Fermentation

Most modern fermented foods and beverages are produced in fit-for-purpose facilities which are designed to ensure not only a reliable product, but also one safe for consumption. Despite careful hygiene, microorganisms can colonise these facilities and establish resident populations that can potentially contribute to the fermentation process. Although some microorganisms may not negatively affect the final product, spoilage microorganisms can be detrimental for quality, generating substantial economic losses. Here, amplicon-based phylotyping was used to map microbial communities within an Australian winery, before, during and after the 2020 vintage. Resident bacterial and yeast populations were shown to change over time, with both relative abundance and location within the winery varying according to sampling date. The bacterial family Micrococcaceae, and the genera Sphingomonas and Brevundimonas were the most abundant bacterial taxonomies, while Naganishia, Pyrenochaeta and Didymella were the most abundant fungal genera. Mapping the spatial distributions of the microbial populations identified the main locations that harboured these resident microorganisms, that include known wine spoilage yeasts and bacteria. Wine spoilage microorganisms, including the genefugura Lactobacillus, Acetobacter, Gluconobacter and Brettanomyces showed very low relative abundance and were found only in a couple of locations within the winery. Microbial populations detected in this facility were also compared to the resident microbiota identified in other fermented food facilities, revealing that microbial population structures may reflect the nature of the product created in each facility. Full article

The common use of commercial yeasts usually leads to dull wine with similar aromas and tastes. Therefore, screening for novel indigenous yeasts to practice is a promising method. In this research, aroma discrepancies among six wine groups fermentated with indigenous yeasts were analyzed. Three Saccharomyces yeasts (FS36, HL12, YT28) and three matched non-Saccharomyces yeasts (FS31, HL9, YT2) were selected from typical Chinese vineyards. The basic oenological parameters, aroma compounds, and sensory evaluation were analyzed. The results showed that each indigenous Saccharomyces yeast had excellent fermentation capacity, and mixed-strain fermentation groups produced more glycerol, contributing to sweeter and rounder taste. The results from GC-MS, principal components analysis (PCA), and sensory evaluation highlighted that the HL mixed group kept the most content of Marselan varietal flavors such as calamenene and β-damascone hereby ameliorated the whole aroma quality. Our study also implied that the indigenous yeast from the same region as the grape variety seems more conducive to preserve the natural variety characteristics of grapes. Full article

Adequate water-removal techniques are requisite to remain superior biofuel quality. The effects of vibrating types and operating time on the water-removal efficiency of molecular sieves were experimentally studied. Molecular sieves of 3 Å pore size own excellent hydrophilic characteristics and hardly absorb molecules other than water. Molecular sieves of 3 Å accompanied by two different vibrating types, rotary shaking and electromagnetic stirring, were used to remove initial water from the reactant mixture of feedstock oil in order to prevent excessive growth or breeding of microorganisms in the biofuel product. The physical structure of about 66% molecular sieves was significantly damaged due to shattered collision between the magnetic bar and molecular sieves during electromagnetic stirring for 1 h. The molecular sieves vibrated by the rotary shaker appeared to have relatively higher water-removal efficiency than those by the electromagnetic stirrer and by keeping the reactant mixture motionless by 6 and 5 wt.%, respectively. The structure of the molecular sieves vibrated by an electromagnetic stirrer and thereafter being dehydrated appeared much more irregular and damaged, and the weight loss accounted for as high as 19 wt.%. In contrast, the structure of the molecular sieves vibrated by a rotary shaker almost remained original ball-shaped, and the weight loss was much less after regenerative treatment for those molecular sieves. As a consequence, the water-removal process using molecular sieves vibrated by the rotary shaker is considered a competitive method during the biofuel production reaction to achieve a superior quality of biofuels. Full article

Strategies and actions for mitigating the emissions of greenhouse gas (GHG) and air pollutants in the transportation sector are becoming more important and urgent due to concerns related to public health and climate change. As a result, the Taiwanese government has promulgated a number of regulatory measures and promotion plans (or programs) on bioethanol use, novel fermentation research projects and domestic production since the mid-2000s. The main aim of this paper was to present a trend analysis of the motor gasoline supply/consumption and bioethanol supply, and the regulatory system relevant to bioethanol production and gasohol use since 2007 based on the official database and the statistics. The motor gasoline supply has shown a decreasing trend in the last five years (2016–2020), especially in 2020, corresponding to the impact of the COVID-19 outbreak in 2020. Although the government provided a subsidy of NT$ 1.0–2.0 dollars per liter for refueling E3 gasohol based on the price of 95-unleaded gasoline, the bioethanol supply has shown decreasing demand since 2012. In addition, the plans for domestic bioethanol production from lignocellulosic residues or energy crops were ceased in 2011 due to non-profitability. To examine the obstacles to bioethanol promotion in Taiwan, the bottlenecks to bioethanol production and gasohol use were addressed from the perspectives of the producer (domestic enterprise), the seller (gas station) and the consumer (end user). Full article

