Comparative study of the biochemical changes and volatile compound formations during the production of novel whey-based kefir beverages and traditional milk kefir

Cheese whey (CW) and deproteinised cheese whey (DCW) were investigated for their suitability as novel substrates for the production of kefir-like beverages. Lactose consumption, ethanol production, as well as organic acids and volatile compounds formation, were determined during CW and DCW fermentation by kefir grains and compared with values obtained during the production of traditional milk kefir. The results showed that kefir grains were able to utilise lactose from CW and DCW and produce similar amounts of ethanol (7.8–8.3 g/l), lactic acid (5.0 g/l) and acetic acid (0.7 g/l) to those obtained during milk fermentation. In addition, the concentration of higher alcohols (2-methyl-1-butanol, 3-methyl-1-butanol, 1-hexanol, 2-methyl-1-propanol, and 1-propanol), ester (ethyl acetate) and aldehyde (acetaldehyde) in cheese whey-based kefir and milk kefir beverages were also produced in similar amounts. Cheese whey and deproteinised cheese whey may therefore serve as substrates for the production of kefir-like beverages similar to milk kefir.     

Short communication: Effect of kefir grains on proteolysis of major milk proteins

The effect of kefir grains on the proteolysis of major milk proteins in milk kefir and in a culture of kefir grains in pasteurized cheese whey was followed by reverse phase-HPLC analysis. The reduction of κ-, α-, and β-caseins (CN), α-lactalbumin (α-LA), and β-lactoglobulin (β-LG) contents during 48 and 90 h of incubation of pasteurized milk (100 mL) and respective cheese whey with kefir grains (6 and 12 g) at 20°C was monitored. Significant proteolysis of α-LA and κ-, α-, and β-caseins was observed. The effect of kefir amount (6 and 12 g/100 mL) was significant for α-LA and α- and β-CN. α-Lactalbumin and β-CN were more easily hydrolyzed than α-CN. No significant reduction was observed with respect to β-LG concentration for 6 and 12 g of kefir in 100 mL of milk over 48 h, indicating that no significant proteolysis was carried out. Similar results were observed when the experiment was conducted over 90 h. Regarding the cheese whey kefir samples, similar behavior was observed for the proteolysis of α-LA and β-LG: α-LA was hydrolyzed between 60 and 90% after 12 h (for 6 and 12 g of kefir) and no significant β-LG proteolysis occurred. The proteolytic activity of lactic acid bacteria and yeasts in kefir community was evaluated. Kefir milk prepared under normal conditions contained peptides from proteolysis of α-LA and κ-, α-, and β-caseins. Hydrolysis is dependent on the kefir:milk ratio and incubation time. β-Lactoglobulin is not hydrolyzed even when higher hydrolysis time is used. Kefir grains are not appropriate as adjunct cultures to increase β-LG digestibility in whey-based or whey-containing foods.     

Effect of fat replacers on kefir quality

The purpose of the study was to determine the effects of fat replacers on the quality of non‐fat kefir. Skim milk fortified with Dairy Lo^® (DL) and inulin (INU) was fermented with kefir grains to manufacture kefir. The results of compositional, microbiological, rheological and sensorial analyses were compared with whole kefir (WK) and non‐fat kefir (NFK) controls. Results for dry matter, pH and lactic acid ranged between 82.4 and 109.1 g kg^?1, 4.26 and 4.40, and 7.0 and 9.2 g L^?1, respectively. Acetaldehyde and ethanol contents of samples were between 2.89 and 7.28 mg L^?1, and 151.46 and 323.89 mg L^?1, respectively. In all samples, Lactobacillus spp., Streptococcus spp. and yeast counts were between 9.1 and 9.9, 9.3 and 9.9, and 5.2 and 5.6 log cfu mL^?1, respectively. Kefir samples had non‐Newtonian behaviour and pseudoplastic fluid with thixotropy. At the first day, DL had the highest apparent viscosity (3.119 Pa s) while NFK had the lowest value (1.830 Pa s). In the sensory evaluation, odour and taste scores of samples were not different. Dairy Lo^® and inulin could be used without any adverse effect for the production of non‐fat kefir. Copyright © 2009 Society of Chemical Industry     

