Fermentative properties of starter culture during manufacture of kefir with new prebiotics derived from lactulose

The fermentation properties of a starter culture during kefir manufacture was studied with the inclusion of one emerging lactulose-derived oligosaccharide prebiotic and with the well-recognised galactooligosaccharide and lactulose prebiotics at different doses (2 and 4%). Microbial growth, glycerol, lactic and citric acids and short-chain fatty acids and carbohydrate utilisation during fermentation and cold storage of control and prebiotic supplemented kefirs were determined. Prebiotic levels remained unaltered during fermentation (24 h) and storage (28 days), with the exception of a decrease (7.3%) of lactulose in kefir with 4% prebiotic. Consequently, the viability of lactic acid bacteria and yeasts, as well as the pH or level of fermentation metabolites was similar for all kefirs. Therefore, our data highlight the suitability of kefir as a matrix for the consumption of a variety of prebiotics, including that of novel synthesis as lactulose-derived oligosaccharides, widening their potential food uses and applications.     

Impact of the synbiotic combination of Lactobacillus casei shirota and oligofructose-enriched inulin on the fecal volatile metabolite profile in healthy subjects

Scope: Hypothesis‐driven approaches have mainly focused on the quantification of SCFAs as mediators of beneficial effects of synbiotics. However, the emergence of metabolite profiling strategies allows to evaluate the colonic metabolism from a top‐down approach. In the present study, we evaluated the impact of a synbiotic combination on fecal metabolite profiles. Methods and results: A synbiotic combination (Lactobacillus casei Shirota cells+oligofructose‐enriched inulin) was evaluated in nine healthy volunteers. Before the start, during and after 4‐wk treatment, fecal samples were obtained. GC‐MS technology was applied to analyze the volatile metabolites. Application of a Type III test revealed that the metabolite profiles from the three conditions were significantly different. We identified three volatile organic compounds, acetate, dimethyl trisulfide and ethyl benzene, which were significantly affected. The acetate levels increased, whereas the dimethyl trisulfide levels decreased during and after the intervention. For ethyl benzene only an effect during the synbiotic intervention period was observed. Conclusion: We report a detailed analysis of the influence of L. casei Shirota combined with oligofructose‐enriched inulin on fermentation metabolites. Our results indicated a stimulation of saccharolytic fermentation and, importantly, a reduction of potentially toxic protein fermentation metabolites dimethyl trisulfide and ethyl benzene attended these effects.     

Metabolite profiling of doenjang,fermented soybean paste,during fermentation

BACKGROUND: A fermented soybean paste known as doenjang is a traditional fermented food that is widely consumed in Korea. The quality of doenjang varies considerably by its basic ingredients, species of microflora, and fermentation process. The classification of predefined metabolites (e.g. amino acids, organic acids, sugars and sugar derivatives, and fatty acids) in doenjang samples according to fermentation was performed by using GC‐FID and GC‐MS data sets with the application of a multivariate statistical method. RESULTS: The predominantly produced amino acids included alanine, valine, leucine, isoleucine, proline, glutamine, phenylalanine and lysine, showing remarkable increases in amounts during the later stages of fermentation. Carbonic acid, citric acid, lactic acid and pyrogultamic acid were identified as the major organic acids. Significant amounts of erythrose, xylitol, inositol and mannitol were detected during fermentation. Regarding fatty acids, relatively higher amounts of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid were found in the doenjang at each fermentation time point. Principal component analysis (PCA) successfully demonstrated changes in composition patterns as well as differences in non‐volatile metabolites according to fermentation period. CONCLUSION: A set of metabolites could be determined representing the quality of doenjang during fermentation, and which might also be correlated with taste ingredients, flavour, nutrition, and physiology activities that are claimed to be dependent on the quality control of commercial doenjang. Copyright © 2010 Society of Chemical Industry     