Selenium nanoparticles (SeNPs) are gaining importance in the food and medical fields due to their antibacterial properties. The microbial inhibition of these kinds of particles has been tested in a wide range of Gram (+) and Gram (−) pathogenic bacteria. When SeNPs are synthesized by biological methods, they are called biogenic SeNPs, which have a negative charge caused by their interaction between surface and capping layer (bioorganic material), producing their high stability. This review is focused on SeNPs synthesis by bacteria and summarizes the main factors that influence their main characteristics: shape, size and surface charge, considering the bacteria growth conditions for their synthesis. The different mechanisms of antimicrobial activity are revised, and this review describes several biosynthesis hypotheses that have been proposed due to the fact that the biological mechanism of SeNP synthesis is not fully known. Full article

4-Mercapto-4-methyl-2-pentanone (4MMP), a high-impact aroma compound with the box tree and black currant flavors was first identified in wines and could be released by microbial cysteine-S-conjugate β-lyases from its precursors. In this study, various yeasts and bacteria encoding β-lyases were selected to examine their β-lyase activities. A thiol precursor of 4MMP, cysteine-conjugate of 4MMP (cys-4MMP), was synthesized with a purity of >95% in a relatively environmentally friendly approach, and its chemical structure was confirmed by nuclear magnetic resonance spectroscopy. The β-lyase activities of the crude cell extract from the bacteria and yeast strains for different substrates were examined using a colorimetric method. Shewanella putrefaciens cell extract exhibited the highest β-lyase activity for all tested substrates. Additionally, the optimum pH and temperature for their β-lyase activities were determined. To monitor the conversion efficiency of precursor cys-4MMP to 4MMP, liquid chromatography-mass spectrometry was used. Our data indicate that selected bacteria and yeasts could convert cys-4MMP into 4MMP, and S. putrefaciens exhibited the best conversion yield. This study demonstrated the potential use of microbial cell extracts to produce sulfur-containing aroma compounds such as 4MMP. Full article

Manuka honey is known for its strong antibacterial effect against pathogens but can promote probiotic growth in certain conditions. In a two-factor ANOVA study, AMF^TM Manuka honey (Active Manuka Factor: 05⁺, 10⁺, 15⁺ and 20⁺) was utilised as a substrate for probiotic Limosilactobacillus reuteri DPC16 in an anaerobic batch fermenter for 36 h. The biomass growth in MRS broth was noticeably higher with AMF Manuka honey than invert syrup and control samples without any additional sweetener source. The pH value was significantly lowered below 4.0 only in the AMF samples with the formation of lactic acid as the major metabolite. Other beneficial short-chain fatty acids (SCFA), such as acetic, succinic, and propionic acids, produced during the fermentation, along with the honey saccharides, were quantified by two-dimensional (2-D) nuclear magnetic resonance (NMR) spectroscopy. A significantly (p < 0.05) high biomass in AMF 20⁺ sample after 36 h, can partly be attributed to the high total sugar and oligosaccharide content in the honey. Importantly, however, no statistically significant difference was observed in the recorded major fermentation outcomes for the different AMF levels. The results, nevertheless, indicate the potential prebiotic efficacy of Manuka honey as a fermentation substrate for the lactobacilli probiotic strain. Full article

Many contaminant yeast strains that survive inside fuel ethanol industrial vats show detrimental cell surface phenotypes. These harmful effects may include filamentation, invasive growth, flocculation, biofilm formation, and excessive foam production. Previous studies have linked some of these phenotypes to the expression of FLO genes, and the presence of gene length polymorphisms causing the expansion of FLO gene size appears to result in stronger flocculation and biofilm formation phenotypes. We performed here a molecular analysis of FLO1 and FLO11 gene polymorphisms present in contaminant strains of Saccharomyces cerevisiae from Brazilian fuel ethanol distilleries showing vigorous foaming phenotypes during fermentation. The size variability of these genes was correlated with cellular hydrophobicity, flocculation, and highly foaming phenotypes in these yeast strains. Our results also showed that deleting the primary activator of FLO genes (the FLO8 gene) from the genome of a contaminant and highly foaming industrial strain avoids complex foam formation, flocculation, invasive growth, and biofilm production by the engineered (flo8∆::Ble^R/flo8Δ::kanMX) yeast strain. Thus, the characterization of highly foaming yeasts and the influence of FLO8 in this phenotype open new perspectives for yeast strain engineering and optimization in the sugarcane fuel-ethanol industry. Full article

