Fermentation Cycle Research Articles

Selection and improvement of Saccharomyces cerevisiae by direct and mass mating for integrated first and second generation (1G + 2G) ethanol production

The integrated 1G + 2G ethanol production process seems to be a feasible strategy considering the Brazilian scenario, in which sugarcane molasses and bagasse are available substrates in biorefineries. However, such process imposes harsh conditions to yeasts, which must be able to simultaneously deal with several stresses from molasses and hydrolysate. In this work, improved hybrid strains of Saccharomyces cerevisiae with higher resistance for an integrated 1G + 2G ethanol production were obtained by hybridization technique. Initially, 663 wild S. cerevisiae strains were screened in microculture growth assays. Seven strains were pre-selected out of 40 based on their stress multi-tolerance traits. Subsequently, these pre-selected strains underwent evaluation in cell-recycling fed-batch fermentation conditions utilizing a medium containing molasses-bagasse hydrolysate. The two most successful strains from this evaluation were subjected to sporulation, yielding haploid cells. These haploid cells were then subjected to crossbreeding, employing direct and mass mating techniques. From the initial pool of 7 pre-selected strains, 3 hybrid strains were selected due to their enhanced resistance under the demanding conditions of cell recycling fed-batch fermentation, particularly when utilizing a higher osmotic molasses-hydrolysate medium. These hybrid strains consistently exhibited high cell viability rates and effectively accumulated elevated levels of trehalose and glycogen over the course of five fermentation cycles. Remarkably, they maintained ethanol productivity at levels comparable to their respective parental strains. The genotyping molecular identification attested the difference between the hybrids and the parental strains. The 3 hybrids selected seem to present valuable backgrounds for metabolic engineering for lignocellulosic-derived xylose fermentation.

Optimization and experimental study of variable frequency intermittent stirring strategy for anaerobic digestion based on CFD

The stirring strategy has a significant effect on the efficiency of anaerobic digestion (AD). In this study, the suspension velocity of multi-layer blade particles was characterized based on CFD simulation, and a variable frequency intermittent stirring strategy was proposed to study the stirring effect and mixing energy consumption. Firstly, the fermentation cycle was divided into three parts, and three stirring speeds and mixing times were set for each part to carry out an orthogonal combination test. Taking methane production and mixing energy consumption as optimization indexes, a net energy output equation was established to optimize the new stirring strategy. The results showed that the new variable frequency intermittent stirring strategy could effectively improve digestion efficiency. This provides a theoretical basis for setting a stirring strategy in a biogas project.

Enhanced exopolysaccharide production of Cordyceps militaris via mycelial cell immobilization on plastic composite support in repeated-batch fermentation

Repeated-batch fermentation with fungal mycelia immobilized in plastic composite support (PCS) eliminates the lag phase during fermentation and improves metabolite productivity. The strategy is implemented herein, and a novel modified PCS is developed to enhance exopolysaccharide (EPS) production from the medicinal fungus Cordyceps militaris. A modified PCS (SYE + PCS) was made by compositing polypropylene (PP) with a nutrient mixture containing soybean hull, peptone, yeast extract, and minerals (SYE+). The use of SYE + PCS has consistent cell productivity throughout the multiple fermentation cycles, which resulted in a more higher cell productivity after second batch compared to unmodified PCS. The cell grown on SYE + PCS also generates a higher yield of EPS (3.36, 6.93, and 5.72 g/L in the first, second, and third fermentation cycles, respectively) up to three-fold higher than the cell immobilized on unmodified PCS. It is also worth noting that the EPS from mycelium grown on SYE + PCS contains up to 2.3-fold higher cordycepin than those on unmodified PCS. The presence of nutrients in SYE + PCS also affects the hydrophobicity and surface roughness of the PC, improving mycelial cell adhesion. This study also provides a preliminary antioxidant activity assessment of EPS from immobilized C. militaris grown with SYE + PCS.

