Co-culturing Propionibacterium freudenreichii and Bifidobacterium animalis subsp. lactis improves short-chain fatty acids and vitamin B12 contents in soy whey.

The lack of vitamin B12 in unprocessed plant-based foods can lead to health problems in strict vegetarians and vegans. The main aim of this study was to investigate the potential synergy of co-culturing Bifidobacterium animalis subsp. lactis and Propionibacterium freudenreichii in improving production of vitamin B12 and short-chain fatty acids in soy whey. Different strategies including mono-, sequential and simultaneous cultures were adopted. Growth, short-chain fatty acids and vitamin B12 were assessed throughout the fermentation while free amino acids, volatiles, and isoflavones were determined on the final day. P. freudenreichii monoculture grew well in soy whey, whereas B. lactis monoculture entered the death phase by day 4. Principal component analysis demonstrates that metabolic changes in both sequential cultures did not show drastic differences to those of P. freudenreichii monoculture. However, simultaneous culturing significantly improved vitamin B12, acetic acid and propionic acid contents (1.3 times, 5 times, 2.5 times, compared to the next highest treatment [sequential cultures]) in fermented soy whey relative to other culturing modes. Hence, co-culturing of P. freudenreichii and B. lactis would provide an alternative method to improve vitamin B12, acetic acid and propionic acid contents in fermented foods.

Utilization of propionic acid bacteria in the biotransformation of soy (tofu) whey: Growth and metabolite changes.

Soy whey, a by-product from the tofu and soy protein isolate industry was evaluated as a substrate for a biofortified beverage using several propionic acid bacteria (PAB). PAB growth and changes in sugars, organic acids, amino acids and isoflavones were investigated. Vitamin B12 and short-chain fatty acid (SCFA) production were measured over time. Acidipropionibacterium acidipropionici (DSM 20272) showed the highest growth, compared to the other three PABs (Propionibacterium freudenreichii [DSM 20271 and DSM 4902], A. jensenii [DSM 20535]). Acidipropionibacterium (DSM 20272 and DSM 20535) showed the best propionic acid and acetic acid production, while P. freudenreichii produced the most succinic acid. Propionibacterium freudenreichii exhibited significant vitamin B12 production at 4.06 ± 0.28 µg/L for DSM 20271, followed by 2.58 ± 0.22 µg/L for DSM 4902. Notably, all PAB displayed strong ß-glycosidase activities evidenced by the conversion of isoflavone glycosides to isoflavone aglycones. The stark differences between Acidipropionibacterium spp. and Propionibacterium spp. indicate that the former PAB is specialized in SCFA production, while the latter PAB is better at vitamin B12 bioenrichment. This study demonstrated the possibility of employing PAB fermentation to improve SCFA and vitamin B12 content. This can open avenues for a beverage or functional ingredient development.

Evaluating the effect of lactic acid bacterial fermentation on salted soy whey for development of a potential novel soy sauce-like condiment.

There were two main objectives of this study: (1) to understand the effect of salt concentration on the growth of four lactic acid bacteria (LAB) in soy whey and determine the non-volatile and volatile profiles generated after fermentation; (2) to evaluate the potential of using salted soy whey to develop a sauce-like condiment through LAB fermentation. The four LAB included non-halophilic Lactiplantibacillus plantarum ML Prime, Limosilactobacillus fermentum PCC, Oenococcus oeni Enoferm Beta and halophilic Tetragenococcus halophilus DSM20337. At 2% salt, all LAB grew remarkably from day 0 to day 1, except for T. halophilus, while at 6% salt, the growth of L. plantarum, L. fermentum and O. oeni was suppressed. Conversely, the higher salt concentration enhanced the growth of T. halophilus in soy whey as the cell count only increased from 6.36 to 6.60 log CFU/mL at 2% salt but it elevated from 6.61 to 7.55 log CFU/mL at 6% salt. Similarly, the higher salt content negatively affected the sugar and amino acids metabolism and organic acids production by non-halophilic LAB. L. plantarum and O. oeni generated significantly (p < 0.05) more lactic acid (3.83 g/L and 4.17 g/L, respectively) than L. fermentum and T. halophilus (2.02 g/L and 0 g/L, respectively) at 2% salt. In contrast, a higher amount of acetic acid was generated by L. fermentum (0.72 g/L at 2% salt) and T. halophilus (0.51 g/L at 6% salt). LAB could remove the green and beany off-flavours in soy whey by metabolizing C6 and C7 aldehydes. However, to develop a novel soy sauce-like condiment, yeast fermentation and Maillard reaction may be required to generate more characteristic soy sauce-associated aroma compounds.

