SPME-GC-MS结合ROAV分析单菌及复配发酵牛乳中关键性风味物质

采用固相微萃取-气相色谱-质谱联用技术检测分析Streptococcus thermophilus与Lactobacillus delbrueckii subsp.bulgaricus单菌及复配发酵牛乳中的挥发性风味物质,结合相对气味活度值(relative odor activity value,ROAV)探讨发酵牛乳中关键性风味物质。结果表明:本实验共鉴定出100种挥发性风味物质,包括酸类、酮类、醛类、醇类、酯类、烷烃类和芳香族类化合物等。主成分分析表明,表征S.thermophilus单菌发酵乳的关键性风味物质是双乙酰、正壬醛和甲苯;表征L.bulgaricus单菌发酵乳的关键性风味物质是正庚醛、丁酸-2-甲基丙酯和1-庚醇;表征S.thermophilus与L.bulgaricus复配发酵乳的关键性风味物质是乙醛、3-甲基正丁醛、乙偶姻、2-壬酮、2-庚酮、醋酸乙烯酯、碳酸庚基苯基酯、甲酸乙烯酯和2-壬醇。相较于单菌发酵,复配发酵的风味物质组成、各组分相对含量及关键性风味物质均发生改变。     

优良“老酸奶”发酵剂组合的筛选

为了筛选出优良的酸奶发酵剂,根据菌株的稳定性、发酵活力、凝乳时间等指标对23菌株(其中有13株球菌与10株杆菌)进行初步筛选,获得13株具有优良发酵特性的菌株(其中有7株球菌与6株杆菌)。根据单菌株发酵实验结果,按菌种性能互补原则将各球菌与杆菌复配,进行28个组合发酵实验,测定发酵速度、质构、乳清析出以及后酸化等指标。最终筛选出球菌S1、S4与杆菌L2,此组合与目前商业化直投发酵剂比具有发酵前期产酸速度快、发酵时间短的优点,而且具有天然的发酵香气,乳清析出量很少,适用于直投式发酵剂的生产。     

EFFECT OF COLD STORAGE ON CULTURE VIABILITY AND SOME RHEOLOGICAL PROPERTIES OF FERMENTED MILK PREPARED WITH YOGURT AND PROBIOTIC BACTERIA

ABSTRACT

We examined the effect of storage time on culture viability and some rheological properties (yield stress, storage modulus, loss modulus, linear viscoelastic region, structural recuperation and firmness) of fermented milk made with Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus acidophilus (LA) and Bifidobacterium animalis ssp. lactis in coculture with Streptococcus thermophilus (ST). Acidification profiles and factors that affect viability (postfermentation acidification, acidity and dissolved oxygen) were also studied during 35 days at 4C. Fermented milk prepared with a coculture of ST and Bifidobacterium lactis gave the most constant rheological behavior and the best cell viability during cold storage; it was superior to ST plus LA for probiotic fermented milk production.

PRACTICAL APPLICATIONS

Probiotic cultures should grow quickly in milk, provide adequate sensory and rheological properties to the product, and remain viable during storage. Commercially, it is very common to use yogurt starter culture (i.e. Streptococcus thermophilus[ST] and Lactobacillus delbrueckii ssp. bulgaricus) in combination with the probiotic bacteria in order to reduce fermentation time. However, LB tends to post acidify fermented milk, which reduces the viability of the probiotic bacteria; thus, it is recommended to use starter cultures devoid of this species. We found that the technological properties and the viability of the probiotic bacterium Bifidobacterium animalis ssp. lactis BL O4 in coculture with ST make it suitable for probiotic fermented milk production; it produces rheological characteristics similar to those of yogurt.     