Kefir Immobilized on Corn Grains as Biocatalyst for Lactic Acid Fermentation and Sourdough Bread Making

Abstract: The natural mixed culture kefir was immobilized on boiled corn grains to produce an efficient biocatalyst for lactic acid fermentation with direct applications in food production, such as sourdough bread making. The immobilized biocatalyst was initially evaluated for its efficiency for lactic acid production by fermentation of cheese whey at various temperatures. The immobilized cells increased the fermentation rate and enhanced lactic acid production compared to free kefir cells. Maximum lactic acid yield (68.8 g/100 g) and lactic acid productivity (12.6 g/L per day) were obtained during fermentation by immobilized cells at 37 °C. The immobilized biocatalyst was then assessed as culture for sourdough bread making. The produced sourdough breads had satisfactory specific loaf volumes and good sensory characteristics. Specifically, bread made by addition of 60% w/w sourdough containing kefir immobilized on corn was more resistant regarding mould spoilage (appearance during the 11^th day), probably due to higher lactic acid produced (2.86 g/Kg of bread) compared to the control samples. The sourdough breads made with the immobilized biocatalyst had aroma profiles similar to that of the control samples as shown by headspace SPME GC‐MS analysis.     

Production of Kefir from Soymilk With or Without Added Glucose, Lactose, or Sucrose

ABSTRACT: The effects of added glucose, lactose, and sucrose on microbial growth, acid, and ethanol production, and galactosidase activity in soymilk fermented with kefir grains were studied. Immediately after the addition of kefir grains to soymilk, the lactic-acid bacterial counts were higher, but the yeast counts were lower than in milk kefir. After fermentation for 32 h, the concentrations of yeast, lactic acid, and ethanol in soymilk were significantly lower than those in milk kefir. Addition of 1% glucose to soymilk stimulated growth of lactic-acid bacteria and yeast, the production of lactic acid and ethanol, and the β-galactosidase activity. Nevertheless α- galactosidase activity was suppressed by 1% glucose.     

Assessment of beverages made from milk,soya milk and whey using Iranian kefir starter culture

Kefir starter culture was used for the production of beverages, and some chemical and microbiological parameter changes were determined during 24 h of fermentation and then after 2, 7, 14, 21 and 28 days of storage at 4 °C. Three different substrates (milk, whey and soya milk) were used as fermentation media for the kefir starter culture. After the fermentation, the carbohydrate content and pH decreased, but the dry matter and fat content of the beverages were not significantly different from their substrates. During storage, lactic acid bacteria in the beverages decreased, while yeasts increased and carbohydrate, ethanol and pH changed significantly in the three beverages.     

Effect of different growth conditions on biomass increase in kefir grains

Kefir is a functional dairy product and the effects of kefir consumption on health have been well documented. Kefir grains have naturally high numbers of lactic acid bacteria and yeasts and are used in manufacturing kefir. The biomass of kefir grains slowly increases after successive fermentations. The effects of adding whey protein isolate, modified whey protein (MWP, fat replacer; Carbery Inc., Cork, Ireland), or inulin to milk and different atmospheric conditions (ambient or 6% CO₂) during fermentation on the increase in biomass of kefir grains were investigated. Reconstituted milks (10% milk powder) enriched with whey protein isolate (2%), MWP (2%), and inulin (2%) were inoculated with kefir grains and fermented in ambient and 6% CO₂ incubators at 25°C until a final pH of 4.6 was reached. Biomass increments of kefir grains were determined weekly over 30 d. Lactic acid bacteria and yeast contents of kefir grains were also determined. The highest biomass increase (392%) was found in kefir grains grown in milk supplemented with whey protein isolate under ambient atmospheric conditions. Application of CO₂ did not provide a significant supporting effect on the biomass of kefir grains. Addition of MWP significantly accelerated the formation of kefir grain biomass (223%). The use of whey protein isolate, MWP, or inulin in milk did not cause any adverse effects on the microbial flora of kefir grains.     