Exploring the feasibility of biotransforming salted soy whey into a soy sauce-like condiment using wine yeast Torulaspora delbrueckii and soy sauce yeasts Zygosaccharomyces rouxii and Candida versatilis as single starter cultures.

Salted soy whey, a liquid by-product from salted tofu processing, is a source of valuable nutrients. However, it is often under-utilized due to its high salt content. The objective of this study was to investigate the feasibility of using different species of yeast to transform salted soy whey into a soy sauce-like condiment. Three yeasts were used for salted soy whey biotransformation: Zygosaccharomyces rouxii NCYC 1682, Candida versatilis NCYC 1433, and Torulaspora delbrueckii Biodiva. This study focused on the growth of the yeasts in soy whey added with different levels of NaCl and the physicochemical changes of salted soy whey after fermentation. The soy sauce yeasts (Z. rouxii and C. versatilis) grew by approximately 2 log CFU/mL in soy whey with 2% and 10% salt while the cell count of wine yeast T. delbrueckii increased by around 1.5 log CFU/mL at 10% salt and 1.78 log CFU/mL at 2% salt after 14 days of fermentation. Candida versatilis grew better at 10% salt with less glucose consumption (consumed 68.49% at 10% versus 94.05% at 2% salt) than at 2% salt. It was also found that all three yeasts converted isoflavone glycosides (bound isoflavones) into aglycones (free isoflavones), the latter having better bioavailability. C. versatilis showed the greatest ability to transform isoflavone glycosides in salted soy whey into higher amounts of aglycones (conversion efficiency of 23.04% at 2% salt and 15.05% at 10% salt). Salted soy whey fermented with different yeasts had different volatile profiles. Soy sauce yeasts produced more isobutyl alcohol, isoamyl alcohol and volatile phenols while T. delbrueckii generated a substantial amount of ethanol and esters. This study revealed the growth and flavour modulating potential of yeasts in salted soy whey fermentation, which provides a possible avenue to develop a soy sauce-like condiment using salted soy whey as a substrate.

The impact of mixed amino acids supplementation on Torulaspora delbrueckii growth and volatile compound modulation in soy whey alcohol fermentation.

Soy (tofu) whey is a liquid side stream generated from tofu production and is often discarded as waste after it is generated. Direct disposal of soy whey can result in environmental issue in the long run. Soy whey has been previously successfully fermented using different types of wine yeasts, but the yeast available nitrogen (YAN) was found to be deficient. In this study, the soy whey YAN was estimated to be approximately 45.9 mg N/L. A mixture of four amino acids (valine, leucine, isoleucine and phenylalanine) was added into soy whey at a total concentration of +40, +80, +120 and +160 mg N/L and fermented with Torulaspora delbrueckii Biodiva for a period of 10 days. Increasing amino acid supplementation did not affect the yeast cell growth, but it sped up the sugar utilization proportionally. Increasing amino acid supplementation resulted in lower organic acid production and higher glycerol production. Amino acid supplementation also enhances the production rate of higher alcohols; increasing amount of higher alcohols and their respective esters were obtained with increasing amount of amino acid supplementation. However, higher levels of amino acid supplementation (particularly at +160 mg N/L sample) resulted in higher residual nitrogen contents which may lead to microbial instability. Supplementation of 120 mg N/L of amino acids was found to be the optimum concentration to enhance the metabolism of the yeast without leaving a high residual amino acid content. Therefore, with proper control of the amino acid addition dosage, the usage of mixed amino acid supplementation may be a strategy to regulate the fermentation kinetics and volatile compound modulation in soy whey alcohol fermentation.

