Antagonism between Saccharomyces cerevisiae and Williopsis mrakii and significance for mixed yeast alcoholic beverage fermentations

This research was carried out to investigate the antagonism between Saccharomyces cerevisiae and Williopsis saturnus var. mrakii. When glucose level was ≥12% (w/v) (potential ethanol ≥7.2% v/v), W. mrakii that was co‐inoculated with S. cerevisiae died early, while W. mrakii alone could not survive in broth with ≥8% (v/v) ethanol. Moreover, when glucose was ≥15% (w/v) (potential ethanol ≥9.0% v/v), higher inoculums of S. cerevisiae induced faster cell decline of W. mrakii; when glucose was ≤2% (w/v), there was no early cell decline of W. mrakii at any inoculum ratio, but there was inoculum ratio‐dependent growth retardation. These results indicate that ethanol was at least one of the factors causing early cell decline of W. mrakii in co‐culture with S. cerevisiae. Further, S. cerevisiae was inhibited when it was inoculated sequentially into a W. mrakii monoculture; the former yeast was also suppressed when it was inoculated into the supernatants of a W. mrakii monoculture. This suggests that S. cerevisiae was sensitive to the mycosin (killer toxin) produced by W. mrakii. These findings have implications for controlling mixed yeast alcoholic beverage fermentations.     

水开菲尔粒发酵香蕉皮过程中成分变化研究

采用水开菲尔粒分别发酵蒸煮和未蒸煮香蕉皮,按红糖、香蕉皮、水、水开菲尔粒1:3∶10∶0.5的质量比于28 ℃发酵。 结果表明, 在发酵初期48 h内,发酵液的pH值及糖度均迅速下降,此后显著变缓。 240 h后测得未蒸煮和蒸煮（100 ℃蒸煮15 min）香蕉皮的利用 率分别为32.4%和43.8%;发酵液的pH值分别为3.53和3.51;滴定酸度分别为50.91 mmol/100 mL和54.89 mmol/100 mL;乙醇含量分 别为7.90 mg/mL和2.80 mg/mL;抗氧化能力（ORAC）值分别为5.03 mmol Trolox/L和3.58 mmol Trolox/L。 香蕉皮原料的挥发性成分主 要是丁酸酯类,发酵后这些成分全部消失,并产生大量新的挥发性成分,包括乙酸异戊酯、辛酸乙酯、芳樟醇等芳香成分。所得发酵液 色泽类似红茶水,风味类似果醋,具浓郁果香。 蒸煮香蕉皮适合用水开菲尔粒酿制果醋。     

Biochemical characteristics,virulence traits and antifungal resistance of two major yeast species isolated from kefir: Kluyveromyces marxianus and Saccharomyces unisporus

Kefir yeasts were investigated for their possible virulence using an in vitro system. Unweighted pair group method with arithmetic mean (UPGMA) cluster analysis of 46 biochemical traits showed that all kefir isolates formed clusters distinct from Candida albicans. No isolate showed N‐acetylglucosaminidase activity, which is involved in resistance against host immune systems. All Kluyveromyces marxianus isolates formed pseudohyphae and grew at 42 °C, whereas Saccharomyces unisporus isolates did not. None of the kefir yeasts had proteolytic or haemolytic activities. All K. marxianus isolates were susceptible to fluconazole, whereas S. unisporus isolates were classified as resistant.     

Quality characteristics of kefir as a carrier for probiotic Lactobacillus rhamnosus GG

Kefir was prepared using kefir grains of different origins (US, Australia and India) and starter culture. The sensory scores for kefir made using starter culture and US kefir grains were significantly higher than for the other samples (P < 0.05). The index of viscosity was significantly higher (P < 0.05) in kefir prepared using Indian kefir grains. Shear stress–shear rate plots indicated yield stress in all kefirs. All samples exhibited shear‐thinning behaviour. The average count of Lactobacillus rhamnosus GG in kefir after incubation was 7.09 log CFU/g. No significant difference (P > 0.05) was observed in quality characteristics in kefir after the addition of LGG, indicating that it could be a potential probiotic carrier.     

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.     

Effects of pulsed electric fields on the survival behaviour of Saccharomyces cerevisiae suspended in single solutions of low concentration

This paper investigates the survival behaviour of Saccharomyces cerevisiae cells suspended in NaCl solutions with various low concentrations after exposure to pulsed electric field (PEF) treatments. The input and heat‐generation pulse power during each PEF treatment experiment were measured directly. In addition, detailed information on sublethally PEF‐injured S. cerevisiae cells was obtained by the use of image analysis. Results showed that both the field intensity and the total treatment time had significantly affected the survival behaviour of S. cerevisiae cells, whereas the concentration of NaCl solution had less effect. When the S. cerevisiae cells were suspended in the solution with conductivity of 300 μS cm^?1, a high ratio of surviving cells was observed. Image analysis indicated a large number of sublethally injured cells (SICs) after the PEF treatment with low field intensity. Results indicate that the survival behaviour of cells suspended in solutions of low concentration was influenced by PEF intensity.     