Inoculation of selected non-Saccharomyces yeasts with Saccharomyces cerevisiae as means to produce Merlot wines with reduced ethanol contents was investigated. Fermentations of grape musts (25.4° Brix, pH 3.50, and 4.23 g/L titratable acidity) were conducted in stainless steel tanks inoculated with Metschnikowia pulcherrima strains P01A016 or NS-MP or Meyerozyma guilliermondii P40D002 with S. cerevisiae Syrah added after three days. After fermentation, wines with Mt. pulcherrima contained 13.8% (P01A016) or 13.9% (NS-MP) v/v ethanol, respectively, amounts which were lower than in wines with S. cerevisiae alone (14.9% v/v). Delayed inoculation of must with S. cerevisiae (day 3) or musts with My. guilliermondii contained elevated concentrations of ethyl acetate (145 and 148 mg/L, respectively), concentrations significantly higher than those with S. cerevisiae inoculated on day 0 or with either strain of Mt. pulcherrima. Descriptive sensory analysis revealed a significant effect due to panelist but not due to Mt. pulcherrima or My. guilliermondii. This research indicates the potential for commercial application of these yeasts towards the production of reduced alcohol wines but without imparting negative sensory attributes. Full article

The microalga Chlorella and strains of Bifidobacterium have been used in human or animal food supplements for decades because of their positive health effects. The presented study assessed different properties of C. vulgaris and its combination with bifidobacteria with the aim to develop new functional foods. The growth of four bifidobacteria strains in milk and whey supplemented with 1.0% (w/v) C. vulgaris and the immunomodulatory effects of aqueous Chlorella solutions (0.5%, 1.0%, and 3.0%) on human peripheral mononuclear cells were evaluated. Furthermore, synergistic effects on lipid metabolism of rats fed a high-fat diet with Chlorella and B. animalis subsp. lactis BB-12^® were analysed. Chlorella had a positive growth-promoting effect on the tested bifidobacteria (p < 0.05), and significantly increased the secretion of inflammatory cytokines (tumor necrosis factor-α, interleukin-10, and interleukin-6), depending on the concentration of Chlorella (p < 0.05). After 8 weeks, significant synergistic effects of Chlorella and bifidobacteria on triglyceride levels in rat heart, liver, and serum were observed (p < 0.05). These results demonstrate that various combinations of Chlorella and bifidobacteria have significant potential for the development of new fermented products, dependent on the algal species, probiotic strain, application form, and concentrations for acceptable sensory quality for consumers. Full article

The selection of yeast strains adapted to fermentation stresses in their winegrowing area is a key factor to produce quality wines. Twelve non-Saccharomyces native strains from Denomination of Origin (D.O.) “Vinos de Madrid” (Spain), a warm climate winegrowing region, were tested under osmotic pressure, ethanol, and acidic pH stresses. In addition, mixed combinations between non-Saccharomyces and a native Saccharomyces cerevisiae strain were practised. Phenotypic microarray technology has been employed to study the metabolic output of yeasts under the different stress situations. The yeast strains, Lachancea fermentati, Lachancea thermotolerans, and Schizosaccharomyces pombe showed the best adaptation to three stress conditions examined. The use of mixed cultures improved the tolerance to osmotic pressure by Torulaspora delbrueckii, S. pombe, and Zygosaccharomyces bailii strains and to high ethanol content by Candida stellata, S. pombe, and Z. bailii strains regarding the control. In general, the good adaptation of the native non-Saccharomyces strains to fermentative stress conditions makes them great candidates for wine elaboration in warm climate areas. Full article

In this study, lactic acid bacteria (LAB) strains derived from human and fermented food sources were examined to identify their properties related to obesity, as well as establish their safety and stability as probiotics. LAB (Lacticaseibacillus rhamnosus MG4502, Lactobacillus gasseri MG4524, Limosilactobacillus reuteri MG5149, and Weissella cibaria MG5285) exhibited antioxidant activity through DPPH (>26.1%) and ABTS (>40.1%) radical scavenging assays and α-glucosidase inhibitory activities (>60.3%), respectively. The LAB strains promoted anti-adipogenesis by reducing lipid accumulation in 3T3-L1 cells by Oil Red O staining (>70.3%). In addition, we found that these LAB strains were resistant to simulated gastric and intestinal fluids (pH 3, 4, 7, and 8) and showed potential for health promotion, based on hemolysis, cell adhesion, antibiotic susceptibility, and enzyme production. Thus, LAB may be used as probiotic ingredients with beneficial effects. Full article

