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.     

Fermented milk obtained with kefir grains as an ingredient in breadmaking

The objectives of this research were to study the effect of the addition of lyophilised kefir milk to premixes for household production of bread and evaluate the quality attributes of them. Four lyophilised samples were obtained from the followings: skim milk, acidified skim milk, fermented skim milk and neutralised fermented skim milk. Breads were prepared with commercial wheat flour, lyophilised milk samples and yeast through a straight dough process. Quality was assessed through loaf volume, crumb porosity and moisture, crumb texture and crust colour. Changes in texture and starch recrystallisation by X‐ray diffractometry were determined after 1 and 3 days of storage at room temperature. Breads with acidified milks showed the highest specific volumes and crumbs with the best texture properties. Crystallinity in bread with fermented milks was higher than for skim milk sample. This would indicate that there would be a certain effect of the type of milk processing on the promotion of starch retrogradation.     

Identification and characterization of a glucan-producing enzyme from Lactobacillus hilgardii TMW 1.828 involved in granule formation of water kefir

Water kefir is a home made fermented beverage based on a sucrose solution with fruit extracts. The inoculum of such fermentations consists of macroscopic granula containing lactic and acetic acid bacteria, and yeasts, which are embedded in an exopolysaccharide (EPS) matrix. In this work, a strain of Lactobacillus hilgardii producing large amounts of the granule-forming dextran could be isolated. The glycosyltransferans (Gtf) commonly called glucansucrase responsible for the production of this dextran was purified from L. hilgardii. Characteristic enzyme kinetic data were obtained. Optimum activity was observed between pH 4.3 and 4.6 and temperatures between 40 °C and 45 °C. A Michaelis–Menten kinetic could be fit to the experimental data and a K_M of 0.0385 M was calculated. The corresponding gtf gene was identified and characterized. It encodes a 1448 amino acid protein with higher homologies to Gtfs of Lactobacillus parabuchneri, Lactobacillus sakei and Lactobacillus fermentum followed by lower homologies to Lactobacillus reuteri Gtfs. By knockout experiments the role of this gene in granule dextran production was demonstrated.     

Vitamin profiles of kefirs made from milks of different species

Kefirs were produced under laboratory conditions using 10 different kefir grain cultures from different households, and reconstituted cow, ewe, goat and mare milk as substrates. The base milks and corresponding kefirs were examined for their content of water-soluble vitamins and orotic acid. On average, enrichment of vitamin concentration of>20% was observed with thiamine (only ewe milk kefirs), pyridoxine (kefirs from all milk species except cow milk), and folic acid (kefirs from all milk species except mare milk). Orotic acid content was reduced during fermentation throughout.     

一种新型开菲尔奶制作工艺

将草莓汁、菠萝汁、白砂糖加入鲜牛奶中经杀菌冷却后接种开菲尔粒制作的发酵剂,25℃恒温发酵,采用正交试验法对其它工艺参数进行了研究.结果表明,草莓汁和菠萝汁的添加体积分数均为3%,白砂糖质量分数为8%,接种发酵剂体积分数4%,发酵16h后获得了一种口感、组织状态、风味俱佳的新型开菲尔奶.     

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.     

Short communication: Physicochemical features and microbial community of milk kefir using a potential probiotic Saccharomyces cerevisiae KU200284

The aim of this study was to analyze the β-glucan contents, physicochemical features, and microbial communities in milk kefir prepared using Saccharomyces cerevisiae KU200284 isolated from cucumber jangajji, a fermented vegetable commonly eaten in Korean. Three types of milk kefir were manufactured, with (1) activated kefir grain, (2) activated kefir grain with commercial S. cerevisiae BOF, and (3) activated kefir grain with S. cerevisiae KU200284. β-Glucan contents of milk kefir using kefir grain and kefir grain with S. cerevisiae strains BOF and KU200284 were 8.29, 8.59, and 8.57%, respectively. The pH, titratable acidity, viscosity, Brix level, and alcohol contents of milk kefir using kefir grain with S. cerevisiae strains were acceptable compared with milk kefir using only kefir grain. In milk kefir produced using kefir grains and S. cerevisiae strains, 16S rRNA reads showed representative strains of Lactobacillus kefiranofaciens (>72% relative abundance) and Acetobacter fabarum (>16% relative abundance). In particular, milk kefir using kefir grain with S. cerevisiae KU200284 had the highest relative abundance of L. kefiranofaciens. In addition, the internal transcribed sequence (ITS) rRNA reads in tested milk kefir showed representative strains of Kluyveromyces marxianus (>52% relative abundance) and Saccharomyces cerevisiae (>16% relative abundance). In contrast, milk kefir using S. cerevisiae strains had higher relative abundance of S. cerevisiae (>37%). The β-glucan production, physicochemical properties, and microbial community profiling indicate that S. cerevisiae KU200284 could be used in functional dairy products as a starter culture.     

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.     

An economic evaluation of freeze-dried kefir starter culture production using whey

An economic study is presented in which industrial-scale production of freeze-dried kefir starter culture is discussed based on results on a laboratory scale. Industrial scale-up was based on a 3-step process using 3 bioreactors of 100, 3,000, and 30,000 L for 300 kg of freeze-dried culture/d of plant capacity. The major cost component of the total investment was the freeze-drying machinery, which consisted of 57% of the total investment. Production cost was reduced from €15.4/kg ($18.5/kg) to ∉2.9/kg ($3.5/kg) when the production capacity was increased from 30 to 900 kg/d. An economic analysis revealed a 3.5-fold increase in production cost compared with the corresponding production cost of the wet product, with an added value of up to ∉10.8 × 10⁹ ($13.0 × 10⁹) within the European Union.