乳酸菌发酵酸豆乳香气成分检测分析及评价

采用顶空固相微萃取(SPME)与气相色谱- 质谱(GC-MS)联用检测分析技术,并结合感官评分法对保加利亚乳杆菌、瑞士乳杆菌和嗜热链球菌3 种乳酸菌混合发酵的豆乳进行香气分析和评价,探讨不同乳酸杆菌及其与嗜热链球菌按比例混合发酵对酸豆乳香气的影响。感官评定结果表明,乳酸菌种及其混合比例对发酵酸豆乳的香气有明显影响。香气检测分析发现,对比发酵前样品,发酵后酸豆乳中酮类、酸类含量显著增加,而醛类和醇类含量明显减少,3 种酸豆乳中相同关键香气成分含量有差异。感官评定与香气检测分析结果一致,1:1 的保加利亚乳杆菌和嗜热链球菌组成的菌种产香浓郁,产酸力高,是豆乳发酵的优选菌种。     

Development of yoghurt from ovine milk with enhanced texture and flavour properties

Six commercial starter cultures, all blends of Streptococcus thermophilus sp. and Lactobacillus bulgaricus sp., were compared for enhanced texture and flavour in yoghurt prepared from ovine milk. The fermentation patterns differed between starters, with YF‐L903 (E) showing the fastest fermentation but slower post‐acidification. YC‐X11 (A) produced the slowest acidification, while YC‐380 (B) resulted in the fastest post‐acidification. Textural and colour properties were affected significantly by the culture used. YF‐L903 (E) enhanced the firmness and luminosity of the yoghurt over storage, increasing mouthfeel and creaminess, as compared to yoghurts prepared with the other starter cultures starters studied.     

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。     

Proteolytic degradation of ewe milk proteins during fermentation of yoghurts and storage

Yoghurts are mostly produced from cow milk and to a very limited extent from ewe milk. The evolution of caseins and whey proteins in ovine milk submitted to different thermal treatments (63 degrees C/30 min; 73 degrees C/15 min; 85 degrees C/10 min or 96 degrees C/5 min) was followed during fermentation of yoghurts and during their storage up to 14 days, using two different sets of starters. One set of starter LAB was a "ropy" culture (YC-191), which is a well-defined mixed strain culture containing Streptococcus thermophilus ST-143 and Lactobacillus delbrueckii subsp. bulgaricus (LB-18 and LB-CH2). The other set of starter bacteria (YC-460) was a standard yoghurt culture("non-ropy") containing mixed strain culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. Contents of free amino groups in produced yoghurts increased gradually during the fermentation, up to a maximal value obtained after 4 h fermentation, then they did not change significantly during storage of yoghurt produced with YC-191 starter. In contrary, a large drop in the amount of free amino groups was observed in the first 24 h of storage in the case of yoghurt made with YC-460 indicating that microorganisms continue still to grow in low temperatures. During fermentation and storage of both yoghurt types, alpha-lactalbumin was hydrolyzed to a slightly bigger extent than beta-lactoglobulin. During fermentation, beta-casein was slightly more degraded than alpha(s)-caseins; however, the opposite was observed during storage up to 14 days. Generally, a more intense heat pretreatment led to a higher degradation of whey proteins and caseins during fermentation and storage. Differences in proteolytic activity between the two starters used (whey proteins more degraded by YC-191; caseins more degraded by YC-460) may lead to improvement in production and formulation of yoghurts differing in their physicochemical and rheological properties.     

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。     

Effect of thermophilic lactic acid bacteria on the viability of Salmonella serovar Typhimurium PT8 during milk fermentation and preparation of buffalo's yogurt

Viability of dairy-borne Salmonella enterica ssp. enterica serovar Typhimurium PT8 was studied during the fermentation of skim milk by thermophilic lactic acid bacteria (LAB). Longer generation times of Salmonella were found in mixed cultures of skim milk containing Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus or a mixture of them (1:1), as compared with single cultures of the pathogen. Salmonella was less able to survive in mixed cultures with these LAB during prolonged incubation at 41°C and also during cold storage of the fermented milk. L actobacillus ssp. bulgaricus and its mixture with S. thermophilus were more inhibitory to the growth and survival of Salmonella than was S. thermophilus . This was associated with higher ability of L . ssp. bulgaricus and the mixture to develop acidity in milk than S. thermophilus . Examining the antibacterial activity of these LAB towards Salmonella showed that other factors including heat-resistant and heat-labile compounds were involved in inhibiting the pathogen by these cultures. The viability of the same Salmonella strain during the preparation and cold storage of buffalo's yogurt was also examined. Salmonella was found to survive longer in yogurt made with starter containing probiotic bacteria than in that prepared with the traditional starter. This was ascribed to the development of lower pH by the traditional starter.     