Yeast and bacterial modulation of wine aroma and flavour

Wine is a highly complex mixture of compounds which largely define its appearance, aroma, flavour and mouth‐feel properties. The compounds responsible for those attributes have been derived in turn from three major sources, viz. grapes, microbes and, when used, wood (most commonly, oak). The grape‐derived compounds provide varietal distinction in addition to giving wine its basic structure. Thus, the floral monoterpenes largely define Muscat‐related wines and the fruity volatile thiols define Sauvignon‐related wines; the grape acids and tannins, together with alcohol, contribute the palate and mouth‐feel properties. Yeast fermentation of sugars not only produces ethanol and carbon dioxide but a range of minor but sensorially important volatile metabolites which gives wine its vinous character. These volatile metabolites, which comprise esters, higher alcohols, carbonyls, volatile fatty acids and sulfur compounds, are derived from sugar and amino acid metabolism. The malolactic fermentation, when needed, not only provides deacidification, but can enhance the flavour profile. The aroma and flavour profile of wine is the result of an almost infinite number of variations in production, whether in the vineyard or the winery. In addition to the obvious, such as the grapes selected, the winemaker employs a variety of techniques and tools to produce wines with specific flavour profiles. One of these tools is the choice of microorganism to conduct fermentation. During alcoholic fermentation, the wine yeast Saccharomyces cerevisiae brings forth the major changes between grape must and wine: modifying aroma, flavour, mouth‐feel, colour and chemical complexity. The wine bacterium Oenococcus oeni adds its contribution to wines that undergo malolactic fermentation. Thus flavour‐active yeasts and bacterial strains can produce desirable sensory results by helping to extract compounds from the solids in grape must, by modifying grape‐derived molecules and by producing flavour‐active metabolites. This article reviews some of the most important flavour compounds found in wine, and their microbiological origin.     

紫红曲霉与酿酒酵母共发酵对红曲甜米酒风味的影响

研究紫红曲霉和酿酒酵母在红曲甜米酒发酵过程中风味物质形成的作用。利用紫红曲霉和酿酒酵母以不同比例复配进行甜米酒发酵,监测微生物、有机酸和挥发性香气成分的动态变化。结果表明,以发酵12 h为分界点,酵母菌先增加后减少,而红曲菌则同步呈先减少后增加的趋势;添加红曲菌能够显著促进酵母菌增殖。发酵过程中,酒样的糖度不断降低,酒精度不断增加,发酵36 h后有机酸(乳酸、乙酸、柠檬酸、酒石酸)含量迅速增加;随着红曲菌添加量的提高,糖度减少,酒精和有机酸含量均显著增加。发酵过程共检出47种挥发性组分,包括6种醇类、14种酸类、12种酯类、8种醚类、4种醛酮类、2种醌类和1种酚类,其中高级醇类(苯乙醇、香草醇、异丁醇)和酯类(对羟基扁桃酸乙酯)相对含量较高;酸类含量随发酵时间的延长不断减少,而醇类、酯类和醚类含量则不断增加;添加红曲菌使酯类在发酵前期迅速增加,显著提高其含量。综上所述,添加红曲菌共发酵,促进了酿酒酵母增殖,显著增加了有机酸和酯类等挥发性组分的种类和含量,对甜米酒风味物质的丰富具有重要贡献。     

Effects of 3-nitrooxypropanol on methane emission,digestion, and energy and nitrogen balance of lactating dairy cows

The objective was to measure effects of 3-nitrooxypropanol (3NP) on methane production of lactating dairy cows and any associated changes in digestion and energy and N metabolism. Six Holstein-Friesian dairy cows in mid-lactation were fed twice daily a total mixed ration with maize silage as the primary forage source. Cows received 1 of 3 treatments using an experimental design based on two 3 × 3 Latin squares with 5-wk periods. Treatments were a control placebo or 500 or 2,500 mg/d of 3NP delivered directly into the rumen, via the rumen fistula, in equal doses before each feeding. Measurements of methane production and energy and N balance were obtained during wk 5 of each period using respiration calorimeters and digestion trials. Measurements of rumen pH (48 h) and postprandial volatile fatty acid and ammonia concentrations were made at the end of wk 4. Daily methane production was reduced by 3NP, but the effects were not dose dependent (reductions of 6.6 and 9.8% for 500 and 2,500 mg/d, respectively). Dosing 3NP had a transitory inhibitory effect on methane production, which may have been due to the product leaving the rumen in liquid outflow or through absorption or metabolism. Changes in rumen concentrations of volatile fatty acids indicated that the pattern of rumen fermentation was affected by both doses of the product, with a decrease in acetate:propionate ratio observed, but that acetate production was inhibited by the higher dose. Dry matter, organic matter, acid detergent fiber, N, and energy digestibility were reduced at the higher dose of the product. The decrease in digestible energy supply was not completely countered by the decrease in methane excretion such that metabolizable energy supply, metabolizable energy concentration of the diet, and net energy balance (milk plus tissue energy) were reduced by the highest dose of 3NP. Similarly, the decrease in N digestibility at the higher dose of the product was associated with a decrease in body N balance that was not observed for the lower dose. Milk yield and milk fat concentration and fatty acid composition were not affected but milk protein concentration was greater for the higher dose of 3NP. Twice-daily rumen dosing of 3NP reduced methane production by lactating dairy cows, but the dose of 2,500 mg/d reduced rumen acetate concentration, diet digestibility, and energy supply. Further research is warranted to determine the optimal dose and delivery method of the product.     

