具有降糖作用的鼠李糖乳杆菌生物学特性研究

本研究旨在评价一株具有降糖作用的鼠李糖乳杆菌的生物学特性。结果表明,该菌株生长较快,2 h进入生长对数期,14 h后进入稳定期,最适生长p H为5⁷,最适生长温度为37℃,最适接种量为1%²%（V/V）。此外,该菌株在强酸条件下可以生长,对人工胃肠液都具有很好的耐受性,耐受胆盐的延迟时间为0.86 h,在Na Cl浓度为9%（W/V）的培养条件下仍可以存活。更为重要的是,这株菌对Caco-2细胞具有较强的粘附能力。综上所述:该鼠李糖乳杆菌菌株可作为一株具有潜在降糖作用的发酵菌株,用以开发具有降糖作用的新型发酵产品。      

鼠李糖乳杆菌LT22代谢产物抑菌实验研究

将鼠李糖乳杆菌LT22接种于发酵培养基,37℃培养48 h,离心、过滤除菌,滤液即为鼠李糖乳杆菌代谢产物(LRM).用金黄色葡萄球菌等6种菌作为指示菌,采用牛津杯双层平板定量扩散法对LRM进行体外抑菌试验.结果表明,LRM对6种指示菌有较强的抑菌活性,最大抑菌圈直径可达22.3mm,而与LRM相同pH值的乳酸、乙酸和盐酸溶液无抑菌作用;该抑菌活性物质耐热性强,不受过氧化氢酶、蛋白酶、木瓜蛋白酶、胰蛋白酶及pH值2～4变化的影响,但pH值变化到5～10时,抑菌活性却完全丧失.     

鼠李糖乳杆菌发酵液中水溶蛋白的提取及其抗氧化活性研究

采用硫酸铵沉淀法获得鼠李糖乳杆菌B10发酵液中水溶蛋白粗提物,并进行葡聚糖凝胶SephadexG-100柱纯化,分离得到不同的蛋白组分峰Ⅰ、峰Ⅱ和峰Ⅲ,分别对各组分DPPH自由基清除能力、清除超氧自由基(O2-·)和羟自由基(·OH)的能力及超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性进行分析。结果表明,纯化前后各样品均对DPPH·有清除作用,其中纯化后的峰Ⅰ对DPPH·清除能力最强,峰Ⅱ、峰Ⅲ次之,均高于水溶蛋白粗提物;纯化后的峰Ⅰ对O2-·和·OH的清除能力最强,均高于5mmol/L的Vc溶液和水溶蛋白粗提物对O2-·和·OH的清除作用,峰Ⅱ和峰Ⅲ均未检测到对O-·和·OH的清除能力。     

干酪乳杆菌纯种发酵大豆乳培养基的优化调配

从益生菌乳制品中自行分离选育出的生长繁殖力强、发酵活力高的干酪乳杆菌(05-20)为试验菌株,研究了牛乳和蔗糖在大豆乳中的添加量对干酪乳杆菌在大豆乳中发酵的凝乳时间、凝乳时活菌数、pH值、滴定酸度及产品感官风味的影响,通过方差分析确定了发酵培养基中牛乳和蔗糖的最适添加量分别为20%和7%,完成了以大豆乳作为干酪乳杆菌最佳载体的初步探索,为进一步研制开发益生菌发酵大豆乳制品奠定了基础.     

比较基因组学揭示鼠李糖乳杆菌Probio-M9在种内的基因组差异

鼠李糖乳杆菌是人和动物肠道的共生菌,在食品、医药和动物养殖等多个领域均有应用。本文以鼠李糖乳杆菌Probio-M9全基因组为对象,结合NCBI公开的214株鼠李糖乳杆菌基因组序列进行比较基因组学分析,解析不同鼠李糖乳杆菌基因组差异。结果发现,215株鼠李糖乳杆菌共识别到16 915个泛基因,247个核心基因;后续通过247个核心基因构建系统发育树,分离源和分离地不存在明显聚类趋势。鼠李糖乳杆菌Probio-M9处在分支最大的分支Ⅱ中,该分支各菌株之间的遗传关系近,差异很小,区分难度大;对分支Ⅱ中98株鼠李糖乳杆菌进行RAST注释分析,鼠李糖乳杆菌虽在功能方面整体存在高度相似性,但其中部分菌株与鼠李糖乳杆菌Probio-M9存在一定差异,相较于其它鼠李糖乳杆菌,鼠李糖乳杆菌Probio-M9具有更强的自身代谢、转录、运输等方面的调控能力,并且含有与益生功能相关的基因,如谷胱甘肽(gshAB)、胞外多糖(rmlA~rmlD、epsH)及提高宿主代谢能力(tagE)相关基因。通过比较基因组学揭示鼠李糖乳杆菌Probio-M9在种内的基因组差异,为鼠李糖乳杆菌Probio-M9的开发及应用奠定了基础。     

The involvement of ATPase activity in the acid tolerance of Lactobacillus rhamnosus strain GG

This study examined the survival and acid stress response of nine lactic acid bacteria (LAB) including Lactobacillus rhamnosus strain GG in commercially available yoghurt products (low pH environment) during the long‐term storage. Under the storage conditions investigated (52 weeks period at 5 °C), there was a loss of the viability of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium sp. within 35 weeks. However, L. rhamnosus strain GG, one of starter strains in a commercially available yoghurt product, exhibited excellent survival throughout the whole storage period. Our results indicate that the viability of L. rhamnosus strain GG was increased by modulating the stress‐related factors as well as the activity of ATPase with exposure to the low pH conditions.     