Characterization of volatile compounds in fermented milk using solid-phase microextraction methods coupled with gas chromatography-mass spectrometry

Lactic acid bacteria (LAB) are industrially important bacteria that are widely used in the fermented food industry, especially in the manufacture of yogurt. Characteristic flavors are produced by LAB during fermentation and storage that affect the quality and acceptability of fermented milk products. In this study, the volatile compounds in milk fermented by Streptococcus thermophilus IMAU80842 alone, Lactobacillus delbrueckii ssp. bulgaricus IMAU20401 alone, or both species together were identified using solid-phase microextraction methods coupled with gas chromatography-mass spectrometry. A total of 53, 43, and 32 volatile compounds were identified in milk fermented by S. thermophilus alone, L. delbrueckii ssp. bulgaricus alone, or both species together, respectively. The presence of some important flavor compounds was confirmed: acetic acid, acetaldehyde, acetoin, 2,3-butanedione, ethanol, and 1-heptanol. Our results demonstrate that the composition of the volatile compounds in fermented milk depends on the species of LAB used and whether they are used alone or in combination. This is important for the selection of appropriate starter cultures for the production of different types of fermented milk product with particular flavors.     

Production of a yogurt-like product from plant foodstuffs and whey. Sensory evaluation and physical attributes

A mixed substrate composed of soya milk, oat flour and dried cheese whey (820, 110 and 70 g kg^?1 respectively) was heat treated (80°C, 20 min) and fermented using two different yogurt starters. Sensory evaluation was conducted in order to get the basic flavour profile and to assess the acceptability of the product. Unfermented mixed substrate and fermented milk were used as references. Two yogurt starter combinations were used. Some additives such as sugar and calcium were also assessed. The addition of an equal weight of milk to the mixed substrate, and flavours such as strawberry jam or honey, were tried as well. Acceptability of the mixed substrate was increased by fermentation and added sugar, milk and/or flavours. A suitable combination of strains was very important to get good acceptability of the fermented product. Colour and syneresis were also evaluated. Heat treatment had very little influence on the colour of the mixed substrate. The mixture was less white and a little less green than milk. Syneresis was lower than that of a yogurt made from milk with 145 g litre^?1 total solids.     

The application of staphylococci with flavour-generating potential is affected by acidification in fermented dry sausages

Differences in the production of bacterial metabolites with potential impact on fermented sausage flavour were found in meat simulation medium when comparing different strains of Staphylococcus xylosus and Staphylococcus carnosus as starter cultures. Overall, higher levels of 3-methyl-1-butanol and acetoin were found for S. xylosus, with some intraspecies variability. In addition, sausage fermentation parameters affected staphylococcal growth and metabolism. Strong acidification, as in Northern-European types of fermented dry sausage, inhibited S. xylosus 3PA6 but not S. carnosus 833. During a milder, Southern-European type of acidification, both strains displayed survival over time. During in situ sausage trials, variations in the degree of acidification and the choice of starter microorganisms were of importance, whereas modifications in fat and salt contents had no effects. Staphylococcus sciuri αSg2, Staphylococcus succinus 4PB1, and S. xylosus 3PA6 were unable to survive the fermentation of a Northern-European type of fermented dry sausage, characterized by low or no 3-methyl-1-butanol and acetoin production. Inoculation with S. sciuri αSg2, S. succinus 4PB1, or S. xylosus 3PA6 led to 3-methyl-1-butanol and acetoin production in Southern-European type of fermented dry sausages, which was not observed with S. carnosus 833.     

Rheological, sensorial, and chemopreventive properties of milk fermented with exopolysaccharide-producing lactic cultures

The objective of this research was to evaluate the rheological, sensorial, and chemopreventive properties of milk fermented with different exopolysaccharide (EPS)-producing lactic cultures. Reconstituted skim milk (11% wt/vol) was fermented with single strains of EPS-producing and non-EPS-producing cultures. Whey that collected on the surface of undisturbed fermented milks and after cutting was measured. All EPS-producing cultures reduced the amount of whey present on the surface of the undisturbed samples, whereas only 3 out of 5 strains reduced syneresis measured after cutting. All EPS-producing cultures except a strain of Lactobacillus delbrueckii ssp. bulgaricus reduced viscoelastic moduli in fermented milk. There was a linear correlation between ropiness and smoothness. In the chemoprevention study, 140 male Fisher rats were divided into 7 groups of 20 each. Rats in 6 groups were fed diets supplemented with fermented milks each made with a single strain of EPS-producing or non-producing cultures, whereas rats in group 7 (control) were fed a diet supplemented with milk acidified with glucono-δ-lactone (GDL). All rats were injected with azoxymethane (15 mg/kg, subcutaneous) at wk 7 and 8 of age to induce tumors and fed their respective diets ad libitum throughout the study. After 30 wk of initiation, all rats were anesthetized with ether, and their intestinal tissues were isolated and washed with cold normal saline. The number and size of tumors in the colon and small intestine were recorded. Rats fed diets supplemented with fermented milk made with 2 EPS-positive and 1 EPS-negative strains had significantly lowered incidence of colon tumor and colon tumor multiplicity. Cyclooxygenase-2 enzyme activity (the enzyme implicated in colon tumor development) was significantly lower in the colon tissue of rats fed diets containing milk fermented with 4 EPS-producing and 1 non-producing cultures than that in rats fed diets supplemented with GDL-acidified milk. Different EPS-positive cultures produced fermented milks with distinct rheological characteristics and levels of ropiness. No relationship was found between rheological properties or level of ropiness of fermented milk and its chemopreventive effect.     