不同乳酸菌对凝固型荔枝酸奶的发酵特性和质构的影响

以纯牛奶和荔枝汁为主要原料,以干酪乳杆菌、保加利亚乳杆菌和嗜热链球菌为发酵菌种,探讨三种乳酸菌的不同组合发酵对凝固型荔枝酸奶的发酵特性和质构的影响。结果表明:干酪乳杆菌产胞外多糖能力较强,单独发酵时胞外多糖含量达到22.50g/L,而嗜热链球菌和保加利亚乳杆菌单独发酵时胞外多糖的含量分别为5.73 g/L和0.29 g/L;各种不同乳酸菌组合发酵后酸奶的表观粘度和持水力与胞外多糖的含量呈正相关(R2=0.98);干酪乳杆菌产酸能力弱,单独发酵后p H高于其它添加有嗜热链球菌和保加利亚乳杆菌的组合,导致酸奶的硬度偏低,但干酪乳杆菌联合嗜热链球菌或保加利亚乳杆菌发酵时,发酵酸奶的硬度和乳酸菌活菌数均明显优于单独发酵组。因此,当干酪乳杆菌与嗜热链球菌及保加利亚乳杆菌联合发酵时,能充分发挥三个菌种的各自优势,菌落总数、胞外多糖含量和质构均能达到较好的品质水平。     

大豆蛋白酶解产物对不同发酵剂酸奶品质的影响

本文研究了大豆蛋白酶解产物(Soy protein hydrolysate,SPH)对187、495和885三种发酵剂酸奶发酵过程中p H值变化和贮藏过程中乳清析出、流变特性和质构特性的影响。结果表明,SPH对3种酸奶均具有一定的促发酵效果,其中对以187和495为发酵剂的p H值下降速度较慢的酸奶促发酵效果更明显。贮藏过程中,SPH可有效减缓酸奶乳清析出,其中对以885为发酵剂的乳清析出率最高的酸奶作用最明显,降低率达到49.35%。此外,SPH可有效提高187和495发酵剂酸奶的表观黏度和触变环面积,有利于酸奶结构的恢复。通过对酸奶第7 d流变和质构特性的进一步研究,SPH可显著提高3种酸奶的稠度和零剪切黏度,降低其流动指数、柔量和粘性指数,从而有效改善3种酸奶的流变和质构特性,提高酸奶在低应力下稳定性,降低其在高应力下形变破坏程度。     

Yoghurt fermented by Lactobacillus delbrueckii subsp. bulgaricus H+ -ATPase-defective mutants exhibits enhanced viability of Bifidobacterium breve during storage

Persistent acid production by Lactobacillus delbrueckii subsp. bulgaricus during refrigerated storage is a major cause of reduced viability of probiotic strains such as Bifidobacterium breve in yoghurt. It was established that H+ -ATPase-defective mutants of lactic acid bacteria have reduced growth and metabolism in low pH environments. Therefore, the aim of this study was to evaluate inhibition of post-acidification and maintenance of B. breve viability in yoghurt fermented by L. delbrueckii subsp. bulgaricus mutants with reduced membrane-bound H+ -ATPase activity during refrigerated storage. Spontaneous neomycin mutants of L. delbrueckii subsp. bulgaricus that had a significantly (P < or = 0.05) reduced H+ -ATPase activity were successfully isolated. Yoghurt fermented using L. delbrueckii subsp. bulgaricus SBT0164 No. 55-1 (mutant) starter culture had markedly reduced post-acidification and maintained viability (> or = 10(8) CFU/ml) of both Bifidobacteruim breve JCM 1192(T) and Bifidobacteruim breve JCM 7017 during storage at 10 degrees C for 21 days. These results clearly showed that yoghurt fermented by mutants of L. delbrueckii subsp. bulgaricus with reduced membrane-bound H+ -ATPase activity has reduced post-acidification that prolongs viability of B. breve in yoghurt during refrigerated storage.     

