Industrial Fermentation Processes Research Articles

Engineering Implementation of the Acosta Fermentation Method to Obtain Cuban Schnapps with Reduced Concentrations of Higher Alcohols

The Acosta method involves rewiring the yeast metabolic pathway to enhance biomass production and prevent a significant increase in higher alcohols during the late stages of fermentation. This study aimed to assess fermentation conditions to achieve Cuban schnapps with reduced concentrations of higher alcohols and replicate the process on an industrial scale. To achieve this, the quality of final sugarcane molasses for fermentation by Saccharomyces cerevisiae (S. cerevisiae) yeast was evaluated. Industrial pre-fermentation and fermentation processes were successfully conducted, followed by laboratory-scale fermentation using the Acosta fermentation method to determine crucial parameters for industrial implementation. Operational parameters for fermentation were identified from the following results: 13.5 °Brix seeding, metabolic pathway inversion of S. cerevisiae at 16 h, and an air concentration of 0.1 m3/min. The resulting Cuban schnapps obtained using this method exhibited a concentration of higher alcohols of 132.5 mg/L, a value that is within the standard parameters, showing a positive impact of this fermentation method on the quality of the schnapps. Scaling up this method to an industrial level, in addition to offering higher quality products and being an economically viable alternative, also stands out for its sustainable and environmentally friendly aspect, and results in higher production of yeast biomass as a byproduct, which can be used for various purposes, such as animal feed. This method constitutes an important update to the schnapps production process as a technological improvement that respects sustainable production trends and the characteristics of the final product.

Research on the Microbial Community Structure and Evolutionary Changes During the Industrial Fermentation Process of Cigar Tobacco Leaves

Cigar tobacco leaves host a diverse bacterial community on their surfaces, playing a vital role in the fermentation process. This study aimed to investigate and analyze the bacterial community dynamics during the artificial fermentation of cigar tobacco leaves, recognizing the advantages of artificial over natural fermentation. The research focused on elucidating the structural composition of the bacterial community on the leaves, exploring their metabolic activities, understanding how they influence the characteristics of the leaves during artificial fermentation, and identifying potential metabolic pathways. Significant shifts in the microbial community were observed post-fermentation, with a marked increase in diversity. Notably, genera like Oceanobacillus, Terribacillus, and Corynebacterium exhibited substantial growth during the mid-phase of fermentation, suggesting their importance in the process. The insights derived from this study shed light on the artificial fermentation process of cigar tobacco leaves, offering opportunities for enhancing safety and quality.

Achieving robust synthetic tolerance in industrial E. coli through negative auto-regulation of a DsrA-Hfq module

In industrial fermentation processes, microorganisms often encounter acid stress, which significantly impact their productivity. This study focused on the acid-resistant module composed of small RNA (sRNA) DsrA and the sRNA chaperone Hfq. Our previous study had shown that this module improved the cell growth of Escherichia coli MG1655 at low pH, but failed to obtain this desired phenotype in industrial strains. Here, we performed a quantitative analysis of DsrA-Hfq module to determine the optimal expression mode. We then assessed the potential of the CymR-based negative auto-regulation (NAR) circuit for industrial application, under different media, strains and pH levels. Growth assay at pH 4.5 revealed that NAR-05D04H circuit was the best acid-resistant circuit to improve the cell growth of E. coli MG1655. This circuit was robust and worked well in the industrial lysine-producing strain E. coli SCEcL3 at a starting pH of 6.8 and without pH control, resulting in a 250 % increase in lysine titer and comparable biomass in shaking flask fermentation compared to the parent strain. This study showed the practical application of NAR circuit in regulating DsrA-Hfq module, effectively and robustly improving the acid tolerance of industrial strains, which provides a new approach for breeding industrial strains with tolerance phenotype.

Investigating the Role of β-Disodium Glycerophosphate and Urea in Promoting Growth of Streptococcus thermophilus from Omics-Integrated Genome-Scale Models.