The microbial diversity of water kefir

The microbial diversity of water kefir, made from a mixture of water, dried figs, a slice of lemon and sucrose was studied. The microbial consortia residing in the granules of three water kefirs of different origins were analyzed. A collection of 453 bacterial isolates was obtained on different selective/differential media. Bacterial isolates were grouped with randomly amplified polymorphic DNA (RAPD)-PCR analyses. One representative of each RAPD genotype was identified by comparative 16S rDNA gene sequencing. The predominant genus in water kefirs I and II was Lactobacillus, which accounted for 82.1% in water kefir I and 72.1% in water kefir II of the bacterial isolates. The most abundant species in water kefirs I and II were Lactobacillus hordei and Lb. nagelii followed by considerably lower numbers of Lb. casei. Other lactic acid bacteria (LAB) were identified as Leuconostoc mesenteroides and Lc. citreum in all three water kefirs. The most abundant species in water kefir III was Lc. mesenteroides (28%) and Lc. citreum (24.3%). A total of 57 LAB belonging to the species of Lb. casei, Lb. hordei, Lb. nagelii, Lb. hilgardii and Lc. mesenteroides were able to produce exopolysacchrides from sucrose. Non LABs were identified as Acetobacter fabarum and Ac. orientalis. The Acetobacter species were more prevalent in consortium III. Cluster analyses of RAPD-PCR patterns revealed an interspecies diversity among the Lactobacillus and Acetobacter strains. Aditionally, Saccharomyces cerevisiae, Lachancea fermentati, Hanseniaospora valbyensis and Zygotorulaspora florentina were isolated and identified by comparison of partial 26S rDNA sequences and FTIR spectroscopy.     

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.     

Acid adaptation for improvement of viability of Saccharomyces cerevisiae during freeze‐drying

This work was aimed to investigate its adaptation to moderate acid stress and the resulting improvement in its viability during freeze‐drying. When Saccharomyces cerevisiae UP3OY5 strain was adapted to acid condition (pH 3.5), the viability of acid‐adapted cells (79.9%) was significantly higher than that of control cells (40.5%) after freeze‐drying with trehalose as a carrier. Membrane fatty acid profile of acid‐adapted cells changed significantly in comparison with that of control cells. An increase in fatty acid saturation degree that led to 1.76‐fold increase in the ratio of saturated to unsaturated fatty acids was shown. Intracellular glycogen content was found to be higher than that of control cells. On the contrary, the trehalose content of acid‐adapted cells was found to be much smaller than that of control cells. The key role of acid adaptation in acquiring cross‐protection mechanism was suggested to permit yeast to better survive to freeze‐drying.     

Effects of sugar concentration on mango wine composition fermented by Saccharomyces cerevisiae MERIT.ferm

This study investigated the effect of initial sugar concentrations (^°Brix of 17, 23 and 30) on mango wine composition fermented by Saccharomyces cerevisiae MERIT.ferm. It was found that growth rate and maximum cell population were inversely correlated with initial sugar levels with the fastest growth rate and largest cell population in the low sugar fermentation. However, the cell population in the low and medium sugar fermentation declined significantly (from 8.7 and 8.2 log to 2 and 2.8 log, respectively) relative to the high sugar fermentation in which cell populations remained stable upon reaching the stationary phase (7.7 log). Glycerol production increased with increasing sugar content in low (13.4 g L^?1), medium (14.5 g L^?1) and high (15.9 g L^?1) sugar fermentation. In addition, high sugar fermentation had a negative impact on volatile production with significantly lower amounts of acetate esters (1.5 mg L^?1) but more acetic acid (0.54 g L^?1) compared to the low (5.0 mg L^?1 and 0.44 g L^?1, respectively) and medium (3.7 mg L^?1 and 0.49 g L^?1, respectively) sugar fermentations. Furthermore, volatiles especially terpene hydrocarbons (α‐caryophyllene was released) present in mango juice were significantly metabolised after fermentation, while numerous new volatile compounds (such as isobutyl alcohol, isoamyl alcohol, ethyl and acetate esters) were produced. Some terpene alcohols were released and converted into corresponding acetyl esters. This may indicate that the mango wines fermented with different levels of sugars would have different flavour aromas.     

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.     

藏灵菇牦牛酸乳发酵过程中乳酸菌和酵母菌对品质的作用

藏灵菇酸乳是由乳酸菌、酵母菌、醋杆菌和霉菌共同发酵形成的一种发酵乳制品,其品质特征与普通酸乳有显著差异。为研究藏灵菇发酵酸乳中乳酸菌和酵母菌分别发挥的作用,利用放线菌酮和青链霉素分别抑制发酵过程中的酵母菌和乳酸菌,测定并比较正常发酵酸乳、抑制乳酸菌发酵酸乳和抑制酵母菌发酵酸乳的风味、口感和质构等相关指标。实验结果显示抑制酵母菌发酵的酸乳中乙醇含量由1.85g/kg上升为2.77g/kg,VB2含量由168μg/100g下降为157μg/100g,与正常发酵组差异较大;抑制乳酸菌发酵的酸乳中酸度由18°T上升为28°T,氨基酸态氮由65mg/100g下降为53mg/100g,乳酸含量持续2mg/g,乙酸由1mg/g下降为0.7mg/g,VB1含量由20μg/100g下降为18μg/100g,与正常发酵组差异较大。乳酸菌在酸乳发酵过程中,对酸度、氨基酸态氮、质构、乳酸、乙酸与VB1的生成贡献较大;酵母菌对乙醇、VB2的生成贡献较大。     