Assessment of hydrolysis of cheese whey and use of hydrolysate for bioproduction of carotenoids by Sporidiobolus salmonicolor CBS 2636

BACKGROUND: The increasing industrial demand for carotenoids has led to growing interest in their bioproduction. The need to reduce production costs has encouraged the use of low‐cost agroindustrial substrates. In this context, this work studied the pretreatment of Mozzarella cheese whey and the use of the pretreated whey as fermentation medium for the bioproduction of carotenoids by Sporidiobolus salmonicolor CBS 2636. RESULTS: Bioproduction was carried out in an orbital shaker using a 10 mL L^?1 inoculum, incubation at 25 °C and a stirring rate of 180 rpm for 120 h in a non‐illuminated environment. The carotenoids were recovered using liquid N₂ combined with dimethyl sulfoxide for cell rupture and an acetone/methanol mixture (7:3 v/v) for extraction. The maximum concentration of total carotenoids obtained was 590.4 µg L^?1 in a medium with 900 g L^?1 cheese whey hydrolysate and 4 g L^?1 K₂HPO₄ at 180 rpm, 25 °C and pH 4. CONCLUSION: The use of enzyme‐hydrolysed cheese whey was more effective in carotenoid bioproduction by S. salmonicolor CBS 2636 than the use of acid‐hydrolysed cheese whey. The concentration of total carotenoids obtained with the enzymatic hydrolysate was six times higher than that obtained with the acid hydrolysate. Copyright © 2009 Society of Chemical Industry     

Inhibitory power of kefir: the role of organic acids

Milk and MRS broth fermented with kefir grains from different households were examined for inhibitory activity toward gram-negative and gram-positive strains. Fermented milk obtained with 10 g per 100 ml of inoculum (final pH 3.32 to 4.25) and MRS broth fermented with 1 and 10 g per 100 ml of inocula (final pH 4.18 to 5.25) had inhibitory power demonstrated by spot test and agar well diffusion assay. This inhibitory effect could be assigned to the undissociated form of lactic and acetic acid produced during the fermentation process. Kefir supernatants inhibited the growth of Escherichia coli 3 in nutrient broth at 37 degrees C for 24 h. However, supernatants of yogurt or milk artificially acidified with lactic and acetic acids allowed the growth of E. coli 3 in the same conditions. A bacteriostatic effect of milk fermented with kefir grains over E. coli 3 was also demonstrated.     

Inferring the role of microorganisms in water kefir fermentations

Water kefir is a slightly alcoholic, lactic and acetic beverage fermented by yeasts, lactic acid bacteria and acetic acid bacteria that are associated with the polysaccharide of the water kefir grains. In this study, the three main metabolic products of microorganisms were evaluated during a traditional 192‐h water kefir fermentation and also after inoculating the microorganisms in fresh medium or sterilised broth from different fermentation stages. The first process to occur was alcoholic fermentation, carried out in particular by Saccharomyces cerevisiae. After 24 h, lactic and acetic acid accumulation was generated by Lactobacillus hilgardii and Acetobacter tropicalis. By the end of fermentation, ethanol had been almost entirely consumed and oxidised to acetic acid, possibly by a dissimilatory route of Acetobacter species. An original hypothetical diagram is proposed for the carbon flux from sucrose, and the metabolic role of the main yeasts and bacteria is assigned for the distinct stages of water kefir fermentation.     

The quality of plain and supplemented kefir from goat's and cow's milk

Goat's or cow's milk was fortified with 2 g/100 g skimmed milk powder (SMP), whey protein concentrate (WPC) or inulin. All the milks were fermented with kefir grains at 25°C for 19 h and stored for 10 days at 5°C. All the kefir samples were analysed on the 1st, 5th and 10th day of storage. The acidity level remained very stable in all the samples during the storage period. Goat's samples have significantly lower viscosity and slightly lower sensory profiles, mostly due to softer consistency. Concentration of ethanol was low, regardless of milk type used, especially for control samples.     