Chemical composition of naturally fermented buttermilk

Naturally fermented buttermilk, prepared from soured cream or milk, was collected during two seasons from sixteen farms in northern Ethiopia, to study chemical composition and flavour compounds. Protein, fat, organic acids, carbohydrates and volatile compounds were quantified using Kjeldahl, Gerber, high‐pressure liquid chromatography (HPLC) and headspace GC methods, respectively. Widely differing concentrations of organic acids and volatile compounds among samples indicated variable fermentation in the products. This indicates the need for the introduction of the standardisation of the process to supply the market with homogenous buttermilk products.     

脂肪和蛋白质含量对稀奶油-乳清发酵液的影响

本文研究了不同脂肪含量(7%、14%和21%)和蛋白质含量(2.3%和4.6%)对稀奶油-乳清发酵液的影响,研究新型风味发酵体系组成和发酵特性之间的关系。测定在发酵过程中活菌数、酸度和p H的变化,利用顶空-固相微萃取(HS-SPME)和气相色谱-质谱联用(GC-MS)分析发酵60 h后发酵液中挥发性风味物质,并对其进行感官评定。结果发现:蛋白质含量较低时,脂肪含量增加对乳酸菌增殖和p H值无影响,蛋白质含量较高时,脂肪含量增加抑制乳酸菌增殖和p H值下降;脂肪含量增加促进产香和挥发性风味物质增加,抑制菌种产酸。蛋白质含量增加促进乳酸菌增殖和挥发性风味物质明显增加,使p H值下降减缓;脂肪含量较低时,蛋白质含量增加促进产酸,脂肪含量较高时,抑制产酸。发酵60 h后,体系中主要挥发性风味物质有乙酸、丁酸、2,3-丁二酮和3-羟基-2-丁酮。通过感官评定,14%脂肪和4.6%蛋白质的发酵液具有最佳的组织状态以及最强的特征性酸奶风味。     

Effect of Fermentation Temperature on the Volatile Composition of Kimchi

This study was conducted to evaluate the effect of fermentation temperature on the volatile composition in Kimchi. Kimchi was fermented at 2 temperature conditions (4 and 20 °C). Volatile compounds of Kimchi samples were analyzed during the fermentation periods using the dynamic headspace gas chromatography‐mass spectrometry method. The optimum ripening time for the Kimchi fermented at 4 °C was 35 d, and that of 20 °C was 2 d. The pH at the optimum ripening time was 4.97 and 4.41, and the titratable acidity was 0.59% and 0.76% for the Kimchi fermented at 4 and 20 °C, respectively. Forty different types of volatile compounds, including alcohol, aldehyde, ester, and sulfur compounds, were identified. The Kimchi fermented at 20 °C produced greater amounts of volatile compounds than that at 4 °C. The amounts of most volatiles increased as the fermentation time increased, but those of aldehydes decreased rapidly during both 4 and 20 °C fermentation. Organic acids, ester, and nitriles were detected only in Kimchi fermented at 20 °C. The amounts of dimethyl disulfide, methyl‐2‐propenyl disulfide, and di‐2‐propenyl disulfide produced from the Kimchi fermented at 20 °C were more than 2‐times of those at 4 °C. Therefore, it is concluded that the strong pungent odor of Kimchi fermented at 20° C is probably due to the high amount of organic acids (low pH) and sulfur compounds (dimethyl disulfide, methyl‐2‐propenyl disulfide, and di‐2‐propenyl disulfide) between the 2 Kimchi.     