This study investigates the impact of urea and β-GP on the growth of Streptococcus thermophilus S-3, a bacterium commonly used in industrial fermentation processes. Through a series of growth experiments, transcriptome, metabolome, and omics-based analyses, the research demonstrates that both urea and β-GP can enhance the biomass of S. thermophilus, with urea showing a more significant effect. The optimal urea concentration for growth was determined to be 3 g/L in M17 medium. The study also highlights the metabolic pathways influenced by urea and β-GP, particularly the galactose metabolism pathway, which is crucial for cell growth when lactose is the substrate. The integration of omics data into the genome-scale metabolic model of S. thermophilus, iCH502, allowed for a more accurate prediction of metabolic fluxes and growth rates. The study concludes that urea can serve as a viable substitute for β-GP in the cultivation of S. thermophilus, offering potential cost and efficiency benefits in industrial fermentation processes. The findings are supported by validation experiments with 11 additional strains of S. thermophilus, which showed increased biomass in UM17 medium.

Circular economyeast: Saccharomyces cerevisiae as a sustainable source of glucans and its safety for skincare application

Glucans, a polysaccharide naturally present in the yeast cell wall that can be obtained from side streams generated during the fermentation process, have gained increasing attention for their potential as a skin ingredient. Therefore, this study focused on the extraction method to isolate and purify water-insoluble glucans from two different Saccharomyces cerevisiae strains: an engineered strain obtained from spent yeast in an industrial fermentation process and a wild strain produced through lab-scale fermentation. Two water-insoluble extracts with a high glucose content (> 90 %) were achieved and further subjected to a chemical modification using carboxymethylation to improve their water solubility. All the glucans' extracts, water-insoluble and carboxymethylated, were structurally and chemically characterized, showing almost no differences between both yeast-type strains. To ensure their safety for skin application, a broad safety assessment was undertaken, and no cytotoxic effect, immunomodulatory capacity (IL-6 and IL-8 regulation), genotoxicity, skin sensitization, and impact on the skin microbiota were observed. These findings highlight the potential of glucans derived from spent yeast as a sustainable and safe ingredient for cosmetic and skincare formulations, contributing to the sustainability and circular economy.

Analyzing microbial community and volatile compound profiles in the fermentation of cigar tobacco leaves

Variations in industrial fermentation techniques have a significant impact on the fermentation of cigar tobacco leaves (CTLs), consequently influencing the aromatic attributes of the resulting cigars. The entire fermentation process of CTLs can be categorized into three distinct phases: phase 1 (CTLs prior to moisture regain), phase 2 (CTLs post-moisture regain and pile fermentation), and phase 3 (CTLs after fermentation and drying). These phases were determined based on the dynamic changes in microbial community diversity. During phase 2, there was a rapid increase in moisture and total acid content, which facilitated the proliferation of Aerococcus, a bacterial genus capable of utilizing reducing sugars, malic acid, and citric acid present in tobacco leaves. In contrast, fungal microorganisms exhibited a relatively stable response to changes in moisture and total acid, with Aspergillus, Alternaria, and Cladosporium being the dominant fungal groups throughout the fermentation stages. Bacterial genera were found to be more closely associated with variations in volatile compounds during fermentation compared to fungal microorganisms. This association ultimately resulted in higher levels of aroma components in CTLs, thereby improving the overall quality of the cigars. These findings reinforce the significance of industrial fermentation in shaping CTL quality and provide valuable insights for future efforts in the artificial regulation of secondary fermentation in CTLs.Key points• Industrial fermentation processes impact CTLs microbial communities.• Moisture and total acid content influence microbial community succession in fermentation.• Bacterial microorganisms strongly influence CTLs’ aldehyde and ketone flavors over fungi.