Optimizing the production of wheat beer by recycling the yeast biomass: perspective for cost reduction with quality maintenance

A considerable investment is being experienced worldwide in the production of artisanal beers of different styles, such as wheat-based Weizen beer. The challenge is to reduce production costs while maintaining the commercially accepted quality. Thus, the improvement of the production process involves the possibility of shortening the fermentation time by reusing the yeast biomass without affecting the yeast vitality and the quality of the end product. In this study, the fermentation parameters of a Weizen beer were measured directly from a scaled-up process in a microbrewing in order to identify the optimum point at which the process might be ended without compromising the final characteristics. The results were confirmed by laboratory studies that compared three different yeast strains in a wheat-based industrial substrate. The results led to the conclusion that the fermentation time was reduced by more than 50% from 7 to 3 days, with less variation in the concentration of the investigated metabolites. By reducing the time of fermentation, the yeast biomass was maintained active to be used up to six consecutive cycles of fermentation. All these improvements might help in reducing the cost of beer processing, which is of paramount interest to microbreweries worldwide.

Genome and fermentation analyses of Enterococcus faecalis DB-5 isolated from Japanese Mandarin orange: An assessment of potential application in lactic acid production

Enterococcus faecalis strain DB-5 is a lactic acid bacterium newly isolated from the Japanese mandarin orange (mikan). The DB-5 strain produces organic acid from various carbohydrate sources including glycerol and starch. To gain deeper insights into its potential application in lactic acid fermentation (LAF), the genome and fermentation analyses of E.faecalis DB-5 were performed. Whole genome sequencing was carried out using the DNBSEQ platform. After trimming and assembly, the total size of the assembled genome was revealed to be 3,048,630 bp, distributed into 63 contigs with an N50 value of 203,673. The genome has 37.2% GC content, 2928 coding DNA sequences, and 54 putative RNA genes. The DB-5 strain harbored two l-lactate dehydrogenases (L-LDHs), both of which conserved the catalytic domain sequences. The optical purity measurement showed that strain DB-5 is homofermentative and produced only l-lactic acid (LA), which correlated with genome-based pathway analysis. To confirm its LA productivity at high temperatures, open repeated batch fermentation was performed at 45°C using sucrose as a carbon source. The volumetric LA productivity of DB-5 was averaged at 3.66gL-1h-1 for 24h during the 3rd to 11th fermentation cycles. E.faecalis DB-5 could efficiently convert around 94% of sucrose to LA throughout the fermentation cycles at 45°C. These genomic characteristics and fermentation properties of E.faecalis DB-5 provide beneficial information for a deeper understanding of the functional properties of future high-temperature LAFs from biomass resources.

Efficient production and skincare activity evaluation of schizophyllan, a β-glucan derived from Schizophyllum commune NTU-1

Schizophyllan (SPG), a β-glucan produced by the fungus Schizophyllum commune, possesses a β-(1 → 3)-linked backbone with single β-(1 → 6)-linked glucose side chains at approximately every third residue. In this study, we screened SPG-producing strains of S. commune from different provinces in China. A candidate strain (NTU-1) with a high SPG yield was chosen, and the fermentation conditions were optimized. The optimal carbon and nitrogen sources were sucrose (40 g/L) and yeast extract (20 g/L), respectively. The optimal conditions for pH and temperature were 5.0 and 28 °C, respectively. Inclusion of 0.2 mg/L of 2,4-Dichlorophenoxyacetic acid in the medium further increased the SPG concentration. In a 5-L bioreactor, the fermentation cycle was reduced from the initial seven days to five days, and the concentration of SPG obtained was 21.3 g/L, which is the highest reported to date. In addition, we evaluated the bioactivity of the SPG prepared using strain NTU-1. The results showed that SPG had certain characteristics of anti-oxidation, anti-photoaging, and inhibition of melanin production, making it a promising reagent for skin care.

Enhanced excretion of recombinant cyclodextrin glucanotransferase and cell stability of immobilized recombinant Escherichia coli by reducing tryptone concentration

AbstractBACKGROUNDRecombinant protein excretion has become a mainstream strategy in reducing downstream processing costs. However, recombinant protein excretion is often bottlenecked by cell lysis and plasmid stability. In the present study, recombinant Escherichia coli cell immobilization was performed on a hollow fiber membrane. Tryptone concentration in the expression medium (Super Optimal Broth), which was used as a nitrogen source, was varied between 5 and 20 g L−1 to enhance the excretion of recombinant cyclodextrin glucanotransferase (CGTase), plasmid stability, and resistance of cell lysis.RESULTSThe immobilized cells with 5 g L−1 concentration of tryptone improved the plasmid stability with 119% improvement and 69% reduction of cell lysis without remarkably altering the excretion of CGTase compared with the tryptone concentration of 20 g L−1. The immobilized cells showed a 2‐fold increase in excretion of CGTase, a 45% reduction in cell lysis, and a 172% gain in plasmid stability in comparison with the free cells. Moreover, the doubling time increased to 58 and 5 h for the immobilized and free cells, respectively. The immobilized cells recorded 2301.62 U mL−1 of cumulative CGTase activity through seven fermentation cycles using the untreated membrane, marking their excellent reusability.CONCLUSIONThis new technique of recombinant protein expression utilizing an immobilized cell system under low tryptone concentration is an outstanding approach to improve recombinant CGTase excretion and plasmid stability with low cell lysis. © 2023 Society of Chemical Industry (SCI).