In connection with the active growth of poultry processing, the issue of rational use of poultry by-products is urgent. The paper proposes hydrolysis of the gizzards of broiler chickens and hens of the parent stock with bifidobacteria liquid concentrate (BLC) and Propionix liquid concentrated starter culture (Propionix LCSC). The effect of enzymatic treatment on changes in the structural components of the gizzards was studied using scanning electron microscopy (SEM) and determination of the dispersed composition. As a result of the research, the multiple regression equations and the response surfaces were obtained, which describe the optimal parameters of the gizzard´s hydrolysis process. The temperature and the time of hydrolysis are factors that have a significant effect on the degree of hydrolysis. The results of the structural and microscopic analysis confirm the high hydrolysability of the by-products by changing its structure and increasing the number of smaller protein particles. It was found experimentally that the gizzards of hens are more susceptible to the action of probiotic microorganisms’ enzymes compared to the gizzards of broiler chickens. Full article

Bored coffee beans (BCBs) are the residues left from the pest Hypothenemus hampei that attacks coffee crops, resulting in enormous economic losses. The bioconversion of monosaccharides from BCBs into hyaluronic acid (HA) is appealing both for using the residues and given the high commercial value of HA. This study dealt with the production of HA using Streptococcus zooepidemicus by employing either acid (AcH) or enzymatic (EnH) hydrolyzates from BCBs. The highest release of monosaccharides (evaluated using surface response methodology) was obtained with EnH (36.4 g/L); however, S. zooepidemicus produced more HA (1.5 g/L) using AcH compared to EnH. Hydrolyzates from acetone-extracted BCBs yielded 2.7 g/L of HA, which is similar to the amount obtained using a synthetic medium (2.8 g/L). This report demonstrates the potential of hydrolyzates from bored coffee beans to produce HA by S. zooepidemicus. Full article

The objective of this study was to determine the application of citric waste fermented yeast waste (CWYW) obtained from an agro-industrial by-product as a protein source to replace soybean meal (SBM) in a concentrate diet. We also determined the effect of various roughage to concentrate ratios (R:C) on the gas production kinetics, ruminal characteristics, and in vitro digestibility using an in vitro gas production technique. The experiment design was a 3 × 5 factorial design arranged in a completely randomized design (CRD), with three replicates. There were three R:C ratios (60:40, 50:50, and 40:60) and five replacing SBM with CWYW (SBM:CWYW) ratios (100:0, 75:25, 50:50, 25:75, and 0:100). The CWYW product’s crude protein (CP) content was 535 g/kg dry matter (DM). There was no interaction effect between R:C ratios and SBM:CWYW ratios for all parameters observed (p > 0.05). The SBM:CWYW ratio did not affect the kinetics and the cumulative amount of gas. However, the gas potential extent and cumulative production of gas were increased with the R:C ratio of 40:60, and the values were about 74.9 and 75.0 mL/0.5 g, respectively (p < 0.01). The replacement of SBM by CWYW at up to 75% did not alter in vitro dry matter digestibility (IVDMD), but 100% CWYW replacement significantly reduced (p < 0.05) IVDMD at 24 h of incubation and the mean value. In addition, IVDMD at 12 h and 24 h of incubation and the mean value were significantly increased with the R:C ratio of 40:60 (p < 0.01). The SBM:CWYW ratio did not change the ruminal pH and population of protozoa (p > 0.05). The ruminal pH was reduced at the R:C ratio of 40:60 (p < 0.01), whereas the protozoal population at 4 h was increased (p < 0.05). The SBM:CWYW ratio did not impact the in vitro volatile fatty acid (VFA) profile (p > 0.05). However, the total VFA, and propionate (C3) concentration were significantly increased (p < 0.01) by the R:C ratio of 40:60. In conclusion, the replacement of SBM by 75% CWYW did not show any negative impact on parameters observed, and the R:C ratio of 40:60 enhanced the gas kinetics, digestibility, VFA, and C3 concentration. Full article

