实时荧光定量PCR法鉴定发酵乳中嗜热链球菌

该研究通过系统发育树分析确定目标基因,根据目的基因设计特异性引物和探针,建立一种能够快速准确鉴定发酵乳中嗜热链球菌的实时荧光定量聚合酶链式反应(RT-fqPCR)法,通过特异性、灵敏性和抗干扰实验对所建立方法进行验证,并使用该方法对市售的60份标识含有嗜热链球菌的发酵乳样品进行检测。结果表明,rec A基因具有种间特异性,种间差异率>10%,以其为目的基因建立的RT-fqPCR方法能够特异性的检测嗜热链球菌;绝对灵敏度达1 pg/μL,相对灵敏度达10³CFU/m L;在培养物水平和基因组水平抗干扰能力良好。采用该方法从60份标识含有嗜热链球菌的发酵乳样品中均能检测出嗜热链球菌,说明实时荧光定量PCR方法能够快速、准确的对发酵乳中嗜热链球菌进行检测。     

Lactobacillus delbrueckii subsp. bulgaricus和Streptococcus thermophilus接种比例对Bifidobacterium lactis益生菌发酵乳品质的影响

将Lactobacillus delbrueckii subsp.bulgaricus ND02(LB-ND02)和Streptococcus thermophilus ND03(ST-ND03)按1∶1、1∶10、1∶100、1∶1000接种于脱脂乳中,同时接入益生菌Bifidobacterium lactis V9(B.lactis V9,接种量为2.0×107g-1),于42℃进行发酵。通过对发酵及贮藏过程中发酵乳指标的测定,评价LB-ND02和ST-ND03的接种比例对发酵乳品质的影响。结果表明,随着LB-ND02接种比例减小,凝乳时间显著延长,B.lactis V9活菌数显著提高。4℃贮藏28 d后,随LB-ND02接种比例减小,B.lactis V9存活率差异显著,后酸化也显著减弱。研究发现,LB-ND02和ST-ND03的接种比例,显著影响发酵乳的发酵时间、B.lactis V9活菌数、后酸化及黏度。     

Effect of fermentation temperature on the properties of exopolysaccharides and the acid gelation behavior for milk fermented by Streptococcus thermophilus strains DGCC7785 and St-143

We investigated the effect of fermenting milk with 2 strains (DGCC7785 and St-143) of Streptococcus thermophilus, which are known to produce different types of exopolysaccharide (EPS) structures. The yields and physical properties of these ropy EPS were monitored during the fermentation of milk at different temperatures. We wanted to understand how these types of EPS properties affected yogurt gelation. Reconstituted skim milk was fermented at 33, 39, or 45°C until pH values reached 5.2, 4.9, 4.7, and 4.5. Molar mass of ropy EPS samples was determined using size exclusion chromatography coupled with multiangle laser light scattering. Rheological properties of fermented milk gels were analyzed using small-strain dynamic oscillatory measurements. In both strains, concentrations of ropy EPS increased during fermentation and at all temperatures. Fermentation times, by both strains, were shortest at 45°C and longest at 33°C. For both strains, molar mass of ropy EPS ranged from 2 to 4 × 10⁶ g/mol during fermentation. A major proteinaceous contaminant that was co-isolated with the ropy EPS fraction by our isolation method was identified as a milk-derived phosphoglycoprotein PP3. Increase in fermentation temperature from 33 to 45°C significantly decreased the storage modulus values (from 170 to 41 Pa) for milk gelled by strain DGCC7785, whereas the gels made with St-143 had very low storage modulus values (11–17 Pa) regardless of fermentation temperatures. For both strains, the values of maximum loss tangent in the milk gels increased with fermentation temperature; the maximum loss tangent occurred at higher pH values when milk was fermented by strain DGCC7785. The specific type of EPS produced appeared to be responsible for the differences in yogurt texture rather than the concentration or molar mass of the EPS.     

凝固型发酵豆浆的研制

以大豆作为原料,经混合乳酸菌发酵制备活性乳酸菌保健饮品。按照正交试验方案,确定了该产品制作的主要工艺参数为豆水比1∶10、接种量4%、培养温度40℃、发酵3h,产品口感细腻、香味浓郁。     