Understanding the interaction of isoleucine paired with other amino acids in soy whey alcohol fermentation using Torulaspora delbrueckii.

Soy (tofu) whey is a liquid by-product generated from tofu (soybean curd) production and it is often discarded off as a waste liquid by the tofu manufacturers. Previous studies have demonstrated that soy whey can be biotransformed into a soy alcoholic beverage by using Saccharomyces and non-Saccharomyces yeasts even though soy whey is low in yeast assimilable nitrogen (YAN) content. In this study, the initial YAN of the soy whey was estimated to be 46.6 mg N/L and Torulaspora delbrueckii Biodiva was used to ferment soy whey supplemented with either isoleucine only or isoleucine paired with valine, leucine or phenylalanine (each amino acid supplemented at a dosage of 30 mg N/L). Amino acid supplementation was found to enhance sugar utilization by the yeast, which led to higher ethanol production (7.49% v/v in control versus 8.35-8.80% v/v in supplemented samples). Samples supplemented with isoleucine only experienced slower sugar utilization during the fermentation as compared to the paired amino acid samples, but the yeast was still able to utilize the sugar to low levels at the end of the fermentation. The presence of leucine supplementation counteracted the "inhibition" induced by the presence of isoleucine at the first day of the fermentation. Amino acid supplementation slowed down glutamic acid utilization and resulted in higher levels of residual glutamic acid and alanine. Amino acid supplementation increased the corresponding fusel alcohol production and the presence of other amino acids reduced the active amyl alcohol production. Therefore, interactions between amino acids can impact the metabolism of the yeast as well as the flavor modulation during soy whey fermentation.

Effect of single amino acid addition on growth kinetics and flavor modulation by Torulaspora delbrueckii in soy (tofu) whey alcoholic beverage fermentation.

Soy (tofu) whey is a by-product commonly disposed of by tofu manufacturers around the world. Due to its nutritious nature, direct disposal of soy whey into the sewage can result in a detrimental impact on the environment in the long-run. In this study, soy whey supplemented with four individual amino acids (valine, leucine, isoleucine and phenylalanine) equivalent to 120 mg N/L was fermented with a yeast Torulaspora delbrueckii Biodiva. The supplementation of an individual amino acid resulted in faster sugar utilization and lower levels of residual sugar than the unsupplemented whey but did not result in a significantly higher amount of ethanol (7-8% v/v) at the end of fermentation. Isoleucine supplementation resulted in a slightly slower initial yeast growth rate when compared to the control while the other three amino acids had identical yeast growth kinetics to the control. Isoleucine supplementation also resulted in slower sugar utilization during the first four days. Therefore, isoleucine is least preferred by the yeast. The supplementation of amino acids resulted in greater formation of higher alcohols and their corresponding alcohol-derived esters. Overall, the supplementation of a single amino acid enhanced sugar utilization and impacted flavor compounds of the resultant soy whey alcoholic beverage.

Survival of probiotic strain Lactobacillus paracasei L26 during co-fermentation with S. cerevisiae for the development of a novel beer beverage.