The incidence and intensity of bushfires is increasing due to climate change, resulting in a greater risk of smoke taint development in wine. In this study, smoke-tainted and non-smoke-tainted wines were subjected to treatments using activated carbon with/without the addition of a cleaving enzyme treatment to hydrolyze glycoconjugates. Chemical measurements and volatile aroma compounds were assessed for each treatment, with the two smoke taint amelioration treatments exhibiting lower mean values for volatile aroma compounds exhibiting positive ‘fruit’ aromas. Furthermore, a low-cost electronic nose (e-nose) was used to assess the wines. A machine learning model based on artificial neural networks (ANN) was developed using the e-nose outputs from the unsmoked control wine, unsmoked wine with activated carbon treatment, unsmoked wine with a cleaving enzyme plus activated carbon treatment, and smoke-tainted control wine samples as inputs to classify the wines according to the smoke taint amelioration treatment. The model displayed a high overall accuracy of 98% in classifying the e-nose readings, illustrating it may be a rapid, cost-effective tool for winemakers to assess the effectiveness of smoke taint amelioration treatment by activated carbon with/without the use of a cleaving enzyme. Furthermore, the use of a cleaving enzyme coupled with activated carbon was found to be effective in ameliorating smoke taint in wine and may help delay the resurgence of smoke aromas in wine following the aging and hydrolysis of glycoconjugates. Full article

2,3-Butanediol (2,3-BD) is a reagent with remarkable commercial use as a platform chemical in numerous industries. The present study aims to determine the capabilities of non-pathogenic and cellulolytic Bacillus licheniformis 24 as a 2,3-BD producer. By applying the Plackett–Burman design and response surface methodology through central composite design (CCD), a complex optimization of medium and process parameters was conducted. Thus, among ten studied factors of medium content, four components were evaluated with a significant positive effect on 2,3-BD formation. Their optimal values for 2,3-BD production (yeast extract, 13.38 g/L; tryptone, 6.41 g/L; K₂HPO₄, 4.2 g/L; MgSO₄, 0.32 g/L), as well as the optimal temperature (37.8 °C), pH (6.23) and aeration rate (3.68 vvm) were predicted by CCD experiments and validated in a series of batch processes. In optimized batch fermentation of 200 g/L of glucose 91.23 g/L of 2,3-BD was obtained, with the overall productivity of 1.94 g/L/h and yield of 0.488 g/g. To reveal the maximum 2,3-BD tolerance of B. licheniformis 24, fed-batch fermentation was carried out. The obtained 138.8 g/L of 2,3-BD with a yield of 0.479 g/g and productivity of 1.16 g/L/h ranks the strain among the best 2,3-BD producers. Full article

Early detection of beer faults is an important assessment in the brewing process to secure a high-quality product and consumer acceptability. This study proposed an integrated AI system for smart detection of beer faults based on the comparison of near-infrared spectroscopy (NIR) and a newly developed electronic nose (e-nose) using machine learning modelling. For these purposes, a commercial larger beer was used as a base prototype, which was spiked with 18 common beer faults plus the control aroma. The 19 aroma profiles were used as targets for classification ma-chine learning (ML) modelling. Six different ML models were developed; Model 1 (M1) and M2 were developed using the NIR absorbance values (100 inputs from 1596–2396 nm) and e-nose (nine sensor readings) as inputs, respectively, to classify the samples into control, low and high concentration of faults. Model 3 (M3) and M4 were based on NIR and M5 and M6 based on the e-nose readings as inputs with 19 aroma profiles as targets for all models. A customized code tested 17 artificial neural network (ANN) algorithms automatically testing performance and neu-ron trimming. Results showed that the Bayesian regularization algorithm was the most adequate for classification rendering precisions of M1 = 95.6%, M2 = 95.3%, M3 = 98.9%, M4 = 98.3%, M5 = 96.8%, and M6 = 96.2% without statistical signs of under- or overfitting. The proposed system can be added to robotic pourers and the brewing process at low cost, which can benefit craft and larger brewing companies. Full article

During our search for ethanol-producing basidiomycete fungi for a wide range of substrates, we isolated Phlebia acerina, which is a white rot basidiomycete fungus. It favorably converted starch into ethanol with approximately 70% yield. Although the yield decreased as the starch concentration increased, growth and fermentation were observed even at 200 g/L of starch. P. acerina produced ethanol from glucose, galactose, mannose, xylose, cellobiose, and maltose with 93%, 91%, 86%, 72%, 92%, and 68% yields, respectively. Additionally, P. acerina, which secreted xylanase and xylosidase, was capable of assimilating xylan and directly converting it to ethanol with a yield of 63%. Furthermore, P. acerina produced ethanol directly from acorns, which are plant fruits containing starch and tannins, with a yield of 70%. Tannin delayed mycelia growth, thus prolonging ethanol production; however, this did not particularly affect the yield. These results were similar to those of fermentation in a medium with the same amounts of starch and tannin as the target crop acorn, thus suggesting that P. acerina could successfully produce environmentally friendly ethanol from starch-containing lignocellulosic biomass, unlike previously reported ethanol-producing basidiomycete fungi. Full article