发酵黄芪粉对奶牛产奶量及乳成分的影响研究

研究发酵黄芪粉对奶牛产奶量及乳品质的影响.对照组奶牛饲喂基础日粮,试验Ⅰ、Ⅱ和Ⅲ组奶牛分别饲喂10、20和30 g/(头·d)的发酵黄芪粉基础日粮.结果表明:与对照组相比,试验组奶牛产奶量增加了0.95、1.45和1.58 kg/(头·d),但试验Ⅱ、Ⅲ组组间差异不显著;可显著提高奶牛乳蛋白率,但对奶牛乳脂率、乳糖及非脂固型物均无显著影响;可降低奶牛乳体细胞数,其中试验Ⅱ、Ⅲ组奶牛乳体细胞数显著低于对照组和试验Ⅰ组,但试验Ⅱ、Ⅲ组组间差异不显著;可提高奶牛经济效益,各试验组奶牛经济效益分别高于对照组2.6、3.4和2.72元/(头·d).该试验条件下,发酵黄芪粉以20 g/(头·d)的添加量较好.     

嗜酸乳杆菌发酵乳保藏性的研究

对嗜酸乳杆菌发酵乳在冷藏条件下保藏性进行了研究. 结果表明, 7 ℃条件下 14 d两株嗜酸乳杆菌发酵乳中细菌的存活率分别为 3.43% 和 2.20%, 且活菌数仍在 108 mL 1以上, 酸度为 90 ° T和 107 ° T, 双乙酰分别增加了 11.80% 和 56.41%, 乙醛含量分别为 2.993 mmol/ mL和 2.561 mmol/ mL, 使发酵乳具有良好风味.     

益生元在发酵乳中的应用研究进展

益生元是一种不可消化的食品成分,它能够调节肠道菌群促进人体健康。将益生元添加于发酵乳中能影响发酵乳的品质,益生元还可基于自身的益生特性增强发酵乳的功能性。本文主要针对益生元在发酵乳中应用研究进行综述,主要介绍了常见的低聚糖类益生元和多糖类益生元的性质,并分析了它们对于发酵乳的活菌数、产酸与风味、流变学性质和益生性评价等方面的作用效果,表明在发酵乳中添加益生元是益生元应用研究的重要方向。      

酶解对霉菌发酵乳风味的影响

以霉菌发酵乳为原料,以游离脂肪酸含量和游离氨基酸含量为检测指标,研究脂肪酶和蛋白酶的酶添加量、酶解温度、酶解时间3个因素对发酵乳风味的影响。结果表明,最优酶解条件为脂肪酶添加量1.5%,酶解温度45 ℃,酶解时间5 h,在此条件下游离脂肪酸含量为8.17%;蛋白酶添加量0.15%,酶解温度55 ℃,酶解时间5.5 h,在此条件下,游离氨基酸含量为0.41%。说明酶解可有效增加霉菌发酵乳风味物质,为进一步增强牛乳风味的研究与应用提供了基础。     

Effect of epigallocatechin gallate on the fermentative and physicochemical properties of fermented milk

Yogurt, a traditional fermented dairy product, is made with a starter that contains Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. The aim of this study was to investigate the effects of different concentrations of epigallocatechin gallate (EGCG; 0, 0.5, 1.0, 3.0, and 5.0 mg/mL) on the growth, metabolism, and acid production of lactic acid bacteria, as well as the texture, stability, and antioxidant activity of fermented milk (yogurt). The results showed that a low concentration of EGCG had no significant effect on the acid production capacity of the starter or on the water-holding capacity of the yogurt but did increase its viscosity. A high concentration (5.0 mg/mL) of EGCG delayed the acid production rate of the starter and decreased the water-holding capacity, but significantly increased the antioxidant activity of yogurt. The addition of EGCG significantly increased the hardness of yogurt. Therefore, EGCG can improve the texture of fermented milk and enhance its antioxidant activity and stability, thus improving the overall quality of yogurt.     

乳脂乳球菌复配发酵乳贮藏期品质变化研究

探讨了分离自新疆酸奶疙瘩的一株乳脂乳球菌,在乳制品应用中具有的潜在益生价值。通过测定多肽、滴定酸度、丁二酮、乙醛、黏度、活菌数等指标,研究了乳脂乳球菌复配菌株发酵乳在贮藏期间的发酵特性。结果表明,在贮藏期发酵乳各指标均显著高于对照组,酸度升高了29.59 °T,后酸化能力较强。丁二酮含量在整个贮藏过程中较稳定,乙醛、多肽、黏度、胞外多糖的含量在贮藏7 d后达到最大值。多肽含量最高达3.34 mg/mL,胞外多糖含量与测定的黏度值呈现正相关。活菌数呈下降趋势,贮藏21 d后,为7.8×107 CFU/mL,仍高于发酵乳标准中活菌数1×106 CFU/mL的要求。     

Replacement of bovine milk with vegetable milk: Effects on the survival of probiotics and rheological and physicochemical properties of frozen fermented dessert