Xanthan gum produced by Xanthomonas campestris from cheese whey: production optimisation and rheological characterisation

BACKGROUND: This aim of this study was the production and rheological characterisation of xanthan gum by Xanthomonas campestris pv. mangiferaeindicae IBSBF 1230 using industrial media and experimental design techniques in a bench bioreactor. RESULTS: The optimised conditions for the production of xanthan starting with 900 mL of cheese whey were 1 g L^?1 magnesium sulphate, 20 g L^?1 potassium phosphate, 28 °C temperature and initial pH 7.2 at 390 rpm agitation and 1.5 vvm aeration, resulting in 36 g L^?1 gum in 72 h. The highest viscosity obtained in the production optimisation study was 1831.34 mPa s at 25 °C with 30 g L^?1 gum. The use of CaCl₂ resulted in the highest solution viscosity under conditions of 25 °C, 1 g L^?1 salt and 46.8 g L^?1 gum, with a value of 1704.53 mPa s. CONCLUSION: In this study, cheese whey, a by‐product of the dairy industry, was used as substrate in the production of xanthan gum, a valuable product in food applications, with optimised high gum production in a bioreactor and a wide range of viscosity values. Copyright © 2009 Society of Chemical Industry     

A review on traditional Turkish fermented non-alcoholic beverages: Microbiota,fermentation process and quality characteristics

Shalgam juice, hardaliye, boza, ayran (yoghurt drink) and kefir are the most known traditional Turkish fermented non-alcoholic beverages. The first three are obtained from vegetables, fruits and cereals, and the last two ones are made of milk. Shalgam juice, hardaliye and ayran are produced by lactic acid fermentation. Their microbiota is mainly composed of lactic acid bacteria (LAB). Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus paracasei subsp. paracasei in shalgam fermentation and L. paracasei subsp. paracasei and Lactobacillus casei subsp. pseudoplantarum in hardaliye fermentation are predominant. Ayran is traditionally prepared by mixing yoghurt with water and salt. Yoghurt starter cultures are used in industrial ayran production. On the other hand, both alcohol and lactic acid fermentation occur in boza and kefir. Boza is prepared by using a mixture of maize, wheat and rice or their flours and water. Generally previously produced boza or sourdough/yoghurt are used as starter culture which is rich in Lactobacillus spp. and yeasts. Kefir is prepared by inoculation of raw milk with kefir grains which consists of different species of yeasts, LAB, acetic acid bacteria in a protein and polysaccharide matrix. The microbiota of boza and kefir is affected from raw materials, the origin and the production methods.     

Improvement of L(+)‐lactic acid production from cassava wastewater by Lactobacillus rhamnosus B 103

BACKGROUND: L (+)‐Lactic acid is used in the pharmaceutical, textile and food industries as well as in the synthesis of biodegradable plastics. The aim of this study was to investigate the effects of different medium components added in cassava wastewater for the production of L (+)‐lactic acid by Lactobacillus rhamnosus B 103. RESULTS: The use of cassava wastewater (50 g L^?1 of reducing sugar) with Tween 80 and corn steep liquor, at concentrations (v/v) of 1.27 mL L^?1 and 65.4 mL L^?1 respectively led to a lactic acid concentration of 41.65 g L^?1 after 48 h of fermentation. The maximum lactic acid concentration produced in the reactor after 36 h of fermentation was 39.00 g L^?1 using the same medium, but the pH was controlled by addition of 10 mol L^?1 NaOH. CONCLUSION: The use of cassava wastewater for cultivation of L. rhamnosus is feasible, with a considerable production of lactic acid. Furthermore, it is an innovative proposal, as no references were found in the scientific literature on the use of this substrate for lactic acid production. Copyright © 2010 Society of Chemical Industry     

Isolation and characterization of the microbial population of different South African kefir grains

Kefir, a slightly acidic fermented milk, is produced by adding lactic acid bacteria and yeasts, in the form of grains, to milk. The bacteria and yeasts present in the kefir grains are known to vary widely. Selective growth media and morphological and biochemical characteristics were used for the isolation and identification of the microbes present in the grains from eight different sources in South Africa. The kefir grains were activated in milk for only 24 h to prevent any changes in the microbial population of the grains. The microbial numbers varied between 6.4 × 10⁴ and 8.5 × 10 ⁸  cfu/g on the media selective for the bacterial species and between 1.5 × 10 ⁵ and 3.7 × 10 ⁸   cfu/g on the media selective for the yeast species. The bacterial genera that were identified included Lactobacillus , Leuconostoc and Lactococcus and the yeast genera included Zygosaccharomyces , Candida and Saccharomyces . The distribution frequencies of the microbes in the different grains were determined and most of the grains were dominated by two microbial species. No pediococci, acetic acid bacteria or propionibacteria were detected.     