The effects of ratio of cow's milk to soymilk,probiotic strain and fruit concentrate on qualitative aspects of probiotic flavoured fermented drinks

The effects of proportion of cow's milk to soymilk (100:0, 75:25, 50:50, 25:75, 0:100), probiotic bacteria (Lactobacillus acidophilus LA‐5 or Lactobacillus casei L‐01) and natural fruit concentrates (strawberry, apricot, peach and pear) on quality characteristics of soy‐based probiotic drink were investigated. The parameters were analysed at the end of fermentation and during 21 days of storage at 5 ^°C. The highest viability was observed when the equal proportion of cow's milk and soymilk and L. casei was used (50:50‐CY). During chilled storage, the flavouring apricot had the highest stimulatory effect on the survival of L. casei in 50:50 treatment. In general, the treatment 50:50‐CY was realised as the best one overall.     

The probiotic role of Lactobacillus plantarum in reducing risks associated with cardiovascular disease

A novel strain of Lactobacillus plantarum DMDL 9010 (CGMCC No. 5172) was isolated from naturally fermented mustard. The potential cholesterol reduction effects of this strain were investigated using an in vivo model. The results showed that L. plantarum DMDL 9010 at a dose of 10⁹ cells per day significantly reduced (P ≤ 0.05) serum total cholesterol (TC), low‐density lipoprotein cholesterol content (LDL‐C) levels and atherosclerosis index (AI) by 23.03%, 28.00% and 34.03%, respectively, while L. plantarum DMDL 9010 did not exhibit a significant effect on reducing serum triglycerides (TG) and increasing the serum high‐density lipoprotein cholesterol content (HDL‐C) in experimental rats (P > 0.05). The morphological and pathological changes in the liver illustrated that L. plantarum DMDL 9010 protected the rats against hepatocyte steatosis. Additionally, a high dose of L. plantarum DMDL 9010 was shown to exhibit a positive cholesterol‐lowering effect through decreasing the liver cholesterol (?33.20%) and triglyceride (?40.86%) levels, and increasing significantly (P ≤ 0.05) faecal cholesterol (+31.07%) and bile acid excretion (+70.18%). The results demonstrated that L. plantarum DMDL 9010 acted in a dose‐dependent way to decrease serum and total hepatic cholesterol and triglyceride and enhance faecal excretion of bile acids. In conclusion, these findings suggest that L. plantarum DMDL 9010 has potential to be explored as a probiotic for hypercholesterolaemic preventive and therapeutic.     

Restarting incomplete fermentations: the effect of high concentrations of acetic acid

A high concentration of acetic acid has been suggested to be a cause of incomplete fermentation. Acetic acid can reduce the growth and fermentation activity of Saccharomyces cerevisiae , and incomplete ferments can contain a high concentration of acetic acid, although the origin is often not clear. Here we examined the concentration of acetic acid which could hinder or prevent the restarting of an incomplete Cabernet Sauvignon fermentation by three acclimatized wine yeast cultures. By taking advantage of new technology which combines reverse osmosis and ion exchange, the concentration of acetic acid in an incomplete commercial ferment was adjusted to between 0.5 and 4.0 g/L. At a concentration up to 4 g/L, acetic acid did not prevent the restarting of the incomplete ferment. No lag period was observed in any of the fermentations. The amount of sugar fermented after 20–21 days was dependent on the strain of Sacch. cerevisiae and was correlated with the initial concentration of acetic acid at the time of restarting, between 0.5 and 2.0 g/L, with the concentration of residual sugar increasing with the concentration of acetic acid. In ferments which contained 4 g acetic acid/L, however, the rate of sugar consumption was sufficiently low after 20–21 days to suggest that complete fermentation would not occur, even with prolonged incubation. Effective treatment of the incomplete fermentation was therefore possible in the presence of up to 2 g acetic acid/L. Cell viability and fermentation kinetics were also strongly dependent on the strain of yeast.     

A new synbiotic dairy food containing lactobionic acid and Lactobacillus casei

As well as its beneficial health properties, the incorporation of the prebiotic lactobionic acid (LBA) in fermented dairy products can provide a technological advantage due to its gelling capacities. This study aimed to develop a new functional dairy product containing LBA synthesised within the process itself by the bacterium Pseudomonas taetrolens. A probiotic Lactobacillus casei strain was introduced through a sequential fermentation system. After incubation, in the case of the most effective experimental procedure, a synbiotic fermented milk with 30 g/L of prebiotic LBA was obtained, together with an active L. casei population of 10⁹ cfu/mL and <1% lactose content.