Effect of addition of amino acids, treatment with β-galactosidase and use of heat-shocked cultures on the acetaldehyde level in yoghurt

In this study, the biochemical activities of nonviscous and viscous yoghurt starter cultures were investigated. Yoghurt samples produced with nonviscous and viscous cultures, and viscous cultures + methionine (10 and 30 mg/100 mL milk), + threonine (5 and 10 mg/100 mL milk), + β-galactosidase (1 mg/100 mL milk), and with a heat-shocked culture were analysed. In the experimental yoghurts, the pH, titratable acidity, lactic acid, tyrosine and acetaldehyde contents and the number of total starter organisms were determined. According to the results obtained, the samples produced with the viscous culture had the lowest acetaldehyde levels, whereas the highest acetaldehyde level was found in the samples manufactured with the nonviscous culture. Compared with the samples inoculated with the viscous culture alone, the amino acid supplementation, lactose hydrolysis and heat-shock treatments caused a significant increase in the level of acetaldehyde.     

新疆传统乳制品中优良乳酸菌的筛选及多菌株发酵剂的研究

从新疆地区采集的46份传统乳制品中分离获得的612株乳酸菌,根据菌株来源、培养条件,形态特征、凝乳状态和产酸特性,选择其中20株乳酸菌进行生理生化鉴定和酸奶发酵试验,结果表明,分离株中3株为链球菌,4株为乳球菌,13株为乳杆菌,平均发酵产酸速率在8.11~12.43oT/h,不同菌株在酸乳冷藏期间表现出不同的后酸化活性,冷藏15d滴定酸度提高了6.3%~55.7%。不同菌株凝乳时黏度在272~1085cp,多数菌株发酵乳在冷藏期间黏度呈上升趋势,球菌的产黏能力普遍低于杆菌。气相色谱分析结果显示不同菌株发酵乳中丁二酮在3.51~68.74mg/L,乙醛在3.51~68.74mg/L,丙酮在0.87~48.35mg/L,乙酸乙酯在0~103.68mg/L,异戊醇都未检出。经过混合菌株发酵生产酸乳比较,发现以S.thermophilusX1、S.thermophilusX5、L.plantarumY5和L.delbrueckii bulgaricussubsp.Y10按1∶1∶1∶1比例组合作为发酵剂,生产的酸乳组织状态较好、风味独特、后酸化弱。     

Effect of milk base and starter culture on acidification,texture, and probiotic cell counts in fermented milk processing

In the present work, the compared effect of milk base and starter culture on acidification, texture, growth, and stability of probiotic bacteria in fermented milk processing, was studied. Two strains of probiotic bacteria were used, Lactobacillus acidophilus LA5 and L. rhamnosus LR35, with two starter cultures. One starter culture consisted only of Streptococcus thermophilus ST7 (single starter culture); the other was a yogurt mixed culture with S. thermophilus ST7 and L. bulgaricus LB12 (mixed starter culture). For the milk base preparation, four commercial dairy ingredients were tested (two milk protein concentrates and two casein hydrolysates). The resulting fermented milks were compared to those obtained with control milk (without enrichment) and milk added with skim milk powder. The performance of the two probiotic strains were opposite. L. acidophilus LA5 grew well on milk but showed a poor stability during storage. L. rhamnosus LR35 grew weakly on milk but was remarkably stable during storage. With the strains tested in this study, the use of the single starter culture and the addition of casein hydrolysate gave the best probiotic cell counts. The fermentation time was of about 11 h, and the probiotic level after five weeks of storage was greater than 106 cfu/ml for L. acidophilus LA5 and 10(7) cfu/ml for L. rhamnosus LR35. However, an optimization of the level of casein hydrolysate added to milk base has to be done, in order to improve texture and flavor when using this dairy ingredient.     

酸奶发酵菌的研究及其风味化合物的鉴定

酸奶是一种营养价值很高,又富有良好风味的广为消费者喜爱的传统发酵乳制品。本文对酸奶发酵菌的研究和酸奶中主要风味化合物的分析鉴定进行了初步探讨。在发酵菌方面,选择了本所保藏的6038号(str. thermophilus)和6047号(Lb. bulgaricus)两支较好的乳酸菌为本实验的出发菌株。按1∶1的配合比,保温在43℃下进行牛乳发酵,凝结后立即放在4～7℃下保持24小时左右,可以获得较为满意的发酵乳风味。在挥发性风味化合物的分析鉴定方面,采用了真空蒸馏、溶剂提取和调节pH分离的方法,将风味组分分为酸类和非酸类两部分,经浓缩后使用气相色谱(GC)分析技术、气相色谱—质谱(GC—MS)联用技术和离子色谱(IC)分析技术,分析鉴定了酸奶中主要风味化合物;并定性定量分析了不同发酵和低温保持阶段上重要香气成份——双乙酰的生物合成水平。由此得到结论。发酵培养完成后,于低温保持24小时,酸奶中双乙酰的生成量最高可达到10.34ppm。该水平双乙酰含量可以赋与酸奶特有的香气和良好的风味。进而推论,此含量范围的双乙酰应为酸奶中的较适水平;酸奶发酵进行后,低温保持阶段的初期对酸奶风味的形成有十分重要的意义。     