Directed evolution of Saccharomyces cerevisiae for low volatile acidity during winemaking under aerobic conditions

The use of yeast respiratory metabolism has been proposed as a promising approach to solve the problem of increasing ethanol content in wine, which is largely due to climate change. The use of S. cerevisiae for this purpose is mostly hampered by acetic acid overproduction generated under the necessary aerobic conditions. However, it was previously shown that a reg1 mutant, alleviated for carbon catabolite repression (CCR), showed low acetic acid production under aerobic conditions. In this work directed evolution of three wine yeast strains was performed to recover CCR-alleviated strains, expecting they will also be improved concerning volatile acidity. This was done by subculturing strains on galactose, in the presence of 2-deoxyglucose for around 140 generations. As expected, all evolved yeast populations released less acetic acid than their parental strains in grape juice, under aerobic conditions. Single clones were isolated from the evolved populations, either directly or after one cycle of aerobic fermentation. Only some clones from one of three original strains showed lower acetic acid production than their parental strain. Most clones isolated from EC1118 showed slower growth. However, even the most promising clones failed to reduce acetic acid production under aerobic conditions in bioreactors. Therefore, despite the concept of selecting low acetic acid producers by using 2-deoxyglucose as selective agent was found to be correct, especially at the population level, the recovery of strains with potential industrial utility by this experimental approach remains a challenge.

Investigation and dynamic changes of phenolic compounds during a new-type fermentation for ripened Pu-erh tea processing

At present, several novel pile-fermentation styles have been developed for ripened Pu-erh tea (RPT) processing. In this study, tea pigments, phenolic compounds, purine alkaloids and amino acids in six RPTs produced through three various fermentation styles were determined by using HPLC and amino acid analyzer to reveal chemical differences caused by fermentation style, respectively. Additionally, dynamic changes of phenolic compounds and purine alkaloids were explored during a whole new-type fermentation. Despite three various fermentation styles, these RPTs demonstrated consistent sensory and major quality components including three tea pigments, two flavonoids and three purine alkaloids. However, RPTs of the new-type fermentation had relatively higher contents of seven catechins, gallic acid, two flavonols (i.e. quercetin and kaempferol), two other flavonoids (i.e. rutin and luteolin) and 10 amino acids such as l-theanine and l-arginine. The new-type fermentation decelerated the degradation of four galloylated catechins, rutin, amino acids and soluble sugars, but contributed to the accumulations of three non-galloylated catechins, two flavonols, luteolin and gallic acid. Therefore, the new-type fermentation could be a potential style for RPT processing with a shorter fermentation cycle.

Process Evaluation and Kinetics of Recombinant Chitin Deacetylase Expression in E. coli Rosetta pLysS Cells Using a Statistical Technique.

In recent years, the greener route of the deacetylation of chitin to chitosan using the enzyme chitin deacetylase has gained importance. Enzymatically converted chitosan with emulating characteristics has a broad range of applications, particularly in the biomedical field. Several recombinant chitin deacetylases from various environmental sources have been reported, but there are no studies on process optimization for the production of these recombinant chitin deacetylases. The present study used the central composite design of response surface methodology to maximize the recombinant bacterial chitin deacetylase (BaCDA) production in E. coli Rosetta pLysS. The optimized process conditions were 0.061% glucose concentration, 1% lactose concentration, an incubation temperature of 22 °C, an agitation speed at 128 rpm, and 30 h of fermentation. At optimized conditions, the expression due to lactose induction was initiated after 16 h of fermentation. The maximum expression, biomass, and BaCDA activity were recorded 14 h post-induction. At the optimized condition, the BaCDA activity of expressed BaCDA was increased ~2.39-fold. The process optimization reduced the total fermentation cycle by 22 h and expression time by 10 h post-induction. This is the first study to report the process optimization of recombinant chitin deacetylase expression using a central composite design and its kinetic profiling. Adapting these optimal growth conditions could result in cost-effective, large-scale production of the lesser-explored moneran deacetylase, embarking on a greener route for biomedical-grade chitosan production.

