Simultaneous inoculation of non-Saccharomyces yeast and lactic acid bacteria for aromatic kiwifruit wine production.

To further explore strain potential and develop an aromatic kiwifruit wine fermentation technique, the feasibility of simultaneous inoculation by non-Saccharomyces yeast and lactic acid bacteria was investigated. Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, and Limosilactobacillus fermentum, which have robust ß-glucosidase activity as well as good acid and ethanol tolerance, were inoculated for simultaneous fermentation with Zygosaccharomyces rouxii and Meyerozyma guilliermondii, respectively. Subsequently, the chemical compositions and sensory characteristics of the wines were comprehensively evaluated. The results showed that the majority of the simultaneous protocols effectively improved the quality of kiwifruit wines, increasing the content of polyphenols and volatile compounds, thereby enhancing sensory acceptability compared to the fermentation protocols inoculated with non-Saccharomyces yeast individually. Particularly, the collaboration between Lacp. plantarum and Z. rouxii significantly increased the diversity and content of esters, alcohols, and ketones, intensifying floral and seeded fruit odors, and achieving the highest overall acceptability. This study highlights the potential significance of simultaneous inoculation in kiwifruit wine production.

Use of Hanseniaspora guilliermondii and Hanseniaspora opuntiae to enhance the aromatic profile of beer in mixed-culture fermentation with Saccharomyces cerevisiae.

Beer production is predominantly carried out by Saccharomyces species, such as S. cerevisiae and S. pastorianus. However, the introduction of non-Saccharomyces yeasts in the brewing process is now seen as a promising strategy to improve and differentiate the organoleptic profile of beer. In this study, 17 non-Saccharomyces strains of 12 distinct species were isolated and submitted to a preliminary sensory evaluation to determine their potential for beer bioflavouring. Hanseniaspora guilliermondii IST315 and H. opuntiae IST408 aroma profiles presented the highest acceptability and were described as having 'fruity' and 'toffee' notes, respectively. Their presence in mixed-culture fermentations with S. cerevisiae US-05 did not influence attenuation and ethanol concentration of beer but had a significant impact in its volatile composition. Notably, while both strains reduced the total amount of ethyl esters, H. guilliermondii IST315 greatly increased the concentration of acetate esters, especially when sequentially inoculated, leading to an 8.2-fold increase in phenylethyl acetate ('rose', 'honey' aroma) in the final beverage. These findings highlight the importance of non-Saccharomyces yeasts in shaping the aroma profile of beer and suggest a role for Hanseniaspora spp. in improving it.

Study on the Sequential Combination of Bioethanol and Biogas Production from Corn Straw.

The objective of this study was to obtain two types of fuels, i.e., bioethanol and biogas, in a sequential combination of biochemical processes from lignocellulosic biomass (corn straw). Waste from the agricultural sector containing lignocellulose structures was used to obtain bioethanol, while the post-fermentation (cellulose stillage) residue obtained from ethanol fermentation was a raw material for the production of high-power biogas in the methane fermentation process. The studies on obtaining ethanol from lignocellulosic substrate were based on the simultaneous saccharification and fermentation (SSF) method, which is a simultaneous hydrolysis of enzymatic cellulose and fermentation of the obtained sugars. Saccharomyces cerevisiae (D-2) in the form of yeast cream was used for bioethanol production. The yeast strain D-2 originated from the collection of the Institute of Agricultural and Food Biotechnology. Volatile compounds identified in the distillates were measured using gas chromatography with flame ionization detector (GC-FID). CH4 and CO2 contained in the biogas were analyzed using a gas chromatograph in isothermal conditions, equipped with thermal conductivity detector (katharometer) with incandescent fiber. Our results show that simultaneous saccharification and fermentation enables production of bioethanol from agricultural residues with management of cellulose stillage in the methane fermentation process.

Simultaneous Saccharification and Fermentation of Sugar Beet Pulp with Mixed Bacterial Cultures for Lactic Acid and Propylene Glycol Production.

