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。     

Screening of lactic acid bacteria and bifidobacteria for potential probiotic use in Iberian dry fermented sausages

The purpose of this study was to select lactic acid bacteria and bifibobacteria strains as potential probiotic cultures during the processing of Iberian dry fermented sausages. A total of 1000 strains were isolated from Iberian dry fermented sausages (363), and human (337) and pig faeces (300) in different culture media. Around 30% of these strains, mainly isolated from Iberian dry fermented sausages in LAMVAB agar, were pre-selected for testing as potential probiotics by their ability to grow adequately at the pH values and NaCl concentrations of these meat products during the ripening process. Of the in vitro investigations used to predict the survival of a strain in conditions present in the gastro intestinal tract, exposure to pH 2.5 showed itself to be a highly discriminating factor with only 51 out of 312 pre-selected strains resisting adequately after 1.5h of exposure. All acid-resistant isolates identified as lactobacilli originated from human faeces (Lactobacillus casei and Lactobacillus fermentum) and pig faeces (Lactobacillus reuteri, Lactobacillus animalis, Lactobacillus murinus, and Lactobacillus vaginalis). Pediococcus acidilactici strains were isolated from Iberian dry fermented sausages and pig faeces, whereas the greatest number of Enterococcus strains were identified as Enterococcus faecium, with this species being isolated from Iberian dry fermented sausages, and human and pig faeces. Most of these strains are promising probiotic meat culture candidates suitable for Iberian dry fermented sausages.     

植物乳杆菌发酵梨原汁饮料的研究

乳酸菌发酵果蔬汁,因其绿色、健康、营养,成为日常生活中必不可少的食物。我国梨种植业广泛、产量高,并且梨果自身营养素全面。因此,该研究利用3种复合益生菌研制一款发酵梨浆饮料。对发酵优势菌株进行发酵剂制备并优化工艺,保护剂配比为:谷氨酸钠2%、海藻糖5%、乳糖5%、脱脂乳10%、保护剂添加量与菌泥为3∶1(mL/g),在这些条件下,得到菌粉发酵剂菌活为8.48×10^9cfu/g;在发酵梨浆应用的工艺进行优化,最佳工艺为:植物乳杆菌299、保加利亚乳杆菌717、嗜热链球菌176体积比为4∶1∶1、接种量为8%、发酵温度为37℃、发酵时间72 h。此条件下梨浆得到充分发酵,口感突出,风味酸甜;测定梨浆发酵前后有机酸的变化:酵前梨浆中含量较为丰富的有机酸为:柠檬酸、苹果酸、乳酸、草酸。其中苹果酸、柠檬酸有优势有机酸,含量分别为:36.49、38.59 mg/100 g。经3种复合乳酸菌发酵后,梨浆中有机酸含量发生变化,有机酸含量依次为:乳酸、柠檬酸、苹果酸、草酸。其中乳酸含量由1.65 mg/100 g升高为116.27 mg/100 g。     

Effects of rare sugar D-allulose on acid production and probiotic activities of dairy lactic acid bacteria

It has recently been reported that the rare sugar d-allulose has beneficial effects, including the suppression of postprandial blood glucose elevation in humans, and can be substituted for sucrose as a low-calorie food ingredient. To examine the applications of d-allulose in the dairy industry, we investigated the effects of d-allulose on the acid production of 8 strains of yogurt starter (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and 4 strains of lactococci, including potential probiotic candidates derived from dairy products. Acid production by 2 L. delbrueckii ssp. bulgaricus yogurt starter strains in milk was suppressed by d-allulose, but this phenomenon was also observed in some strains with another sugar (xylose), a sugar alcohol (sorbitol), or both. In contrast, among the dairy probiotic candidates, Lactococcus lactis H61, which has beneficial effects for human skin when drunk as part of fermented milk, was the only strain that showed suppression of acid production in the presence of d-allulose. Strain H61 did not metabolize d-allulose. We did not observe suppression of acid production by strain H61 with the addition of xylose or sorbitol, and xylose and sorbitol were not metabolized by strain H61. The acid production of strain H61 after culture in a constituted medium (tryptone–yeast extract–glucose broth) was also suppressed with the addition of d-allulose, but growth efficiency and sugar fermentation style were not altered. Probiotic activities—such as the angiotensin-converting enzyme inhibitory activity of H61-fermented milk and the superoxide dismutase activity of H61 cells grown in tryptone–yeast extract–glucose broth—were not affected by d-allulose. d-Allulose may suppress acid production in certain lactic acid bacteria without altering their probiotic activity. It may be useful for developing new probiotic dairy products from probiotic strains such as Lactococcus lactis H61.     