橡木片对霞多丽葡萄酒发酵的影响

该文研究在发酵过程中添加法国中度烘烤橡木制品对霞多丽干白葡萄酒的影响,增加霞多丽干白葡萄酒酿造工艺的选择性.接种酵母菌进行酒精发酵的同时在葡萄汁中添加法国中度烘烤橡木制品,未添加橡木制品发酵的样品为对照样,分别在发酵降糖50％和酒精发酵结束时测定理化指标、有机酸与挥发性化合物的含量.结果表明:添加橡木制品发酵对霞多丽葡萄酒化学物质有一定影响,其总高级醇、总橡木挥发性化合物含量较高.而对照样的葡萄酒中总酸和总酯含量较高,总高级醇含量是添加橡木制品发酵的葡萄酒中总高级醇含量的70％～85％,而酸类物质含量是添加橡木制品发酵葡萄酒的2倍.     

Bioactive compounds and antioxidant activities of the Korean lotus leaf (Nelumbo nucifera) condiment: volatile and nonvolatile metabolite profiling during fermentation

Recent studies have shown that lotus (Nelumbo nucifera) leaf has various pharmacological effects. However, there have been no scientific investigations into these leaves as a fermented crude liquid. This study compared the in vitro antioxidant capacity of lotus leaf‐fermented broth (LLFB) with that of 57° Brix sugar broth (SB) as a control over a period of 6 months based on 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging activity, ferric ion reducing power, superoxide dismutase‐like activity, tyrosinase inhibition and nitrite scavenging activity. The dominant species during fermentation process were Leuconostoc lactis and Saccharomyces cerevisiae for LLFB. A total of thirty‐six metabolites such as alkaloids, fatty acids, organic acids, phenolics, sugar and sugar derivatives, ethyl esters and monoterpenoids differed between SB and LLFB using gas chromatography–mass spectrometry. Meanwhile, nine volatile compounds in LLFB contributed pleasant, slightly sweetish and fruity odour of condiment and sensory evaluation score of 4.06 ± 1.52 in the proportion of 1:9 LLFB/water.     

Changes in volatile compounds and related biochemical profile during controlled fermentation of cv. Conservolea green olives

The effect of controlled fermentation processes on the profile of volatile and other biochemical compounds of cv. Conservolea green olives processed by the Spanish method was studied. The different treatments included: (a) inoculation with a commercial starter culture of Lactobacillus pentosus, (b) inoculation with a wild strain of Lactobacillus plantarum isolated from a previous fermentation, (c) uninoculated spontaneous process (control). Microbial growth, pH, titratable acidity, reducing sugars, organic acids and volatile compounds were monitored. Starter cultures were effective in establishing an accelerated fermentation process. Both were able to reduce the survival period of Enterobacteria by 7 days, minimizing thus the likelihood of spoilage. Higher acidification of the brines and faster pH drop was observed in inoculated processes, with L. pentosus presenting better performance than the wild strain of L. plantarum. Lactic and acetic were the major organic acids detected by HPLC, the concentration of which increased in the course of fermentation. Citric and malic acids were also present in the brines but they were degraded completely within the first 2 weeks of fermentation. Ethanol, methanol, acetaldehyde, ethyl acetate, isobutyric acid were the major volatile compounds identified by GC. Their concentration varied greatly among the fermentation processes, reflecting varying degrees of microbial activity in the brines.     

Effects of 4 Probiotic Strains in Coculture with Traditional Starters on the Flavor Profile of Yogurt

To study the influence of probiotics on the flavor profile of yogurt, 4 probiotics, including Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus casei, were cofermented with traditional starters. The changes of bacterial growth, acid contents and volatile compounds of yogurt were investigated during fermentation and refrigerated storage. The strains that exhibited a low growth rate in milk did not significantly affect the bacterial population dynamics, acidity, or organic acid content during fermentation and storage. However, high viability and enhancement of postacidification were clearly observed in the samples that contained strains with a high growth rate in milk, particularly L. casei. A total of 45 volatile compounds, detected in most samples, were identified by headspace solid‐phase micro‐extraction followed by gas chromatography‐mass spectrometry. Among these compounds, ketones and aldehydes were the most abundant. The presence of either L. rhamnosus or L. plantarum did not significantly affect the major volatile compounds, while contributions of L. casei and L. acidophilus were found in the formation of minor volatile metabolites. Electronic nose measurements exhibited a good discrimination of samples that contained different probiotics during refrigerated storage.     