Research into fermentative production of lactic acid from agricultural by-products has recently concentrated on the direct conversion of biomass, whereby pure sugars are replaced with inexpensive feedstock in the process of lactic acid production. In our studies, for the first time, the source of carbon used is sugar beet pulp, generated as a by-product of industrial sugar production. In this paper, we focus on the simultaneous saccharification of lignocellulosic biomass and fermentation of lactic acid, using mixed cultures with complementary assimilation profiles. Lactic acid is one of the primary platform chemicals, and can be used to synthesize a wide variety of useful products, including green propylene glycol. A series of controlled batch fermentations was conducted under various conditions, including pretreatment with enzymatic hydrolysis. Inoculation was performed in two sequential stages, to avoid carbon catabolite repression. Biologically-synthesized lactic acid was catalytically reduced to propylene glycol over 5% Ru/C. The highest lactic acid yield was obtained with mixed cultures. The yield of propylene glycol from the biological lactic acid was similar to that obtained with a water solution of pure lactic acid. Our results show that simultaneous saccharification and fermentation enables generation of lactic acid, suitable for further chemical transformations, from agricultural residues.

Growth competition between lactic acid bacteria and Listeria monocytogenes during simultaneous fermentation and drying of meat sausages - A mathematical modeling.

Listeria monocytogenes is a significant foodborne health hazard in many products and may survive and grow when making fermented meat sausages. The objective of this study was to investigate the competition between lactic acid bacteria (LAB) and L. monocytogenes during simultaneous fermentation and drying (SFD) of meat sausages. Sausages made from irradiated ground beef (90% lean), salt, sugar, and sodium nitrite were inoculated with a 4-stain cocktail of LAB (2 Lactobacillus plantarum and 2 Lb. brevis strains) and a 5-strain cocktail of L. monocytogenes, individually or in combination, and incubated (30 °C, relative humidity 76%) for 5 days to undergo SFD. The changes in the populations of LAB and L. monocytogenes were monitored to determine the growth kinetics and examine the competitive growth between the two. L. monocytogenes grew in the sausage samples unhindered without LAB but was suppressed by LAB during SFD. The interaction between LAB and L. monocytogenes could be described by a modified Lotka-Volterra equation. The decreases of pH and aw in sausages could be related to the SFD time using segmented linear models. The competition model could accurately predict the growth of LAB and L. monocytogenes during SFD and may be used to improve the safety of semi-dry and dry fermented meat sausages.

Effects of Simultaneous Co-Fermentation of Five Indigenous Non-Saccharomyces Strains with S. cerevisiae on Vidal Icewine Aroma Quality.

In this study, Vidal grape must was fermented using commercial Saccharomyces cerevisiae F33 in pure culture as a control and in mixed culture with five indigenous non-Saccharomyces yeast strains (Hanseniaspora uvarum QTX22, Saccharomycopsis crataegensis YC30, Pichia kluyveri HSP14, Metschnikowia pulcherrima YC12, and Rhodosporidiobolus lusitaniae QTX15) through simultaneous fermentation in a 1:1 ratio. Simultaneous fermentation inhibited the growth of S. cerevisiae F33 and delayed the time to reach the maximum biomass. Compared with pure fermentation, the contents of polyphenols, acetic esters, ethyl esters, other esters, and terpenes were increased by R. lusitaniae QTX15, S. crataegensis YC30, and P. kluyveri HSP14 through simultaneous fermentation. S. crataegensis YC30 produced the highest total aroma activity and the most abundant aroma substances of all the wine samples. The odor activity values of 1 C13-norisoprenoid, 3 terpenes, 6 acetic esters, and 10 ethyl esters improved significantly, and three lactones (δ-decalactone, γ-nonalactone, and γ-decalactone) related to coconut and creamy flavor were only found in this wine. Moreover, this sample showed obvious "floral" and "fruity" note odor due to having the highest amount of ethyl ester aromatic substances and cinnamene, linalool, citronellol, ß-damascenone, isoamyl ethanoate, benzylcarbinyl acetate, isobutyl acetate, etc. We suggest that simultaneous fermentation of S. crataegensis YC30 with S. cerevisiae might represent a novel strategy for the future production of Vidal icewine.

Assessment of chemical composition and sensorial properties of ciders fermented with different non-Saccharomyces yeasts in pure and mixed fermentations.