Nonfermented ice cream as a carrier for Lactobacillus acidophilus and Lactobacillus rhamnosus

Efficiency of a nonfermented ice cream for delivering Lactobacillus acidophilus and Lactobacillus rhamnosus to consumers was evaluated. Both of the microorganisms survived at the populations of greater than 10⁷ CFU g^?1 during 12 weeks of storage at ?19 °C. Addition of the microorganisms had no significant effect on the overrun, viscosity, firmness and melting behaviour; it changed the acidity, pH and sensory properties of the finished product. Resistance to acid and sensitivity to bile of both bacteria were tested separately on fresh harvested cells before inoculation to ice cream and then on the frozen‐thawed cells after 12 weeks of cold storage in ice cream. Ice cream processing followed by cold storage reduced acid resistance of both bacteria at pH 2.5. Resistance to bile in L. rhamnosus was not affected in frozen‐thawed ice cream when compared to fresh cell, whereas resistance to bile in L. acidophilus appeared to be more susceptible to the process and cold storage.     

Dry sausage fermented by Lactobacillus rhamnosus strains

The ability of three probiotic Lactobacillus rhamnosus strains GG, E-97800 and LC-705 and one commercial Pediococcus pentosaceus starter strain (control) to produce dry sausage was studied. During the fermentation process the numbers of inoculated lactic acid bacteria increased from approx. 7 log10 to 8-9 log10 cfu/g and the pH values decreased from 5.6 to 4.9-5.0. The sensory test indicated that the dry sausages fermented by L. rhamnosus LC-705 were inferior to the control sausages. The presence of inoculated experimental strains as predominant organisms in the dry sausages was recognised on the basis of their genetic fingerprints by ribotyping. The concentrations of biogenic amines remained low during the ripening process. These results indicated that the studied Lactobacillus rhamnosus strains, especially strains GG and E-97800, are suitable for use as probiotic starter cultures in fermenting dry sausage.     

产共轭亚油酸鼠李糖乳杆菌的筛选与鉴定

从东北传统酸菜汁中筛选出具有产共轭亚油酸(CLA)能力的乳酸菌菌株15株,采用紫外分光度计法测定发酵液中共轭亚油酸产量,其中菌株A53.2的产量最高,在MRS培养基中添加0.4mg/mL亚油酸(LA),37℃发酵24h,CLA产量达14.91μg/mL。通过对菌体的形态特征和生化特性分析,并结合16SrDNA分子生物学以及系统发育分析鉴定菌株A53.2为鼠李糖乳杆菌。     

微生态补钙制剂的生物利用研究

补钙制剂的生物利用率是评价补钙效果的重要指标。以KM种健康小鼠为模型,饲喂不同钙源及钙含量的饲料,测定饲料消耗量、粪钙、尿钙、骨钙和血钙,计算钙的表观吸收率及存留率,并与对应剂量的碳酸钙组对比,评价微生态补钙制剂的生物利用情况。结果表明：微生态补钙制剂低剂量组和标准剂量组的骨钙含量与相同水平的碳酸钙对照组比较差异显著(P＜0.05),钙的表观吸收率明显高于对应碳酸钙剂量组(P＜0.01);微生态补钙制剂组与碳酸钙对照组相比,血钙水平差异不显著(P＞0.05),但均高于碳酸钙对照组。表明微生态补钙制剂生物利用率高,是一种理想的补钙制剂。     

鼠李糖乳杆菌降解亚硝酸盐影响因素研究

该试验研究鼠李糖乳杆菌降解亚硝酸盐的影响因素.研究发现,鼠李糖乳杆菌降解亚硝酸盐的最适pH值是5.0～6.0,最适温度是40℃;培养基中碳源、氮源、酵母膏、金属离子成分分别为2%葡萄糖、0.5%大豆蛋白胨、0.4%酵母膏;添加Fe2+、Fe3+、Cu2+的亚硝酸盐降解效果明显;最适接种量3%;最适培养方式为动态培养.     