Volatile organic compounds profile during milk fermentation by Lactobacillus pentosus and correlations between volatiles flavor and carbohydrate metabolism

Flavor, as one of the most important properties determining the acceptability and preference of fermented milks, is influenced by compositional and processing factors. In this study, we focused on the volatile organic compounds related to flavor during milk fermentation by Lactobacillus pentosus according to electronic nose analysis. Xylose (1% addition) metabolized by Lb. pentosus strongly affects the flavor of yogurt, with the potent volatile organic compounds of ethanol (3.08%), 2,3-butanedione (7.77%), and acetic acid (22.70%) detected using solid-phase microextraction coupled with gas chromatography-mass spectrometry analysis. Sensoryanalysis also showed skimmed yogurt fermented by Lb. pentosus with 1% xylose had the unique scores of sourness (acetic acid) and butter flavor (2,3-butanedione). Furthermore, α-acetolactate synthase and α-acetolactate decarboxylase in carbohydrate metabolism play important roles in milk fermentation. Under preferable conditions (pH 5.5, 42°C) for α-acetolactate synthase and α-acetolactate decarboxylase, the relative content of potent flavor compound 2,3-butanedione was 10.13%, which was 2.55% higher than common culture condition (pH 4.5, 37°C), revealing that xylose metabolized by Lb. pentosus has potential values for the milk product industry, such as the acceptability and preference of fermented milk product.     

鱼油风味酸奶的开发及稳定性评价

以全脂奶粉、鱼油、白砂糖为主要原料,以保加利亚乳杆菌和嗜热链球菌为发酵菌种制作鱼油风味酸奶,进行鱼油风味酸奶配方研究与产品稳定性评价。通过感官评定确定最佳酸奶配方为奶粉添加量10%,鱼油添加量0.15%,白砂糖添加量8%。核磁共振与显微观察结果表明,鱼油添加后未对酸奶水分分布产生影响,鱼油在酸奶中呈均匀分布。电子鼻分析结果表明,与原味酸奶相比,鱼油风味酸奶风味更为融合,更接近原味商业酸奶。以储存期pH、酸度、流变特性、感官评价为指标对鱼油风味酸奶进行了稳定性评价,结果表明鱼油风味酸奶在储存期内可保持良好的稳定性,鱼油的添加可有效避免酸奶的过度酸化,使酸奶保持良好的口感。     

不同胞外多糖产生特性的乳酸菌菌种对酸乳品质的影响

不同EPS产生量的两种乳酸菌，用于酸乳的生产，在4种不同菌株组合形成的发酵乳中，球、杆菌比例和乳糖消耗基本一致，但是在粘度、EPS产生、EPS的单糖组成、对乳清析出的影响，以及感官等方面，均有区别。实验结果表明，在酸乳生产中，以产EPS的德氏乳杆菌保加利亚亚种不产EPS的唾液链球菌嗜热亚种组合较好。     

Type of starter culture influences on structural and sensorial properties of low protein fermented gels