In this study, frozen fermented desserts containing Lactobacillus acidophilus (La‐05; L) and Bifidobacterium bifidum (Bb‐12; B), were made from bovine (W), soya (S), coconut (C) and composite (i.e. combinations of coconut or bovine milks with soya milk) milks. The changes in frozen dessert eating qualities and the survival of added microbes were evaluated. The highest viscosity and melting resistance, and the lowest total sensory scores, were found in the products made using soya milk. After 90‐day storage at ?20 °C, the highest survival percentage of La‐05 was found in the products made using coconut milk (CL); and for Bb‐12, it was found in the products made using soya milk, coconut milk and a 25:75 blend of soya milk and coconut milk respectively ( samples SB, CB and SC3B) SB, SCB and CB samples.     

Metabolomic profile of milk fermented with Streptococcus thermophilus cocultured with Bifidobacterium animalis ssp. lactis,Lactiplantibacillus plantarum,or both during storage

In this study, the microbial interactions among cocultures of Streptococcus thermophilus (St) with potential probiotics of Bifidobacterium animalis ssp. lactis (Ba) and Lactiplantibacillus plantarum (Lp) in fermented milk were investigated during a storage period of 21 d at 4°C, in terms of acidifying activity (pH and titratable acidity), viable counts, and metabolites. A nontargeted metabolomics approach based on ultra-high-performance liquid chromatography coupled with mass spectrometry was employed for mapping the global metabolite profiles of fermented milk. Probiotic strains cocultured with St accelerated milk acidification, and improved the microbial viability compared with the single culture of St. The St–Ba/Lp treatment manifested a higher bacteria viability and acidification ability in comparison with the St–Ba or the St–Lp treatment. Relative quantitation of 179 significant metabolites was identified, including nucleosides, AA, short peptides, organic acids, lipid derivatives, carbohydrates, carbonyl compounds, and compounds related to energy metabolism. The principal component analysis indicated that St treatment and coculture treatments displayed a complete distinction in metabolite profiles, and Lp had a larger effect than Ba on metabolic profiles of fermented milk produced by cofermentation with St during storage. The heat map in combination with hierarchical cluster analysis showed that the abundance of metabolites significantly varied with the starter cultures over the storage, and high abundance of metabolites was observed in either St or coculture samples. The St-Ba/Lp treatment showed relatively high abundance for the vast majority of metabolites. These findings suggest that the profile of the metabolites characterizing fermented milk samples may depend on the starter cultures, and incorporation of probiotics may considerably influence the metabolomic activities of fermented milks.     

快速功能性嗜酸乳杆菌发酵乳的研制

对功能性嗜酸乳杆菌和嗜热链球菌混合菌种发酵乳的最佳生产工艺条件进行研究。结果表明:嗜酸乳杆菌和嗜热链球菌以1∶1配合、添加6%的蔗糖和适量的蛋白糖、9%的接种量、41℃5h培养生产出具有酸奶特有的质地和风味,嗜酸乳杆菌活菌数为2.9×107cfu/mL,保藏期为12d的功能性发酵乳。与单一嗜酸乳杆菌发酵乳的生产周期18h相比,缩短了13h,且风味更佳。     

芡实发酵奶发酵工艺优化研究

基于保加利亚乳杆菌和乳酸链球菌制作芡实发酵奶工艺,在单因素实验基础上选取接种量、发酵时间、发酵温度、柠檬酸添加量为自变量,利用响应面分析法研究各自变量及交互作用对芡实发酵奶中乳酸菌浓度的影响,模拟得到二次多项式回归方程的预测模型。实验结果表明,接种量为3.3%,发酵时间为7.4h,发酵温度为40.5℃,柠檬酸添加量为0.16%时乳酸菌浓度最高,在此条件下乳酸菌浓度平均值为1.923×109cfu/mL,与理论预测值1.920×109cfu/mL相比相对误差为1.56%,说明通过响应面优化得出的回归方程有一定的实践指导意义。      

嗜酸乳杆菌生物学特性及其发酵乳的研究

研究了嗜酸乳杆菌的生物学特性,并确定其发酵乳的合适的加工工艺。结果表明,在质量分数为11％脱脂乳培养基中加入质量分数为7％的啤酒能显著提高嗜酸乳杆菌的产酸速率及活菌总数。该菌株耐盐和耐胆酸盐能力分别达到6 0％和1.5％,同化胆固醇达80％,证明是一株性能优良的益生菌株。在此基础上确定嗜酸乳杆菌发酵乳的加工工艺:①采用乳糖部分水解的牛乳（水解率40％）为原料,接种质量分数为3％的嗜酸乳杆菌;②采用混合菌种（嗜酸乳杆菌+唾液链球菌嗜热亚种+德氏乳杆菌保加利亚亚种=3％+0.5％+0.5％）为发酵剂,按常规酸乳工艺生产。产品中嗜酸乳杆菌的活菌数达到109mL-1以上,5d后其活菌数仍有6×108mL-1,且产品口风味良好。     