Immunomodulating effects of peptidic fractions issued from milk fermented with Lactobacillus helveticus

The effect of peptides released during the fermentation of milk on the humoral immune system and on fibrosarcoma growth was studied. Lactobacillus helveticus was able to release peptidic compounds during milk fermentation due to its high proteolytic activity, as was shown by the degree of proteolysis and size-exclusion HPLC elution profiles. Three fractions of these compounds were separated and fed to mice during different periods (2, 5, and 7 d). The humoral immune response was assessed by following the number of IgA-secreting cells, and the antitumor activity was monitored by studying the regression of subcutaneously implanted fibrosarcomas. Feeding during 2 and 7 d with the medium-sized fraction (Fraction II) significantly increased the IgA-producing cells in the intestines, whereas feeding with the large compound fraction (Fraction I) during 5 d and the small compound fraction (Fraction III) during all three feeding periods provided similar increases. A double dose of Fraction II showed the highest IgA-producing cell count. The increase by Fraction III was shown to be caused by the presence of L-Tryptophan. Fraction II significantly decreased the size of fibrosarcoma when previously fed during 7 d, and feeding with Fraction I during 5 d decreased significantly its size after 35 d of growth. Although the mechanisms by which lactic acid bacteria enhance the immune system are not clear, this study clearly shows that bioactive compounds released in fermented milks contribute to the immunoenhancing and antitumor properties of these products. The release of bioactive peptides by lactic acid bacteria can have important implications on the modulation of the cellular immune response.     

自然发酵辣椒中乳酸菌的分离及其发酵性能研究

本文简单介绍了从自然发酵辣椒中分离和鉴定优良乳酸菌的方法。从中选育出三株乳酸菌Lact 1和Lact 2，Lact 3，并对这三株菌株的特性进行了研究。结果表明：Lact 1、Lact 2和Lact 3这三株乳酸菌在产酸速度、耐盐性等方面都适合用来发酵辣椒。     

Assessment of potential probiotic properties of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains isolated from kefir

In this study, the metabolic activities (in terms of quantities of the produced lactic acid, hydrogen peroxide, and exopolysaccharides) of 8 strains of Lactobacillus spp., Lactococcus spp., and Pediococcus spp., were determined. Lactic acid levels produced by strains were 8.1 to 17.4 mg/L. The L. acidophilus Z1L strain produced the maximum amount (3.18 μg/mL) of hydrogen peroxide. The exopolysaccharides (EPS) production by the strains was ranged between 173 and 378 mg/L. The susceptibility of 7 different antibiotics against these strains was also tested. All strains were found to be sensitive to ampicillin. The tolerance of the strains to low pH, their resistance to bile salts of strains, and their abilities to autoaggregate and coaggregate with Escherichia coli ATCC 11229 were also evaluated. High EPS-producing strains showed significant autoaggregation and coaggregation ability with test bacteria (P < 0.01). A correlation also was determined between EPS production and acid-bile tolerance (P < 0.05). EPS production possibly affects or is involved in acid-bile tolerance and aggregation of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains and supports the potential of L. acidophilus Z1L strain as new probiotic.     

发酵蔬菜源食品用益生乳酸菌的筛选及降胆固醇能力评价

为了从发酵蔬菜中筛选出具有降解胆固醇能力的食品用益生乳酸菌,作者以融合魏斯氏菌（Weissella confuse) RW作为对照,采用定向分离法结合16S rDNA分析不同来源的发酵蔬菜样品中得到5株具有降胆固醇能力的益生乳酸菌。结果表明,5株菌对pH 2.0、质量浓度为3 g/L的胆盐以及模拟胃肠液均具有耐受性,降胆固醇能力强（54.79%~62.54%）、能够抑制病原菌且黏附于Caco-2细胞上,表现出优良的益生特性。此外,5株菌均无溶血现象,不产生物胺,对多数抗生素敏感,安全性高。通过16S rDNA鉴定5株菌分别为植物乳植杆菌（Lactiplantibacillus plantarum）JCSMC-1、植物乳杆菌（Lactobacillus plantarum）SYB和JCSMC6-2、副干酪乳酸菌（Lacticaseibacillus paracasei）SYD和XSMC-1。本研究获得的1株植物乳植杆菌、2株植物乳杆菌、2株副干酪乳酸菌均具有食品用益生菌的基本特性,为开发食品用益生菌提供了新资源。     

乳酸菌发酵辣椒泡菜工艺的研究

研究了辣椒泡菜在不同的蔗糖添加量、接种量及食盐添加量的条件下对辣椒泡菜发酵时总酸的影响,并在此基础之上,通过正交试验确定发酵辣椒泡菜最优工艺条件为蔗糖添加量为3％ (w/v),接种量为8％ (v/v),食盐添加量为4％ (w/v),在20℃～25℃发酵7d～15d.通过质构及有机酸分析,结果显示用此方法发酵的辣椒,质地优良、产酸率高.此外,辣椒中的亚硝酸盐含量远低于国家标准.因此,本发酵工艺具有应用于工业化生产的前景.     