Organoleptic properties of skimmed milk fermented gels are progressively demanding to produce optimal quality yogurts. Chr‐Hansen trademark registered cultures were used to produce low‐protein (3.4%) gels to assess the ability to redesign the sensorial and textural properties with the choice of starter culture. Resulting gels were assessed for sensorial, textural, rheological, and microstructural properties and compared with a commercial control (4.5% protein). Mouth thickness, syneresis, firmness, elasticity, and consistency values were lower for polysaccharides‐producing cultures. Such cultures contributed to the higher creaminess and tended to give higher ropiness. Observed differences among microstructures of the gel were minute. Microstructural and rheological data corresponded and reflected the instrumental and sensory interpretations. Strong correlations were observed between sensory and instrumental data. Nonprobiotics cultures resulted in promising overall gel properties compared with probiotic cultures according to the principal component analysis. Yet probiotic cultures resulted in lower syneresis than nonprobiotic cultures. Thus, the choice of bacterial culture modifies the sensorial and textural properties of fermented gel with strong correlations, as a result of altered gel network formation with the production of polysaccharides. Inferior textural and sensorial quality aspects, particularly at low protein levels, have negative impact on consumer demand of low protein yogurts. Thus, we attempted to gain required gel textural and sensorial properties with a choice of starter culture with a low protein level. Resulting gel properties at lowered protein content with different starter cultures are not fully known. The present study compares the effect of probiotic and nonprobiotic starter cultures on gel properties, as gel texture and sensory properties are of great interest and thus not willing to be compromised. In addition, we examined the overall texture profile of studied cultures and correlate with sensory properties. Therefore, reducing protein level in milk and achieving required gel properties with the choice of appropriate starter culture is of great commercial interest as a cost‐cutting strategy to produce low‐cost optimum quality yogurt.     

柠檬红茶酸奶的加工工艺研究

以柠檬、红茶和鲜奶为主要原料,以嗜热性链球菌和保加利亚乳杆菌为发酵剂,经发酵制成一种新型固体酸奶。通过正交试验方法和感官评定,确定该产品的最佳配方和生产工艺为：柠檬红茶汁5%,蔗糖和蜂蜜4%,保加利亚乳杆菌和嗜热性链球菌3％,发酵温度42℃、发酵时间5h。     

Technological properties of milks fermented with thermophilic lactic acid bacteria at suboptimal temperature

In the present work the synergistic relationship between different strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus was studied at optimal (44 degrees C) and suboptimal temperatures (30 degrees C). Acidification, viscosity, whey syneresis, and bacterial concentration of the final product were evaluated on single-strain and mixed cultures after 24 h at 30 degrees C and 6 h at 44 degrees C. Three pairs of strains (LBB + CP2, LBP + CP2, and LBR + CP2) showed synergistic effect, which was reflected by the viscosity and syneresis of the coagulum. These results were more significant when cultures were incubated at 30 degrees C, reaching apparent viscosity values of 19 to 28 mPa x s. On the other hand, lactobacilli cultures enhanced the growth of two streptococci strains (CP2 and CP4). These results were confirmed by cultures of streptococci supplemented with supernatants of culture of lactobacilli. Those supernatants stimulate the viscosity produced by CP2 and CP4 strains and reduce the syneresis of all cultures of streptococci. Neither the increase of viscosity nor reduction of syneresis could be attributed to a decrease of pH.     

Amylose–Potassium Oleate Inclusion Complex in Plain Set‐Style Yogurt

Health and wellness aspirations of U.S. consumers continue to drive the demand for lower fat from inherently beneficial foods such as yogurt. Removing fat from yogurt negatively affects the gel strength, texture, syneresis, and storage of yogurt. Amylose–potassium oleate inclusion complexes (AIC) were used to replace skim milk solids to improve the quality of nonfat yogurt. The effect of AIC on fermentation of yogurt mix and strength of yogurt gel was studied and compared to full‐fat samples. Texture, storage modulus, and syneresis of yogurt were observed over 4 weeks of storage at 4 °C. Yogurt mixes having the skim milk solids partially replaced by AIC fermented at a similar rate as yogurt samples with no milk solids replaced and full‐fat milk. Initial viscosity was higher for yogurt mixes with AIC. The presence of 3% AIC strengthened the yogurt gel as indicated by texture and rheology measurements. Yogurt samples with 3% AIC maintained the gel strength during storage and resulted in low syneresis after storage for 4 wk.     