Screening the Carbon Source Type in Solid-State Fermentation with Phanerochaete chrysosporium to Improve the Forage Value of Corn Straw and Rice Straw.

Poor quality straw can be made more digestible and palatable through delignification using white rot fungi as a biological treatment in SSF. The decomposition of organic matter by white rot fungi is improved when a carbon source is added. Reducing the fermentation cycle can also help retain more nutrients in straw feed. To increase rumen digestibility and nutrient utilization, corn straw and rice straw were subjected to SSF with white rot fungi (Phanerochaete chrysosporium) for 21 days. The type of carbon source (glucose, sucrose, molasses, or soluble starch) was optimized, and the nutrient composition and in vitro fermentation parameters of the fermented straw were assessed. In the fermented corn straw and rice straw supplemented with different carbon sources, the results showed a decrease in lignin content, dry matter, cellulose, and hemicellulose loss, and an increase in crude protein content after 21 days. Total volatile fatty acid and ammonium nitrogen concentrations increased significantly (p < 0.01) during in vitro fermentation. Overall, the most enhanced nutritional values for corn straw and rice straw were observed after 14 days of SSF in the groups using molasses or glucose as a carbon source.

Damage to deoxyribonucleic acid - DNA and its influence on ethanol production in industrial lines of Saccharomyces cerevisiae in relation to fermentative cycles

In industrial processes, more robust and tolerant Saccharomyces cerevisiae strains are needed, as stress conditions can affect these microorganisms and cause cellular changes. Thus, this study aimed to evaluate the genetic and physiological effect of fermentation cycles in industrial strains of S. cerevisiae. A pre-inoculum was performed by inoculating 0.10 g of Pedra-2, FT858 and Fleischmann yeasts in YPD 2% liquid medium and incubated at 30°C for 10 hours at 250 rpm. The cells were recovered and the biomass obtained was inoculated in sugarcane juice with 22°Brix at a temperature of 30°C at 250 rpm for 10 hours being conducted with cell recycling. Aliquots were taken at each cycle for genotoxicity analysis by the comet test and physiological stress by ethanol quantification. THE yeast Fleischmann showed lower tolerance to the fermentation cycle, showing greater damage to deoxyribonucleic acid (DNA). The ethanol productivity of the Fleischman strain, in the first fermentation cycle, was similar to the Pedra-2 and FT858 yeasts. However, during the cycles, there was a reduction in the content of this metabolite.

Metabolite analysis of soybean oil on promoting astaxanthin production of Phaffia rhodozyma.

Astaxanthin is a carotenoid with strong antioxidant property. In addition, it has anti-cancer, anti-tumor, anti-inflammatory and many other functions. The micro-organisms that mainly produce astaxanthin are Haematococcus pluvialis and Phaffia rhodozyma. Compared with H.pluvialis, P.rhodozyma has shorter fermentation cycle and easier to control culture conditions, but the yield of astaxanthin in P.rhodozyma is low. This article studied how to improve the astaxanthin production of P.rhodozyma. The results showed that when 10mL L-1 soybean oil was added to the culture medium, astaxanthin production increased significantly, reaching 7.35 mg L-1 , which was 1.4 times that of the control group, and lycopene and β-carotene contents also increased significantly. Through targeted metabolite analysis, the fatty acids in P.rhodozyma significantly increased under the soybean oil stimulation, especially the fatty acids closely related to the formation of astaxanthin esters, included palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1n9), linoleic acid (C18:2n6), α-linolenic acid (C18:3n3) and γ-linolenic acid (C18:3n6), thereby increasing the astaxanthin esters content. It showed that the addition of soybean oil can promote the accumulation of astaxanthin by promoting the increase of astaxanthin ester content. © 2022 Society of Chemical Industry.