高效液相色谱法测定混菌发酵乳中乳糖、葡萄糖及半乳糖含量的变化

利用高效液相色谱法测定混菌发酵乳中乳糖、葡萄糖和半乳糖含量及其变化。样品经TCA沉淀蛋白,采用AminexHPX-87C色谱柱,超纯水为流动相,流速0.3mL/min,使用示差折光检测器进行检测。结果表明:利用该方法进行色谱分析,各组分能够达到很好的分离,各糖组分在0.5～10mg/mL范围内呈现良好的线性关系,相关系数均达到0.9999以上,检出限为0.012～0.016mg/mL,样品加标平均回收率为91.30%～103.28%,精密度实验表明,相对标准偏差在0.49%～3.82%之间（n=6）,该法用于实际样品的测定可靠、准确。      

光谱法测定发酵乳中乙醛和双乙酰含量的变化

为了考察乳酸菌单菌种发酵乳的产酸特性和产香特性。使用酚试荆分光光度法测定发酵乳中乙醛含量，使用邻苯二胺法测定发酵乳中的双乙酰含量，并结合滴定酸度法测定发酵乳中的酸度。研究了L．b-3，Sf-9和L.dia在不同温度下发酵过程中的乙醛、双乙酰含量和酸度变化，以此评估发酵乳中有机酸和羰基化舍物的情况。     

嗜酸乳杆菌在现代乳品中的应用研究进展

分析了现代乳品生产状况及存在问题，概述了嗜酸乳杆菌生物学特性、生理功能以及在乳品工业上的应用及发展趋势。     

乳酸菌与酵母菌对牦牛酸乳品质形成的影响

为明确乳酸菌和酵母菌在牦牛酸乳品质形成中发挥的作用,以牦牛乳粉为原料、牦牛酸乳为发酵剂发酵酸乳,在发酵过程中分别抑制乳酸菌和酵母菌的活性,测定并比较正常发酵、抑制乳酸菌发酵、抑制酵母菌发酵酸乳的风味、口感、质构等相关指标。结果表明,乳酸菌和酵母菌对牦牛酸乳品质形成均起到关键作用,乳酸菌对酸乳酸度、质构、氨基酸态氮、有机酸、V_(B1)及酮类和酸类风味物质的贡献较大,酵母菌对乙醇、V_(B2)及醇类和酯类风味物质的贡献较大。     

Effects of milk products fermented by Bifidobacterium longum on blood lipids in rats and healthy adult male volunteers

The effects of milk products fermented by Bifidobacterium longum strain BL1, a probiotic strain, on blood lipids in rats and humans were studied. Rats were fed a cholesterol-enriched experimental diet, supplemented with lyophilized powders of 1) acid milk (control), 2) milk fermented with a mixed culture of ordinary yogurt starters composed of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (SL), and 3) bifidobacterium milk fermented with the probiotic B. longum strain BL1, respectively. The bifidobacterium milk feeding brought about significant lowering of the serum concentrations of total cholesterol, low-density lipoprotein cholesterol, and triglycerides, in comparison with the control, while no change in high-density lipoprotein cholesterol concentration was observed. On the other hand, supplementation with SL milk resulted in only slight, nonsignificant decreases in serum lipid concentrations in comparison with the control. In the human study, 32 subjects with serum total cholesterol ranging from 220 to 280 mg/dl were randomly assigned to two treatments: 1) intake of a low-fat drinking yogurt prepared with ordinary yogurt starters composed of S. thermophilus and L. delbrueckii subsp. bulgaricus (P-group) and 2) intake of a low-fat drinking yogurt prepared with the two ordinary yogurt starters plus B. longum strain BL1 (B-group). After intake for 4 wk at 3 x 100 ml/day, reduction of serum total cholesterol was observed in approximately half of the B-group subjects; a particularly significant decrease in serum total cholesterol was found among subjects with moderate hypercholesterolemia (serum total cholesterol > 240 mg/dl). However, the serum lipid concentrations in the P-group subjects were almost stable during the experimental periods. The present results indicate the potential of the probiotic B. longum strain BL1 in serum lipid improvement.