Microbiological, physicochemical, and sensory characteristics of kefir during storage

Changes in certain microbiological, physicochemical, and sensory parameters of kefir were studied during refrigerated storage. Kefir batches were prepared using 1% and 5% added kefir grains, and samples for analysis were taken 24 h after inoculation and then after 2, 7, 14, 21, and 28 days of storage at 5 ± 1 °C. After fermentation for 24 h after inoculation, lactobacilli and lactococci were present at levels of 10⁸ cfu/ml, and yeasts and acetic acid bacteria were present at levels of 10⁵ and 10⁶ cfu/ml, respectively. The lactic acid flora decreased by about 1.5 log units between days 7 and 14 and then stabilized at that level. Yeast and acetic acid bacterial counts, lactose, and pH all remained constant over the storage period, while the total fat content and dry matter decreased. The percentage inoculate did exert an influence, and the sample batches made using 1% added kefir grains had higher lactic acid bacterial counts, lactose, and pH, while the sample batches made using 5% added kefir grains had higher yeast and acetic acid bacterial counts and viscosity. The total fat and dry matter contents were similar in both sample batches. Sensory analysis of the kefir samples revealed maximum acceptability levels in the first 2 days of storage.     

Variation in short chain fatty acid and ethanol concentration resulting from the natural fermentation of wheat and barley for inclusion in liquid diets for pigs

Fifty‐six samples of wheat and 44 samples of barley were taken, at harvest, from locations across the UK. Lactic acid bacteria (LAB) and yeasts were enumerated before the samples were ground. Following grinding, triplicate 30‐g samples of each cereal were mixed with sterile distilled water and incubated at 30, 35 or 40 °C. Samples were taken immediately after mixing and at 24‐h intervals for analysis of short‐chain fatty acids (SCFA) and ethanol by isocratic ion‐exclusion liquid chromatography. The number of LAB and yeasts present in samples ranged from 0 to 5.0 (mean 2.25 ± 1.31) and 3.30 to 6.25 (mean 4.96 ± 0.74) log₁₀ colony‐forming units (cfu) ml^?1 respectively. At 30 °C the mean concentrations (mmol l^?1) of SCFAs and ethanol were, lactic acid 59.6 ± 40.0 (range 0.14–134.9), acetic acid 23.2 ± 11.1 (range 2.9–51.4), butyric acid 17.2 ± 16.8 (range 0.0–62.2) and ethanol 15.0 ± 9.0 (range 4.6–53.7) respectively. After fermentation for 24 h only 9 of 300 fermentations produced more than 75 mmol l^?1 lactic acid, which has previously been demonstrated to prevent the growth of Salmonella in liquid pig feed. Fermenting at 35 or 40 °C had no effect on lactic acid concentration but significantly (p < 0.001) increased the concentrations of acetic and butyric acids and ethanol. These results indicate that natural fermentation cannot be relied upon to produce levels of SCFAs that will prevent the proliferation of enteropathogens. Copyright © 2004 Society of Chemical Industry     

Determination of lactic microflora of kefir grains and kefir beverage by using culture-dependent and culture-independent methods