Pyomelanin production via heterologous expression of 4-hydroxyphenylpyruvate dioxygenase (HPPD) and construction of HPPD inhibitor screening model

Melanin has an increasing market demand in cosmetics, food, medicine as well as aerospace due to its unique properties. Heterologous expression of 4-hydroxyphenylpyruvate dioxygenase (HPPD) from the melanin-producing strain Streptomyces fungicidicus NW-EN1 in Escherichia coli shortened the fermentation cycle of melanin. HPPD catalyzed 4-hydrophenylpyruvate (HPP) to form homologous acid (HGA) and finally form melanin. The purified melanin had the highest absorption peak at 460nm. Fourier transform infrared spectroscopy and scanning electron microscope scanning showed that the pigment had universal characteristic peaks. The presence of HGA, a predictor of pyomelanin, was identified by high-performance liquid chromatography analysis. The recombinant E.coli produced 804.4±5.9mg/L pyomelanin within 48h. Metal ions had a great influence on the production of pyomelanin. Pyomelanin was stable in response to light intensity and had a protective effect against bacteria under UV irradiation. Meanwhile, we utilized the chromogenic effect after whole-cell catalysis to reflect the inhibition of the HPPD inhibitors (mesotrione and isoxaflutole) on HPPD by observing the color change. As a rapid method to test the action of inhibitors, this method is expected to be useful for the development of HPPD-inhibiting herbicides.

Effects of Jiuqu inoculating Rhizopus oryzae Q303 and Saccharomyces cerevisiae on chemical components and microbiota during black glutinous rice wine fermentation

In this study, the physicochemical properties, metabolites, sensory characteristics, and microbiota of black glutinous rice wine (BGRW) during traditional fermentation (TF) and inoculated fermentation (IF) were investigated, and their correlation relationships were revealed. Results indicated that IF promoted total sugar utilization and ethanol and esters synthesis in BGRW. Altogether 67 reliable metabolites were identified, 36 of which were labeled components, primarily esters (OAV > 1). Meanwhile, the concentrations of ethanol, esters, and total flavor on the 11th day of IF were close to those on the 22nd day of TF. Combined with the sensory description indicated that the IF could improve the quality of BGRW. Besides, TF contains vast unfavorable microorganisms, such as Chaetomium, Penicillium, Stachybotrys, and Trichocladium. Therefore, IF can accelerate the fermentation and enhance the flavor of BGRW, and inhibit the growth of pathogenic and spoilage organisms. Finally, Pearson's correlation analysis demonstrated that Saccharomyces, Rhizopus, Lactobacillus, Aspergillus, and Bacillus were the pivotal functional microorganisms in BGRW. The study provided scientific and effective basis for improving the quality, shortening the fermentation cycle, and controlling pathogenic and spoilage organisms of BGRW.

Efficiency of magnetic immobilization for recombinant Pichia pastoris cells harvesting over consecutive production cycles

ABSTRACT In the current experiment, P. pastoris cells were selected as promising candidate for production of recombinant proteins and were magnetically immobilized by 3-aminopropyltriethoxysilane-coated magnetite nanoparticles (APTES@Fe3O4). The ability of immobilized cells for the production of recombinant human serum albumin (HSA) and the potency of magnetic immobilization for yeast cells recycling was evaluated over three successive batch fermentation cycles. Interestingly, magnetic immobilization resulted in 1.4-fold increase in the HAS production. The productivity was increased from 0.7 mg/mL up to one mg/mL. But, after each production cycle a significant (more than two-fold) reduction in the productivity was recorded in free and immobilized cells. Over consecutive production cycles, efficiency of magnetic immobilization was also reduced. After three production cycles, immobilization efficiency was reduced from 80% down to 64% that was due to appearance of magnetic insensitive cells. Field emission scanning electron microscopy (FESEM) analyses approved that magnetic insensitive cells are nanoparticles free cells. These free cells can be resulted from cell proliferation and detachment of nanoparticles from cells surface. These findings can be considered for development of an efficient immobilization technique to harvest yeast cells over consecutive production cycles.

Effects of multiple cycles of sorghum starch gelatinization and fermentation on production of Chinese strong flavor Baijiu

The property changes of sorghum starch during multiple cycles of gelatinization and fermentation were studied. This study simulated the gelatinization and fermentation process of raw starch in strong-flavor Baijiu production (Sorghum as raw material, the distillers' grains as a control). The results showed that the starch content of the same batch of sorghum after five cycles of gelatinization and fermentation was 9.98% (Cannot continue to be used for fermentation), and about 60% of the starch was consumed in the first three cycles of gelatinization and fermentation. The gel properties of sorghum starch gradually decreased during fermentation but slightly increased after gelatinization. After five cycles of gelatinization and fermentation, the sorghum starch has a uniform size and a thin and small fragment structure. At the same time, sorghum starch does not form new groups. The recrystallization of starch caused by multiple cycles of gelatinization and fermentation increased the onset gelatinization temperature (from 61.6 to 114.4 °C), peak gelatinization temperature (from 78.5 to 139.5 °C), and gelatinization enthalpy (from 7.980 to 17.121 J/g) of sorghum starch. The crystalline structure of sorghum starch changed from the initial type A to type A + V, type A + B + V, and finally to type B.