大豆酸乳发酵菌种的驯化及营养因子的研究

选择保加利亚杆菌（Lactobacillus bulgaricus）和嗜热链球菌（Streptococcus thermophilus）作为发酵菌种,通过逐步提高牛奶中豆乳的比例对菌种进行驯化的方法和改变培养基中营养成分的含量,研究大豆酸乳发酵过程中酸度值和菌落总数的变化情况。结果表明,经过驯化后的保加利亚杆菌发酵大豆酸乳过程中,酸度值从37.5 °T上升至40.0 °T,活菌数量提高了0.5倍;经过驯化后的嗜热链球菌发酵大豆酸乳过程中,酸度值从35.0 °T上升至37.5 °T,活菌数量提高了1.67倍;经过驯化后的混合菌种（保加利亚杆菌∶嗜热链球菌比例为1∶1）发酵大豆酸乳过程中,酸度值从42.5 °T升至48.5 °T,活菌数量提高了1.85倍;5种营养因子对大豆酸乳酸度的提高作用顺序为大豆肽＞氨基酸＞胡萝卜汁＞低聚果糖＞西红柿汁。     

Effect of probiotic cultures on the stability of anthocyanins in blueberry yoghurts

Dependent on anthocyanin pigments, colour is one of the main quality factors of berry products. In this paper we assess the influence of probiotic bacteria on the degradation rate of anthocyanins in yoghurts with a highbush blueberry preparation during storage. Four types of yoghurts were prepared: first with the yoghurt starter culture YC-X16 only (Steptococcus thermophilus and Lactobacilllus delbrueckii subsp. bulgaricus) and three more, each with the starter culture YC-X16 and one of the three probiotic cultures: Bb-12, La-5 and LCP, respectively (Bif. animalis subsp. lactis – Bb-12; Lactobacillus. acidophilus – La-5 and Lactobacillus. paracasei subsp. paracasei – LCP).Derivatives of malvidin were the predominant anthocyanins in the tested yoghurts. Degradation of pigments occurred in accordance with the first-order reaction and its half-life time depended on the bacterial cultures. Anthocyanins in the probiotic yoghurt made with LCP culture were characterized by lower stability than those made with the other bacterial cultures. The impact of lactic bacteria or their metabolic products on the stability of anthocyanins in berry yoghurts shows that an appropriate selection of culture for production of yoghurt is recommended.     

Interactions between EPS-producing Streptococcus thermophilus strains in mixed yoghurt cultures

Mixed cultures of different EPS-producing Streptococcus thermophilus strains in combination with a Lactobacillus delbrueckii subsp. bulgaricus strain with negligible EPS-production were used for yoghurt production. The yoghurt texture was characterised with respect to sensory, rheological and microstructural properties and the EPS-concentrations were determined. The cultures resulted in yoghurts with highly different texture properties, and positive interactions between certain Streptococcus thermophilus strains were observed. The underlying properties of yoghurt texture are multidimensional, but a number of microstructural characteristics were apparent in the yoghurts with the highest mouth thickness, creaminess and viscosity. A strong protein network, not too dense and with medium size pores containing EPS, seems associated with these properties. The presence of capsular polysaccharides (CPS) also appeared to be beneficial as did a combination of EPS types, which were distributed differently in the protein network (in serum pores, respectively in association with protein). Obviously, a certain concentration of EPS must be present to provide for these effects on yoghurt texture, but other factors than concentration per se seem more important.     

Physical characteristics of yoghurts made using exopolysaccharide-producing starter cultures and varying casein to whey protein ratios

This study investigated the physical characteristics of set and stirred yoghurts made at 9% (w/w) total solids with various casein (CN)-to-whey protein (WP) ratios and with exopolysaccharide (EPS)-producing starter cultures (capsular or ropy) during storage. The yoghurt was evaluated for composition, firmness and syneresis of set yoghurt, and for the flow curve and the area of hysteresis loop between the upward and downward curve of stirred yoghurt. Viable counts of starter bacteria and concentrations of lactic acid and EPS in the yoghurt were also determined. EPS concentration did not decrease during storage for 28 d. Firmness and syneresis of set yoghurt decreased when the CN-to-WP ratio was reduced from ratio 4:1 to 1:1 and when EPS starter cultures (especially ropy EPS) were used. Stirred yoghurt with a CN-to-WP ratio of 3:1 and made using ropy EPS-producing starter cultures had a higher shear stress and hysteresis loop area than yoghurt made using capsular EPS- or non-EPS-producing starter cultures. The results suggested that the physical characteristics of set and stirred yoghurts can be improved by varying CN-to-WP ratio and by the use of EPS-producing starter cultures.     