Volatile Compounds Produced by Lactobacillus paracasei During Oat Fermentation

This study investigated the profiles of volatile compounds produced by Lactobacillus paracasei during oat fermentation using gas chromatography‐mass spectrometry coupled with headspace solid‐phase microextraction method. A total of 60 compounds, including acids, alcohols, aldehydes, esters, furan derivatives, hydrocarbons, ketones, sulfur‐containing compounds, terpenes, and other compounds, were identified in fermented oat. Lipid oxidation products such as 2‐pentylfuran, 1‐octen‐3‐ol, hexanal, and nonanal were found to be the main contributors to oat samples fermented by L. paracasei with the level of 2‐pentylfuran being the highest. In addition, the contents of ketones, alcohols, acids, and furan derivatives in the oat samples consistently increased with the fermentation time. On the other hand, the contents of degradation products of amino acids, such as 3‐methylbutanal, benzaldehyde, acetophenone, dimethyl sulfide, and dimethyl disulfide, decreased in oat samples during fermentation. Principal component analysis (PCA) was applied to discriminate the fermented oat samples according to different fermentation times. The fermented oats were clearly differentiated on PCA plots. The initial fermentation stage was mainly affected by aldehydes, whereas the later samples of fermented oats were strongly associated with acids, alcohols, furan derivatives, and ketones. The application of PCA to data of the volatile profiles revealed that the oat samples fermented by L. paracasei could be distinguished according to fermentation time.     

Chromohalobacter is a Causing Agent for the Production of Organic Acids and Putrescine during Fermentation of Ganjang,a Korean Traditional Soy Sauce

Ganjang, a Korean traditional fermented soy sauce, is prepared by soaking doenjang‐meju (fermented soybeans) in approximately 20% (w/v) solar salt solution. The metabolites and bacterial communities during ganjang fermentation were simultaneously investigated to gain a better understanding of the roles of the microbial population. The bacterial community analysis based on denaturing gradient gel electrophoresis of 16S rRNA gene sequences showed that initially, the genus Cobetia was predominant (0 to 10 d), followed by Bacillus (5 to 74 d), and eventually, Chromohalobacter became predominant until the end of the fermentation process (74 to 374 d). Metabolite analysis using ¹H‐NMR showed that carbon compounds, such as fructose, galactose, glucose, and glycerol, probably released from doenjang‐meju, increased rapidly during the early fermentation period (0 to 42 d). After removal of doenjang‐meju from the ganjang solution (42 d), the initial carbon compounds remained nearly constant without the increase of fermentation products. At this point, Bacillus species, which probably originated from doenjang‐meju, were predominant, suggesting that Bacillus is not mainly responsible for ganjang fermentation. Fermentation products including acetate, lactate, α‐aminobutyrate, γ‐aminobutyrate, and putrescine increased quickly with the rapid decrease of the initial carbon compounds, while Chromohalobacter, probably derived from the solar salts, was predominant. Multivariate redundancy analysis indicated that the Chromohalobacter population was closely correlated with the production of the organic acids and putrescine during the ganjang fermentation. These results may suggest that Chromohalobacter is a causing agent responsible for the production of organic acids and putrescine during ganjang fermentation and that the solar salts, not doenjang‐meju, is an important microbial source for ganjang fermentation.     