Fed-Batch Fermentation of Saccharomyces pastorianus with High Ribonucleic Acid Yield.

(1) Background: The degradation products of ribonucleic acid (RNA)are widely used in the food and pharmaceutical industry for their flavoring and nutritional enhancement functions. Yeast is the main source for commercial RNA production, and an efficient strain is the key to reducing production costs; (2) Methods: A mutant Saccharomyces pastorianus G03H8 with a high RNA yield was developed via ARTP mutagenesis and fed-batch fermentation was applied to optimize production capacity. Genome sequencing analysis was used to reveal the underlying mechanism of higher RNA production genetic differences in the preferred mutant; (3) Results: Compared with the highest RNA content of the mutant strain, G03H8 increased by 40% compared with the parental strain G03 after response surface model optimization. Meanwhile, in fed-batch fermentation, G03H8′s dry cell weight (DCW) reached 60.58 g/L in 5 L fermenter by molasses flowing and RNA production reached up to 3.58 g/L. Genome sequencing showed that the ribosome biogenesis, yeast meiosis, RNA transport, and longevity regulating pathway were closely related to the metabolism of high RNA production; (4) Conclusion: S. pastorianus G03H8 was developed for RNA production and had the potential to greatly reduce the cost of RNA production and shorten the fermentation cycle. This work lays the foundation for efficient RNA content using S. pastorianus.

Metabolome analysis of the response and tolerance mechanisms of Saccharomyces cerevisiae to formic acid stress.

Various inhibitors are produced during the hydrolysis of lignocellulosic biomass that can interfere with the growth of yeast cells and the production of bioethanol. Formic acid is a common weak acid inhibitor present in lignocellulosic hydrolysate that has toxic effects on yeast cells. However, the mechanism of the response of Saccharomyces cerevisiae to formic acid is not fully understood. In this study, liquid chromatography-mass spectrometry (LC-MS) was used to investigate the effects of formic acid treatment on cell metabolites of S. cerevisiae. Treatment with different concentrations of formic acid significantly inhibited the growth of yeast cells, reduced the yield of ethanol, prolonged the cell fermentation cycle, and increased the content of malondialdehyde. Principal component analysis and orthogonal partial least squares discriminant analysis showed that 55 metabolites were significantly altered in S. cerevisiae after formic acid treatment. The metabolic relevance of these compounds in the response of S. cerevisiae to formic acid stress was investigated. Formic acid can cause oxidative stress, inhibit protein synthesis, and damage DNA in S. cerevisiae, and these are possible reasons for the inhibition of S. cerevisiae cell growth. In addition, the levels of several aromatic amino acids identified in the cells of formic acid-treated yeast were increased; the biosynthesis of nucleotides was slowed, and energy consumption was reduced. These mechanisms may help to improve the tolerance of yeast cells to formic acid. The results described herein highlight our current understanding of the molecular mechanism of the response of S. cerevisiae to formic acid. The study will provide a theoretical basis for research on the tolerance mechanisms of S. cerevisiae.

Development and Analysis of an Intensified Batch-Fed Wine Fermentation Process

White wine fermentations are typically performed in an entirely batchwise manner, with yeast nutrients only added at the beginning of fermentation. This leads to slow (2+ weeks) fermentation cycle times, with large capital expenditures required to increase winery processing capacity. Prior attempts to speed fermentations via increasing temperature have resulted in unpalatable wine, and continuous fermentation processing is uneconomical and impractical in the winery setting. In this work, we measured yeast nutrient consumption as a function of fermentation progression at the 300 mL scale, and from this derived an equation to optimize yeast nutrient concentration as a function of fermentation progression. These findings were applied at the pilot scale in 150 L fermentors, which resulted in a 60% cycle time reduction versus “best practices” control fermentations. The resultant wines were compared via GC-MS as well as by a trained sensory panel. Organoleptic analysis found statistically significant, but overall, small differences in sensory characteristics between the control and process intensified wines. This intensified fermentation process shows great promise for fermented beverage producers wishing to maximize equipment utilization and debottleneck wineries or other beverage fermentation facilities.