罗地亚9001直投型发酵剂发酵特性研究

利用罗地亚90001直投型发酵剂来发酵酸奶，探讨了利用直投型发酵剂发酵的酸奶不能作为后续发酵剂进行酸奶生产的原因。在实验中首先利用直投型发酵剂发酵牛乳，然后依次利用各代次产物作为后续发酵剂生产酸奶，直至第6代牛乳不能再正常发酵为止。同时，对每代酸奶中嗜热链球菌和保加利亚乳杆菌进行计数，并观察嗜热链球菌和保加利亚乳杆菌比例上的变化。最后将第6代不能正常发酵酸奶中的嗜热链球菌和保加利亚乳杆菌分离并进行鉴定。将二者按1：1(数量比)的比例接种牛乳，发现牛乳能够正常发酵。结果表明。利用酸奶作为后续发酵剂的第6代牛乳不能正常发酵的原因是嗜热链球菌和保加利亚乳杆菌的数量比发生变化。而导致此情况发生的原因可能是由于低温下二者的生长速率不同。     

Viili直投式发酵剂复配及发酵性能研究

以Viili发酵乳为研究对象,通过测定酸度、pH值、持水力、表观黏度、双乙酰含量及乙醛含量等理化指标,得到Viili发酵乳的最适发酵时间为12 h,此时pH值为4.69,酸度为91.85 °T,持水力为51.78%,表观黏度为4 457 mPa·s,双乙酰含量为15.67 mg/L,乙醛含量为10.63 mg/L。确定最优复配发酵V7组菌相为嗜热链球菌（Streptococcus thermophilus）、保加利亚乳杆菌（Lactobacillus bulgaricus）、乳酸乳球菌乳脂亚属（Lactococcus lactis subsp. cremoris）、假肠膜明串珠菌（Leuconostoc pseudomesenteroides）、白地霉（Galactomyces candidum）与克鲁维毕赤酵母（Pichia kluyver）的比例为162∶16∶8∶14∶74∶1,凝乳时间为19 h,酸度为74.72 °T,持水力为38.10%,表观黏度为7 780 mPa·s,双乙酰含量为7.87 mg/L,乙醛含量为15.4 mg/L。复配发酵V7组与Viili发酵乳12 h的理化性质整体接近,具备Viili发酵乳的特色品质及风味物质,为后续Viili发酵乳直投式发酵剂产品的开发提供理论基础。     

Distribution and stability of aflatoxin M1 during production and storage of yoghurt.

Yoghurt from cow's milk artificially contaminated with aflatoxin M1 (AFM1) at levels of 0.050 and 0.100 g l(-1) was fermented to reach pHs 4.0 and 4.6. Yoghurt fermented to pH 4.6 was also used for preparing strained yoghurt. Yoghurts were stored at 4 degrees C for up to 4 weeks. Analysis of AFM1 in milk, yoghurt, strained yoghurt and yoghurt whey was carried out using immunoaffinity column extraction and liquid chromatography coupled with fluorometric detection. AFM1 levels in yoghurt samples showed a significant decrease (p < 0.01) compared with those initially added to milk. Growth of culture lactic acid bacteria was not affected in the AFM1 contaminated yoghurts, with the exception of Streptococcus thermophilus that showed a significantly (p < 0.01) lower increase in the yoghurt containing the toxin at high concentration. Following fermentation, AFM1 was significantly lower (p < 0.01) in yoghurts with pH 4.0 than in yoghurts with pH 4.6 at both contamination levels. During refrigerated storage, AFM1 was rather more stable in yoghurts with pH 4.6 than with pH 4.0. The percentage loss of the initial amount of AFM1 in milk was estimated at about 13 and 22% by the end of the fermentation, and 16 and 34% by the end of storage for yoghurts with pHs 4.6 and 4.0, respectively. The percentage distribution ratio of AFM1 in strained yoghurt/yoghurt whey of the initial toxin present in the yoghurt was about 90/10 and 87/13 for the lower and the higher contamination levels, respectively.