发酵鱼制品中乳酸菌的筛选鉴定及其初步应用

自传统发酵鱼制品中分离得到98株乳酸菌,通过发酵适应性和发酵特性等试验筛选出2株优良乳酸菌Z40和Y33,其具有良好的生长特性、产酸性能和耐盐性;经16S rDNA序列分析鉴定,乳酸菌Z40和Y33均为植物乳杆菌。将乳酸菌Z40应用于鱼糜发酵,结果表明该菌可快速降低鱼糜体系的pH值,显著抑制挥发性盐基氮的形成。     

Screening and Application from Fermented of Lactic Acid Bacteria Fish Products

自传统发酵鱼制品中分离得到98株乳酸菌,通过发酵适应性和发酵特性等试验筛选出2株优良乳酸菌Z40和Y33,其具有良好的生长特性、产酸性能和耐盐性;经16SrDNA序列分析鉴定,乳酸茵Z40和Y33均为植物乳杆菌。将乳酸菌Z40应用于鱼糜发酵,结果表明该菌可快速降低鱼糜体系的pH值,显著抑制挥发性盐基氮的形成。     

发酵羊奶中乳酸菌耐热菌株的筛选

对从商业乳酸菌发酵剂分离纯化的11株保加利亚乳杆菌和11株嗜热链球菌在发酵羊奶中的耐热性进行了研究。结果表明,保加利亚乳杆菌L.b-346和L.b-124菌株、嗜热链球菌S.t-883菌株和S.t-124菌株的耐热性较好;当L.b-346菌株、L.b-124菌株与S.t-883菌株、S.t-124菌株按杆菌和球菌以不同比例配合时,L.b-346菌株与S.t-124菌株以1∶2比例配合,可获得较高的耐热性。     

发酵羊乳中乳酸菌低产粘菌株的筛选

对从商业乳酸菌发酵剂分离纯化的11株保加利亚乳杆菌和11株嗜热链球菌在发酵羊乳中的粘度进行了研究。结果表明,保加利亚乳杆菌L.b-124菌株和L.b-346菌株、嗜热链球菌S.t-222菌株和S.t-346菌株具有较低的产粘能力,凝乳时间也较短;将L.b-346菌株与S.t-346菌株按杆菌和球菌1:1比例混合后在43℃下发酵羊乳时,可获得较低的发酵粘度。     

接种植物乳杆菌和蔗糖添加量对发酵香肠品质特性的影响

研究接种植物乳杆菌6003和蔗糖添加量（0.4%和1.5%）对发酵香肠品质的影响。结果表明：接种植物乳杆菌6003,可加速发酵过程中pH值的降低;发酵香肠的硫代巴比妥酸反应物（thiobarbituric acid reactive substances,TBARs）值和总挥发性盐基氮（total volatile basic nitrogen,TVB-N）含量显著降低（P＜0.05）;发酵香肠菌落总数和乳酸菌数在发酵过程中迅速增加（P＜0.05）,在贮藏期间显著减少后趋于平稳,且菌落总数和乳酸菌数显著高于自然发酵香肠（P＜0.05）;此外,接种后的发酵香肠具有更高的硬度及总体可接受度评分,颜色无显著变化。发酵后,随着贮藏时间的延长,各组发酵香肠的pH值和可滴定酸度无显著变化,黏聚性、亮度值、红度值和水分活度显著降低,黄度值、TBARs值、TVB-N含量、硬度、咀嚼度和胶着度显著升高;不同添加量蔗糖对发酵香肠相关参数的影响较小。因此,选择蔗糖添加量0.4%和植物乳杆菌6003接种量7（lg（CFU/g））的方法生产发酵香肠较为适宜。     