Amidst the rising popularity of craft beers, it would be opportune to develop a novel, unfiltered and unpasteurized sour beer with high probiotic live counts. However, as beer typically contains hop iso-α-acids that prevent the growth and survival of probiotic lactic acid bacteria, the use of suitable fermentation strategies is crucial. The growth, and survival of the probiotic bacterium, Lactobacillus paracasei L26, were assessed during a 10-day co-fermentation period with a brewer's yeast, Saccharomyces cerevisiae S-04, in unhopped wort. Isomerized hop extract was added prior to storage of the beers at 25 °C and 5 °C. During co-fermentation in unhopped wort, L. paracasei L26 maintained high viable cell counts above 8 Log CFU/mL, indicating species compatibility with the yeast. The majority of fermentable sugars were attenuated by S. cerevisiae S-04, with a concomitant production of alcohols and esters. Significant amounts of lactic acid were produced by L. paracasei L26 (P < 0.05). During storage with added isomerized hop extract, maximal probiotic viability enhancing effects were observed in the presence of live S. cerevisiae S-04, in combination with refrigeration. The results suggest that beers could be a vehicle for probiotic delivery under appropriate conditions. This was the first study demonstrating the feasibility of utilizing probiotic lactobacilli as starter cultures in beer brewing.

Evaluation of five commercial non-Saccharomyces yeasts in fermentation of soy (tofu) whey into an alcoholic beverage.

Soy (tofu) whey is a nutritious liquid substrate that is often discarded by tofu manufacturers. Recent research has shown that tofu whey can be converted into a soy alcoholic beverage using Saccharomyces yeasts. In this study, five commercially available non-Saccharomyces yeasts (Torulaspora delbrueckii; Lachancea thermotolerans; Metschnikowia pulcherrima; Pichia kluyveri and Williopsis saturnus) were evaluated in tofu whey fermentation and each of the yeasts showed different growth kinetics and fermentation performance. T. delbrueckii and L. thermotolerans consumed the supplemented sucrose and produced 6-7% (v/v) ethanol, while M. pulcherrima, P. kluyveri and W. saturnus only utilized the endogenous fructose and glucose, producing trace levels of ethanol. Besides, different yeasts showed different ß-glucosidase activities with 22-97% reduction in isoflavone glucosides; T. delbrueckii, L. thermotolerans and W. saturnus also decreased the level of GABA in tofu whey. Endogenous volatile compounds (mainly short-chain aldehydes) in tofu whey were reduced to trace levels, but different volatile compounds were produced by different yeasts at varying levels that can contribute to the different aroma profiles of the beverages. Therefore, selection of appropriate non-Saccharomyces yeasts can serve as a new strategy to valorize tofu whey and alter the aroma profile of the beverage.

The Possible Reduction Mechanism of Volatile Sulfur Compounds during Durian Wine Fermentation Verified in Modified Buffers.

Durian fruit is rich in volatile sulfur compounds (VSCs), especially thiols and disulfides, which contribute to its onion-like odor. After fermentation, these VSCs were reduced to trace or undetectable levels in durian wine. The possible reduction mechanism of these VSCs (especially diethyl disulfide and ethanethiol) was investigated in a modified buffer in the presence of sulfite at different pH. An interconversion between diethyl disulfide and ethanethiol was found to be dependent on the pH: the higher the pH, the higher production of ethanethiol. It is suggested that, during durian wine fermentation, disulfides endogenous to durian pulp might be firstly converted into their corresponding thiols in the presence of reductant sulfite formed by yeast. The produced thiols as well as the thiols endogenous to the durian pulp were then removed by the mannoproteins of yeast lees.

Biotransformation of soy whey into soy alcoholic beverage by four commercial strains of Saccharomyces cerevisiae.

Soy whey is a liquid waste stream generated from tofu and soy protein manufacturing, and is commonly disposed of into the drainage system in food industry. Instead of disposing of soy whey as a waste, it could be used to produce alcoholic beverages. This study investigated the feasibility of converting soy whey into soy alcoholic beverage using four commercial Saccharomyces cerevisiae strains as a zero-waste approach to tackle the soy whey disposal issue. The four Saccharomyces yeasts grew by approximately 2logCFU/mL and produced approximately 7-8% (v/v) of ethanol. Isoflavone glucosides were hydrolyzed and transformed into isoflavone aglycones, increasing the antioxidant capacity. New aroma-active volatiles, especially esters and higher alcohols, were produced and imparted fruity and floral notes to the soy alcoholic beverage. Therefore, alcoholic fermentation would serve as a solution toward zero-waste manufacturing by biotransforming soy whey into a world's first novel functional alcoholic beverage naturally enriched with free isoflavones.