Changes in Volatile Compounds of Chinese Rice Wine Wheat Qu During Fermentation and Storage

In order to understand the optimum time of using wheat Qu for Chinese rice wine brewing on the basis of the contents of volatile compounds, changes in volatile compounds of Chinese rice wine wheat Qu during fermentation and storage were studied by headspace solid phase microextraction (HS‐SPME) combined with gas chromatography‐mass spectrum (GC‐MS). It was shown that the concentrations of most groups of volatiles, including alcohols, aldehydes, aromatic compounds, phenols, sulphides, furans, and nitrogen‐containing compounds, increased progressively from the first day to the fourth day of fermentation, and then decreased gradually. A minority group of volatile compounds such as acids and ketones peaked after one day of fermentation, esters increased slowly from the first day to the eighth day of fermentation. Lactones and esters rose slowly from the first day to the fifteenth day of fermentation respectively, and decreased afterwards, whereas the evolution of terpenes was somewhat erratic. These results suggested that changes in the concentrations of most groups of volatile compounds during fermentation and storage were uniform. This made it possible to determine the optimum level of fermentation and storage for Chinese rice wine wheat Qu on the basis of the contents of volatile compounds.     

Effects of Different Salt Treatments on the Fermentation Metabolites and Bacterial Profiles of Kimchi

The effects of purified salt (PS) and mineral‐rich sea salt (MRS), both with different mineral profiles, on kimchi fermentation were studied using a culture‐dependent 16S rRNA sequencing technique and mass‐based metabolomic analysis. The different mineral profiles in the fermentation medium caused changes in the bacterial profiles of the 2 kimchi products. An increase of Leuconostoc species in MRS‐kimchi decreased the Lactobacillus/Leuconostoc ratio, which led to changes in metabolites (including sugars, amino acids, organic acids, lipids, sulfur compounds, and terpenoids) associated with kimchi quality. Although further studies on the relationship between these salt types and kimchi fermentation are needed, these results suggested that the MRS treatment had positively affected the changes of the kimchi mineral contents, bacterial growth, and metabolite profiles, which are linked to kimchi quality.     

Evolution of volatile compounds in papaya wine fermented with three Williopsis saturnus yeasts

The changes in volatile compounds during papaya juice fermentation with three Williopsis saturnus yeasts were investigated in this study. Time‐course papaya juice fermentations were carried out using three Williopsis saturnus yeasts: W. saturnus var. mrakii NCYC2251, W. saturnus var. saturnus NCYC22 and W. saturnus var. sargentensis NCYC2727. Changes in yeast cell population, Brix and pH were similar among the three yeasts, which preferentially utilised glucose over fructose while partially degrading l ‐malic acid. A range of volatile compounds were produced during fermentation including fatty acids, alcohols and esters with esters being the most abundant volatile compounds produced. Benzyl isothiocyanate, butyric acid, 2‐ethylhexanol, benzaldehyde and β‐damascenone present in the papaya juice were metabolised to trace levels during fermentation. There were significant variations among the three yeasts in their ability to produce and metabolise volatile compounds during fermentation. The study suggests that papaya juice fermentation with W. saturnus yeasts is able to result in the formation of a more complex aroma compounds.     

葡萄酵素发酵过程中代谢产物的动态变化

为探究葡萄酵素接种发酵过程中物质变化规律,以葡萄为原料,分析了接种酵母菌、醋酸菌以及乳酸菌发酵过程中总酸、有机酸、乙醇体积分数、总酚、主要功效酶（超氧化物歧化酶及β-葡萄糖苷酶）以及香气成分等代谢产物的变化。结果表明：发酵过程中总酸质量浓度呈先上升后下降的趋势,于第34天达到最大值19.37 g/L;在接种酵母菌（0～6 d）发酵过程中有机酸主要以柠檬酸、苹果酸、酒石酸为主,且乙醇主要产生于该阶段;接种醋酸菌后（6～27 d）主要产生乙酸;而后接种乳酸菌后（27～41 d）,乳酸、柠檬酸质量浓度逐渐增加至最大值102.09 mg/L和67.70 mg/L,而苹果酸减少,此时乙醇体积分数降低至最小值0.47%并趋于稳定;总酚含量与β-葡萄糖苷酶活力的变化趋势一致,主要于接种酵母菌（0～6 d）发酵以及接种乳酸菌发酵后期（34～41 d）产生,超氧化物歧化酶活力总体呈波动上升趋势;葡萄酵素发酵过程中产生的挥发性物质主要以醇类、酸类和酯类化合物为主。以上研究结果可为葡萄酵素的精准制备以及品质优化提供理论依据。