传统发酵蔬菜中拮抗温和气单胞菌的乳酸菌的筛选与鉴定

目的:从传统发酵蔬菜中筛选对温和气单胞菌具有拮抗作用的乳酸菌。方法:采用双层琼脂扩散法筛选,通过生理生化反应和16S rRNA序列分析进行鉴定,研究蛋白酶、p H和温度等因素对乳酸菌无细胞上清液（CFS）抑菌活性的影响,采用扫描电镜分析乳酸菌无细胞上清液对温和气单胞菌细胞结构完整性的影响。结果:从腌渍酸黄瓜中筛选出对温和气单胞菌（10⁶CFU/m L）具有较强抑制活性的菌株LZH2-5,经鉴定为植物乳杆菌（Lactobacillus plantarum）。LZH2-5无细胞上清液经胰蛋白酶、木瓜蛋白酶、中性蛋白酶、碱性蛋白酶和胃蛋白酶处理后其抑菌活性分别丧失29.38%、23.93%、22.30%、19.47%和14.59%;经40、60、80℃处理30 min后其拮抗活性基本不变,在100℃和121℃处理30 min后活性分别下降了16.66%和25.40%,在p H3.0⁴.0范围内保持其抑菌活性,粗提物的最小抑菌浓度12.0 mg/m L。扫描电镜表明经LZH2-5 CFS处理使温和气单胞菌的细胞结构破坏并溶解。结论:从腌渍酸黄瓜中筛选的植物乳杆菌LZH2-5对温和气单胞菌具有较强抑制作用,初步分析抑菌活性物质为细菌素类,可作为水产品养殖过程中控制温和气单胞菌的拮抗制剂出发菌株。      

龙眼混合果蔬汁乳酸菌饮料的生产工艺

研究了重庆市涪陵区所产龙眼混合果蔬汁乳酸菌发酵饮料的工艺条件,以优质龙眼、胡萝卜、番茄汁为原料,用德氏乳杆菌保加利亚亚种和嗜热链球菌为菌种,进行菌种活化和驯化。确定了菌种、中间种子扩大液、发酵液及发酵饮料口感稳定性的工艺配方。结果显示,用德氏乳杆菌保加利亚亚种和嗜热链球菌按1∶1比例作为菌种;种子扩大液的最佳配方为龙眼、胡萝卜、番茄混合汁配比3∶6∶1,3%葡萄糖,4%脱脂乳,2%接种量;发酵液配方:龙眼、胡萝卜、番茄混合汁配比3.5∶5∶1.5,发酵温度37℃,发酵时间24 h,接种量4%;发酵饮料调配配方:发酵原液中添加蔗糖6%,柠檬酸0.04%,CMC-Na 0.1%,黄原胶0.01%。     

Screening and Identification of Functional Lactobacillus Specific for Vegetable Fermentation

Abstract: In this study, functional lactic acid bacteria (LAB) specific for vegetable fermentation were screened from Chinese sauerkraut brine by designing purpose media. The selected LAB strains showed excellent tolerance to high concentrations of bile salt and acids. They can also survive passage through the simulated gastric fluid and may be able to reach to the intestine. Through the performance measurements of vegetable fermentation, one of these strains, NCU116 was found to have the best fermentation ability, and further subjected to a series of analyses including identification of physiological and biochemical characteristics, 16S rDNA and pheS sequencing. Taken together, the strain NCU116 was identified as Lactobacillus plantarum subsp. plantarum. These results laid a good foundation for the fermentation research in the future.     

潜在益生乳酸菌分离和鉴定研究进展

利用益生菌提高人类健康水平是当前营养健康领域最具前景的方向之一。乳酸菌是食品中常用的益生菌,迄今为止,联合国粮食及农业组织（FAO）和世界卫生组织（WHO）推荐仅从人的胃肠道分离的乳酸菌可在食品中使用。而越来越多的研究从食品来源中分离出具有益生功能的菌株以扩大益生菌的选择范围。随着分子生物学和检测分析等技术的发展,基质辅助激光解吸电离-飞行时间质谱（MALDI-TOF MS）、全基因组测序、实时荧光定量聚合酶链式反应（RT-FQ-PCR）等技术可以鉴定到亚种及菌株水平上的益生菌。鉴于分离鉴定技术是益生菌安全规范应用的先决条件,该文综述了近年来针对潜在益生乳酸菌有效分离和鉴定技术的研究进展,以期为我国益生乳酸菌自主开发利用提供借鉴。     

新疆酸驼乳中细菌素乳酸菌的筛选及其抑菌性

通过双层琼脂扩散打孔法,从新疆哈萨克族传统发酵酸驼乳中分离的23株乳酸菌中,筛选出9株对金黄色葡萄球菌、枯草芽孢杆菌和大肠杆菌具有抑菌活性的菌株(其中茵株MLS5抑菌活性最强).通过有机酸、过氧化氢抑茵的排除试验和蛋白酶敏感性试验,初步确定MLS5产生的抑茵物质为蛋白质类细菌素.温度和pH耐受试验表明,该细菌素具有良好的热稳定性(121℃,20 min),pH 3～6具有抑茵活性.菌株MLS5的发酵曲线表明,菌株在37℃培养条件下具有良好的产细菌素和产酸能力,该细茵素在茵体生长对数中期产生,在生长稳定期抑茵活性达到最大.经抑茵谱试验表明,MLS5所产细菌素不仅能抑制革兰氏阳性细茵,而且对大肠杆菌和毛霉菌也有抑制作用,具有较广的抑茵谱.