The effects of co- and sequential inoculation of Torulaspora delbrueckii and Pichia kluyveri on chemical compositions of durian wine.

This is a first study on using two non-Saccharomyces yeasts, Torulaspora delbrueckii Biodiva and Pichia kluyveri FrootZen to produce durian wine via co-inoculation (Co-I) and sequential inoculation (Seq-I). T. delbrueckii inhibited the growth of P. kluyveri and P. kluyveri also partly retarded the growth of T. delbrueckii in Co-I and Seq-I treatments. Co-I and Seq-I produced similar levels of ethanol to T. delbrueckii Biodiva monoculture. In addition, Seq-I increased malic acid degradation and higher succinic acid production. Compared with T. delbrueckii Biodiva, Co-I produced similar amounts of ethyl esters, higher alcohols and moderately increased levels of ethyl acetate. Seq-I 2th (T. delbrueckii inoculated after 2 days fermentation with P. kluyveri) and Seq-I 5th produced excessive amounts of ethyl acetate (≥ 80 mg/L) but relatively lower levels of higher alcohols. This study suggested that Co-I could complete alcoholic fermentation with more complex aromas and might be novel way for wine making.

Chemical consequences of three commercial strains of Oenococcus oeni co-inoculated with Torulaspora delbrueckii in durian wine fermentation.

This work evaluated for the first time the chemical consequences of three commercial strains of Oenococcus oeni co-inoculated with Torulaspora delbrueckii in durian wine fermentation. Compared with the control (yeast only, 5.70% v/v ethanol produced), samples co-inoculated with T. delbrueckii and O. oeni PN4 improved ethanol production (6.06% v/v), which was significantly higher than samples co-inoculated with Viniflora (4.78% v/v) or Enoferm Beta (5.01% v/v). Wines co-fermented with the respective latter two oenococci contained excessive levels of ethyl acetate (>80mg/L) that were likely to affect negatively wine aroma. In addition, they led to significantly higher acetic and lactic acid production relative to PN4. O. oeni PN4 seemed to be the most suitable strain to co-inoculate with T. delbrueckii for simultaneous alcoholic and malolactic fermentation in durian wine by contributing moderately increased concentrations of higher alcohols, acetate esters and ethyl esters that would have positive sensory impacts.

Biotransformation of chemical constituents of durian wine with simultaneous alcoholic fermentation by Torulaspora delbrueckii and malolactic fermentation by Oenococcus oeni.

This work represents the first study on the biotransformation of chemical constituents of durian wine via simultaneous alcoholic fermentation (AF) and malolactic fermentation (MLF) with non-Saccharomyces yeast and lactic acid bacteria (LAB), namely, Torulaspora delbrueckii Biodiva and Oenococcus oeni PN4. The presence of PN4 improved the utilization of sugars but did not affect ethanol production. MLF resulted in the significant degradation of malic acid with corresponding increases in pH and lactic acid. The final concentrations of acetic acid (1.29 g/L) and succinic acid (3.70 g/L) in simultaneous AF and MLF were significantly higher than that in AF (1.05 and 1.31 g/L) only. Compared with AF, simultaneous AF and MLF significantly elevated the levels of aroma compounds with higher levels of higher alcohols (isoamyl alcohol, active amyl alcohol, isobutyl alcohol, and 2-phenylethyl alcohol), acetate esters (ethyl acetate, isoamyl acetate), and ethyl esters (ethyl octanoate, ethyl dodecanoate). All the endogenous volatile sulfur compounds decreased to trace or undetectable levels at the end of fermentation. MLF accentuated the reduction of acetaldehyde and sulfides. The initially absent dipropyl disulfide was formed, then catabolized, especially in simultaneous AF and MLF. This study suggested that the simultaneous AF and MLF of non-Saccharomyces and LAB could modify the volatile compositions and potentially modulate the organoleptic properties of durian wine.