA基因序列分析方法对菌株进行鉴定。结果显示,这9株菌均为乳杆菌属（Lactobacillus sp.）,其中菌株Y5、Y13、M14为干酪乳杆菌（Lactobacillus casei）,菌株Y6、Y12、M12为植物乳杆菌（Lactobacillus plantarum）,菌株Y7为丘状乳杆菌（Lactobacillus collinoides）,菌株M9为副干酪乳杆菌（Lactobacillus paracasei）,菌株M11为类布氏乳杆菌（Lactobacillus parabuchneri）;产酸及耐酸试验结果表明这5种菌均具有较强的产酸能力及耐酸性,发酵6 h后发酵液中pH值可达4.5以下,其中菌株Y12和M14的产酸能力最强,在15 h后其pH值均在3.6以下,可用于木薯渣或其他青贮饲料发酵剂的研制。