Abstract: In this study, we investigated the bacterial compositions of kefir grains and kefir beverages collected from different regions of Turkey by using culture‐independent and culture‐dependent methods. In the culture‐independent detection, 10 different species of bacteria were detected in total by using the polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE) analysis of the 16S rRNA gene V3 region. Among these species, Lactobacillus kefiranofaciens was the most dominant one in the kefir grains, while Lactococcus lactis was found to be significantly prevalent in the kefir beverages. In the culture‐dependent detection, the primary differentiation and grouping of the isolates from kefir beverages and kefir grains were performed using repetitive sequence‐based PCR (rep‐PCR) fingerprinting, and the results were validated by 16S rDNA full‐length sequencing. According to the results of culture‐dependent methods, the most frequently isolated species were L. lactis, Leuconostoc mesenteroides, and Lactobacillus kefiri, respectively. Only 3 species, which are L. lactis, Lactobacillus acidophilus, and Streptococcus thermophilus, were detected with both culture‐dependent and culture‐independent methods. This study showed that the combination of both methods is necessary for a detailed and reliable investigation of microbial communities in kefir grains and kefir beverages. Practical Application: Due to their artisan‐ and region‐dependent microflora, kefir products can be a source of interesting lactic acid bacteria, either new taxa or strains with specific functional properties, which might be used for the development of new starter cultures and innovative food products. Therefore, an increasing demand exists for new strains that show desirable effects on the product characteristics Artisan dairy products are a candidate source of such microorganisms. For this reason, in this study, the bacterial compositions of kefir grains and kefir beverages obtained from different regions of Turkey were studied using culture‐dependent and culture‐independent molecular methods.     

Comparative study of the phenolics,antioxidant and metagenomic composition of novel soy whey-based beverages produced using three different water kefir microbiota

Water kefir microbiota was used to develop novel soy whey-based beverages that have antioxidant activity. In the present study, comparative phenolics, antioxidant and metagenomic composition of the soy whey beverages fermented using three different water kefir microbiota, named WKFS-A, WKFS-B and WKFS-C were investigated. WKFS-B beverage had the highest concentrations of isoflavone aglycones (208.73 ± 2.78 mg L⁻¹) and phenolic acids (132.33 ± 3.41 mg L⁻¹) compared with WKFS-A (193.88 ± 1.15 mg L⁻¹) and (91.73 ± 2.34 mg L⁻¹) and WKFS-C (160.63 ± 1.76 mg L⁻¹) and (97.13 ± 2.63 mg L⁻¹), respectively. The WKFS-B also showed higher DPPH and ABTS radical scavenging activity and ferric reducing antioxidant power compared with WKFS-A and WKFS-C beverages. Microbial species diversity index analysis showed that a higher concentration of isoflavone aglycones, phenolic acids and increased antioxidant activity in the WKFS-B beverage correlates with the higher relative abundance of Lactobacillus genus. This study thus revealed that Lactobacillus dominated water kefir microbiota produces soy whey beverages with high phenolic acids, isoflavone aglycones and antioxidant activity.     

Kefir grains as a starter for whey fermentation at different temperatures: chemical and microbiological characterisation

We report here a comparative analysis of the growth, acidification capacity, and chemical and microbiologic composition between kefir grains after 20 subcultures in whey at 20, 30, and 37°C and the original kefir grains coming from milk along with a determination of the microbiological composition of the fermented whey as compared with that of traditional fermented milk. When fermentation was carried out repeatedly at 30 or 37°C, kefir grains changed their kefir-like appearance, exhibited reduced growth rates, had a lower diversity of yeasts and water content, and a higher protein-to-polysaccharide ratio compared with the original kefir grains. In contrast, at 20°C kefir grains could remain in whey for prolonged periods without altering their acidification capacity, growth rate, macroscopic appearance or chemical and microbiologic composition-with the only difference being a reduction in certain yeast populations after 20 subcultures in whey. At this incubation temperature, the presence of Lactobacillus kefiranofaciens, Lb. kefir, Lb. parakefir, Lactococcus lactis, Kluyveromyces marxianus, Saccharomyces unisporus, and Sac. cerevisiae was detected in kefir grains and in fermented whey by denaturing-gradient-gel electrophoresis (DGGE). In whey fermented at 20°C the number of lactic-acid bacteria (LAB) was significantly lower (P<0·05) and the number of yeast significantly higher (P<0·05) than in fermented milk. Since the DGGE profiles were similar for both products, at this temperature the microbiologic composition of fermented whey is similar to that of fermented milk. We therefore suggest a temperature of 20°C to preserve kefir grains as whey-fermentation starters.