鼠李糖乳杆菌发酵及冻干保护的研究

对鼠李糖乳杆菌发酵性能和冷冻干燥存活率与保护剂、发酵时间和温度的关系进行了研究。利用液体发酵、紫外分光测量法、微量活菌计数法进行培养、测量。结果表明，鼠李糖乳杆菌发酵周期为12－16h；乳清粉和双歧因子对鼠李糖乳杆菌发酵产酸性能和细菌总数有定的影响。正交实验结果表明，10％乳清粉，15％脱脂奶粉，16h发酵时间的组合，冷冻干燥后存活的影响无显著性差异。结论：乳清粉和双歧因子在发酵后期会对鼠李糖乳杆菌的生长和发酵有促进作用可能是含糖量较高的原因。在冻干时，蛋白质（特别是酪蛋白）对细菌有保护作用。     

鼠李糖乳杆菌耐酸及胆盐能力研究

本实验对来源于婴儿粪便鼠李糖乳杆菌进行耐酸、耐胆盐能力表征及模拟胃液和模拟肠液实验。结果获得鼠李糖乳杆菌菌株B在pH1.8条件下,维持2h,活菌数达到107CFU/ml以上;在pH值为2.2以上的培养基中生长良好,而且随着时间的延长菌株的数量增加。在胆盐浓度在0.5%以下的MRS-broth培养基中鼠李糖乳杆菌B菌株维持4h,活菌数均达到108CFU/ml以上,而且随着时间的延长活菌数均有所增加。模拟胃液和模拟肠液实验结果表明该菌株能够有效通过胃环境,并能在肠道中繁殖。结果可见,鼠李糖乳杆菌B具有较强的酸的胆盐的耐受能力,符合微生态制剂和乳酸菌发酵功能食品菌种的要求。     

Cellular injuries and storage stability of spray-dried Lactobacillus rhamnosus GG

The objective of this work was to evaluate the applicability of spray drying in the production of skim milk-based preparations containing probiotic bacteria Lactobacillus rhamnosus GG (ATCC 53103). Furthermore, oligofructose-based or polydextrose-based prebiotic substances were also included in the carrier matrix to assess their protection capacity. When reconstituted, skim milk was used as a spray drying carrier; a microbial survival rate of 60% was achieved at an outlet temperature of 80 °C. Partial substitution of the solids content of the reconstituted skim milk by prebiotic substances also resulted in a high level of survival. However, the storage stability of the dried powder was reduced as the amount of skim milk solids in the carrier was lowered. The spray drying media were further calorimetrically characterized in terms of their glass transition temperatures so as to evaluate the contribution of glassy state on the maintenance of bacterial survival during storage. Although all evaluated carriers were in the glassy state, differences were observed in their capacity to confer protection on the probiotic bacteria. Flow cytometric assessment in combination with functional dyes was applied as a diagnostic tool to evaluate the type of cellular injuries which occurred upon spray-drying. Cell death was caused mainly by damage to cell membranes and the degree of membrane disintegration increased progressively as the outlet temperatures increased.     

酸胁迫和冷胁迫对鼠李糖乳杆菌的交叉保护作用

选用鼠李糖乳杆菌(Lactobacillus rhamnosus)hsryfm 1301为出发菌株,比较15℃低温和pH 4.0酸性条件下,对氧化、酸、渗透压和高温4种常见胁迫的耐受能力,探究低温预处理和酸性预处理对L. rhamnosus hsryfm 1301的交叉保护作用。结果表明,经过低温预处理的L. rhamnosus hsryfm 1301比对照样品在7%NaCl渗透压胁迫下的存活率提高了约高20%;L. rhamnosus hsryfm 1301经过pH4的酸性预处理比对照样品在pH3的酸性胁迫下表现出高约11%的存活率;两种预处理手段对氧化胁迫和高温胁迫均无明显交叉保护作用。     

Effects of Lactobacillus rhamnosus GG addition in ice cream

A 2⁴ full factorial experimental design was applied to verify the effects of Lactobacillus rhamnosus GG (LGG) addition in retail-manufactured ice cream stored at two different freezing temperatures (−16°C and −28°C) and containing two different levels of sugar (15–22%) and fat (5–10%). In addition to microbial counts, the pH, acidity, viscosity of the mixes and functional properties of the ice creams were evaluated. Both fresh and frozen-thawed LGG cells underwent preliminary resistance tests to bile, antibiotics and acidity. The LGG strain proved to be highly resistant to most of the stress factors. When the micro-organism was added to ice cream mixes in a quantity of 10⁸ cfu/g, it did not change the overrun, firmness or melting behaviour of the finished product. Regardless of formulation, no count decay of LGG cells was observed in ice cream stored for up to 1 year.     

碳源对鼠李糖乳杆菌生长特性的影响研究

目的研究不同鼠李糖乳杆菌菌株对不同碳源的利用及生长特性。方法在600nm波长处测定12株鼠李糖乳杆菌在不同碳源条件下的吸光度,幵采用差异分析法和分类主成分分析法对其迚行分析。结果利用特定时间点的差异分析,在葡萄糖作为主要碳源时,各菌株之间的生长情况均没有显著差异(P0.05)。12株菌在乳糖作为主要碳源时可以分为4个生长型,在蔗糖作为主要碳源时可以分为5个生长型。主成分分类法在葡萄糖作为主要碳源时,能将12株菌分为7个生长型,在乳糖作为主要碳源时可分为8个生长型,在蔗糖作为主要碳源时可分为10个类型。结论不同鼠李糖乳杆菌菌株对不同碳源的利用存在差异,表明菌株在碳源利用上具有菌株特异性,同时不同菌株利用碳源的特异性会受到碳源种类的影响。