Microbiological and Physicochemical Dynamics in Traditional and Industrial Fermentation Processes of Koumiss

Koumiss, a traditional fermented beverage made from mare’s milk, is typically consumed by nomads. Industrialized production of koumiss has been increasingly applied recently due to the increased demand for the beverage and awareness of its potential health benefits. However, it is unknown whether industrial koumiss is comparable to the traditional koumiss in terms of quality. In this study, we compared the microbiological and physicochemical properties in the industrial and traditional koumiss fermentation processes synchronously using culture-dependent and culture-independent approaches. Although Lactobacillus and Kazachstania species were similarly dominant in the bacterial and fungal communities, respectively, in both processes, the microbial counts and diversity in the traditional koumiss were significantly higher than those in the industrial koumiss. Furthermore, the traditional koumiss fermentation consumed more lactose, produced more flavor substances including acetic acid, lactic acid, ethanol, and free amino acids, and reached a lower pH value at the final stage. The physicochemical characters of traditional koumiss were mainly associated with Lactobacillus and Kazachstania species, which, in turn, were positively correlated with each other but negatively correlated with other non-dominant microbes. The starter was the major source of the microbial community of industrial koumiss, whereas both the starter and environment were the major sources of traditional koumiss. Random forest analysis recognized 11 significantly important genera as microbial indicators to distinguish industrial from traditional koumiss. Overall, this study shows that the microbial and physicochemical dynamics during the traditional and industrial fermentation of koumiss differ significantly, and the results obtained are valuable for improving the quality of industrial koumiss.

A Physiogenomic Study of the Tolerance of Saccharomyces cerevisiae to Isoamyl Alcohol

Isoamyl alcohol is a clear, unpleasantly odorous, colorless liquid of higher alcohol that emits a fruity aroma when heavily diluted. It has received much attention in recent years as a new fuel with a high energy density. Isoamyl alcohol can be produced industrially by microbial fermentation. Still, its toxicity to host cells has limited its potential for industrial production, and the molecular mechanism of its toxic effects has not yet been elucidated. In this study, RNA-Seq technology was used to analyze the transcripts of Saccharomyces cerevisiae under normal conditions and in the presence of isoamyl alcohol (0.5 g/L and 2.5 g/L). The results showed that the expression of the cell wall (CCW12, BGL2, NCW2 and SUN4), cell membrane (ELO1, ERG2, FAA1, and OPI3), translation and other structural genes were significantly down-regulated. The expression of genes related to ATP biosynthesis, NADPH biosynthesis (ZWF1), and metal ion transport (PMC1) proteins were up-regulated. Strains with key genes knocked out were cultured without isoamyl alcohol. Combined results suggested that isoamyl alcohol may affect cell wall stability and cell membrane fluidity, and the expression of genes related to ion homeostasis and energy production may play a protective role against isoamyl alcohol stress. By maintaining cell wall stability/membrane fluidity under isoamyl alcohol pressure, improving certain ion homeostasis, and generating energy/NADPH, it is possible to overcome the toxicity of isoamyl alcohol in industrial fermentation processes to a certain extent.

Selected cachaça yeast strains share a genomic profile related to traits relevant to industrial fermentation processes.

The isolation and selection of yeast strains to improve the quality of the cachaça-Brazilian Spirit-have been studied in our research group. Our strategy considers Saccharomyces cerevisiae as the predominant species involved in sugarcane juice fermentation and the presence of different stressors (osmolarity, temperature, ethanol content, and competition with other microorganisms). It also considers producing balanced concentrations of volatile compounds (higher alcohols and acetate and/or ethyl esters), flocculation capacity, and ethanol production. Since the genetic bases behind these traits of interest are not fully established, the whole genome sequencing of 11 different Saccharomyces cerevisiae strains isolated and selected from different places was analyzed to identify the presence of a specific genetic variation common to cachaça yeast strains. We have identified 20,128 single-nucleotide variants shared by all genomes. Of these shared variants, 37 were new variants (being six missenses), and 4,451 were identified as missenses. We performed a detailed functional annotation (using enrichment analysis, protein-protein interaction network analysis, and database and in-depth literature searches) of these new and missense variants. Many genes carrying these variations were involved in the phenotypes of flocculation, tolerance to fermentative stresses, and production of volatile compounds and ethanol. These results demonstrate the existence of a genetic profile shared by the 11 strains under study that could be associated with the applied selective strategy. Thus, this study points out genes and variants that may be used as molecular markers for selecting strains well suited to the fermentation process, including genetic improvement by genome editing, ultimately producing high-quality beverages and adding value.IMPORTANCEThis work demonstrates the existence of new genetic markers related to different phenotypes used to select yeast strains and mutations in genes directly involved in producing flavoring compounds and ethanol, and others related to flocculation and stress resistance.