Technological characterization of lactococci isolated from traditional Chinese fermented milks

To screen lactic acid bacteria for starter cultures in cheese production, 21 Lactococcus strains previously isolated from natural fermented milk and koumiss made by herdsman families in the Xinjiang, Gansu, and Qinghai provinces of China were evaluated for optimal growth temperature, acidification activity, proteolytic activity, aminopeptidase activity, and autolytic activity. All isolates presented low acidification rates, and the pH value did not reach 5.3 after 6 h of inoculation in sterile reconstituted skim milk at 30°C. Strains X9C2 and T7C showed the highest proteolytic activity of 24.67 and 23.58 mg of glycine/L of milk, respectively. For aminopeptidase activity, strains X9C2 and T1C2 displayed the highest activities of 30.56 and 27.70 U/mg of protein using l-leucine-p-nitroanilide as substrate, respectively. Autolytic activity in simulated cheese-like buffer ranged from 7.45 to 34.76%, and strains Q14C2 and Q16C showed the highest values of 34.76 and 34.20%, respectively. Collectively, one main finding is that some technological characteristics of Lactococcus isolates from Chinese traditional fermented products varied greatly. Some isolates with potentially important properties could be valuable for application as starter cultures of cheese or could constitute a mixed culture.     

Contribution of Streptococcus thermophilus to growth-stimulating effect of yogurt on rats

The origin of the growth-stimulating factor in yogurt was studied in rats fed liquid or freeze-dried diets of milk, yogurt, milks fermented individually by Streptococcus thermophilus and Lactobacillus bulgaricus, milks to which cells of Streptococcus thermophilus and Lactobacillus bulgaricus were added. Diets containing sonicated cells, cell supernatant, and cell fractions also were fed. Milk fermented by Streptococcus thermophilus and milk plus Streptococcus thermophilus cells stimulated growth as effectively as did yogurt. That finding and the absence of stimulation in rats fed Lactobacillus bulgaricus showed that Streptococcus thermophilus is responsible for stimulation of growth by yogurt. Growth was stimulated by an intracellular factor and not by fermentative changes in the milk.     

酸乳后酸化影响因子的初步研究

酸乳在贮存、运输、销售、食用前这一过程中发生后酸化现象,导致酸乳无法长期贮存,酸乳贮存期短这一缺陷一直困扰着酸乳的消费。文中从酸乳发酵时保加利亚乳杆菌与嗜热链球菌比例的改变、弱后酸化发酵剂的筛选和酸乳发酵后添加乳酸链球菌素等方面探讨了控制酸乳后酸化的措施。通过改变嗜热链球菌与保加利亚乳杆菌的比例、酸乳发酵后添加乳酸链球菌素对控制酸乳后酸化有一定的效果,但成本较高。控制酸乳后酸化的最好方法是选育在高温条件下产酸较强,在低温条件下产酸甚微的菌株作为发酵剂,并发现采用汉森公司的YF-L812发酵的酸乳后酸化程度很小,在6℃条件下贮存30 d时酸度仅上升了14.5°T。     

Effect of Inoculation Level of Lactobacillus rhamnosus and Yogurt Cultures on Conjugated Linoleic Acid Content and Quality Attributes of Fermented Milk Products

ABSTRACT:  The effect of inoculation concentration of Lactobacillus rhamnosus and yogurt cultures and storage time on conjugated linoleic acid (CLA) content and quality attributes of fermented milk products was determined. Yogurt culture ( Lactobacillus bulgaricus and Streptococcus thermophilus, 1:1 ratio, YC ) , L. rhamnosus (LB), and LB co-cultured with yogurt culture, were inoculated at 10⁶, 10⁷, 10⁸ CFU/mL into a milk with hydrolyzed soy oil as the lipid source. CLA content, microbial counts, acidity, texture, and volatile flavor profile of the fermented milk products were stable during storage at 4 °C for 14 d. Total CLA contents ranged from 0.51 to 1.00 mg CLA/g lipid following 14 d of storage. Inoculation level of L. rhamnosus and yogurt cultures had no significant effect on CLA content and texture, but affected acidity and the volatile flavor profile of the fermented milk products. The fermented milk products produced by L. rhamnosus co-cultured with yogurt culture with 10⁷ CFU/mL total inoculation level resulted in a high CLA content and desirable quality characteristics. This research demonstrated that the optimal inoculation concentration and the combination of L. rhamnosus and yogurt cultures were important factors to produce fermented milk products with CLA content and acceptable quality attributes.