This work presents the attempt to enhance the flavor complexity of cider fermented by different non-Saccharomyces species. Pichia kluyveri and Hanseniaspora vineae pure cultures were used as reference ciders. Mixed cultures between all 4 species gave 5 fermentations, where Hanseniaspora uvarum or Torulaspora quercuum were included for apple juice fermentation. Chemical composition and sensorial properties of all ciders were studied. The results indicated that the growth of P. kluyveri and H. vineae were interreacted and also affected by H. uvarum and T. quercuum. H. vineae was more capable of consuming sugar than P. kluyveri. Ciders from the single culture fermentation with P. kluyveri (Pk), as well as from mixed fermentation with P. kluyveri and H. uvarum (Pk-Hu), had high residual sugar, sugar/acid ratio, and glucose-fructose consumption ratio. Large shifts in the consumption and production of organic acids and polyphenols among all ciders were observed. The calculation of the relative odor activity value (rOAV) showed that 17 volatile compounds had an rOAV >1 in at least one sample, and acetate esters and ethyl esters were the groups with the highest number of volatile compounds of importance to the cider aroma. Among these 17 compounds, 3-methylbutyl acetate, 2-methylbutyl acetate, ethyl hexanoate, ethyl octanoate, and ß-damascenone exhibited high rOAVs in some ciders and might contribute fruity, floral, and sweet features to the cider aroma. Besides, the tropical fruity aroma from 3-methylbutyl acetate was only perceived in Pk and Pk-Hu. The partial least squares regression (PLSR) analysis revealed that acetate esters contributed positively to the roasted and cooked odor of all ciders. This is the first study evaluating simultaneous fermentation of two non-Saccharomyces yeasts to produce cider, which provides new insights into cider production.

Enhancement of Monascus pigment productivity via a simultaneous fermentation process and separation system using immobilized-cell fermentation.

A mutant Monascus purpureus strain, M183, which produced monascus pigments (MPs) at 8460 U/g via solid-state batch-fermentation, was generated using the atmospheric and room temperature plasma (ARTP) mutation system. The optimal glucose concentration (80 g/L) in traditional fermentation media that yielded the highest MPs productivity was determined. Response surface methodology (RSM) was applied to maximize MPs production using liquid-state batch-fermentation. Under optimal conditions (0.58 g/L MgSO4·7H2O, 0.02 g/L ZnSO4·7H2O, 0.02 g/L FeSO4·7H2O and 4.85 g/L NH4NO3), 207.67 U/mL of MPs were produced with 98.12% validity based on the predicted value. Extracellular MPs production increased significantly to 35.52 U/mL, compared to 14.19 U/mL of the original strain, M. purpureus LQ-6. M. purpureus spores immobilized in sodium alginate were studied. A simultaneous fermentation and separation system was established for MPs using the novel absorption resin LX300C to enhance production efficiency of extracellular MPs.

Rational synthetic combination genetic devices boosting high temperature ethanol fermentation.

The growth and production of yeast in the industrial fermentation are seriously restrained by heat stress and exacerbated by heat induced oxidative stress. In this study, a novel synthetic biology approach was developed to globally boost the viability and production ability of S. cerevisiae at high temperature through rationally designing and combing heat shock protein (HSP) and superoxide dismutase (SOD) genetic devices to ultimately synergistically alleviate both heat stress and oxidative stress. HSP and SOD from extremophiles were constructed to be different genetic devices and they were preliminary screened by heat resistant experiments and anti-oxidative experiments, respectively. Then in order to customize and further improve thermotolerance of S. cerevisiae, the HSP genetic device and SOD genetic device were rationally combined. The results show the simply assemble of the same function genetic devices to solve heat stress or oxidative stress could not enhance the thermotolerance considerably. Only S. cerevisiae with the combination genetic device (FBA1p-sod-MB4-FBA1p-shsp-HB8) solving both stress showed 250% better thermotolerance than the control and displayed further 55% enhanced cell density compared with the strains with single FBA1p-sod-MB4 or FBA1p-shsp-HB8 at 42 °C. Then the most excellent combination genetic device was introduced into lab S. cerevisiae and industrial S. cerevisiae for ethanol fermentation. The ethanol yields of the two strains were increased by 20.6% and 26.3% compared with the control under high temperature, respectively. These results indicate synergistically defensing both heat stress and oxidative stress is absolutely necessary to enhance the thermotolerance and production of S. cerevisiae.