Combining bio- and chemo-catalysis for cyclic carbonate synthesis at very low pressure: Efficient “online” utilization of CO2 obtained directly from the fermentation of sugars

A very low-pressure set-up for the synthesis of cyclic carbonates and polycarbonates from epoxides/CO2 has been constructed using our previously reported Ga-catalyst. This set-up is based on the direct use of unpurified CO2 obtained from the fermentation of sugars, which exemplifies a potentially important advancement for the future application of this widely explored CO2 utilization reaction. Using this approach, it has been possible to produce cyclic carbonates with a high bio-derived atom content when starting from bio-based epoxides, such as, eugenol oxide. Use of alternative sources of sugar beyond sucrose have also been exemplified (grapes), highlighting the potential for direct coupling to important industrial fermentation processes which emit large amounts of CO2. Finally, use of 13C6-glucose as sugar-source has allowed for the preparation of 13C-containing cyclic carbonates, with the bis-cyclic carbonates being readily converted through reaction with a diamine to form 13C-labelled polymers (13C-Non-Isocyanate PolyUrethanes; 13C-NIPU’s).

Dynamic nonlinear soft sensor modelling method using linear slow feature analysis and least squares support vector regression for batch processes

AbstractData‐driven soft sensor models have been extensively utilized in industrial processes. Batch processes are usually employed to manufacture low‐volume and high value‐added products in chemical, materials, and pharmaceutical industries. The most distinctive features of batch process lie in nonlinear, repetition, and slow time varying characteristics. In this paper, a data‐driven soft sensor modelling method based on linear slow feature analysis (LSFA) and least squares support vector regression (LSSVR) is proposed. In this method, LSFA was used to effectively capture the driving force behind the data features that change as slowly as possible. Then, a LSSVR model was constructed between the extracted slow feature variables and quality variables. Finally, a numerical example, industrial penicillin fermentation processes, and cobalt oxalate synthesis process were utilized to confirm the prediction accuracy and model reliability of the proposed approach.

Biotechnological production of ectoine: current status and prospects.

Ectoine is an important natural secondary metabolite in halophilic microorganisms. It protects cells against environmental stressors, such as salinity, freezing, drying, and high temperatures. Ectoine is widely used in medical, cosmetic, and other industries. Due to the commercial market demand of ectoine, halophilic microorganisms are the primary method for producing ectoine, which is produced using the industrial fermentation process "bacterial milking." The method has some limitations, such as the high salt concentration fermentation, which is highly corrosive to the equipment, and this also increases the difficulty of downstream purification and causes high production costs. The ectoine synthesis gene cluster has been successfully heterologously expressed in industrial microorganisms, and the yield of ectoine was significantly increased and the cost was reduced. This review aims to summarize and update microbial production of ectoine using different microorganisms, environments, and metabolic engineering and fermentation strategies and provides important reference for the development and application of ectoine.

Soft sensor based on multi‐phase stacking ensemble model with self‐selected primary learner for batch processes

AbstractIn batch processes, existing soft sensing methodologies encounter substantial challenges when confronted with nonlinearity and multi‐phase issues. In response to these challenges, an innovative soft sensing framework known as the multi‐phase stacking ensemble model with self‐selected primary learner is proposed. The main innovation of this framework lies in the solution to the primary learner selection issue within the stacking model. To commence, the batch process is divided into multiple phases employing a Gaussian mixture model, thereby establishing local ensemble models for each phase. Subsequently, the iterative self‐selection of primary learners strategy is proposed, which iteratively selects suitable primary learners for these models, optimizing their combination of primary learners for each local model. This primary learner selection strategy effectively enhances the accuracy of predictions in the stacking ensemble model. To further enhance the performance, Bayesian optimization is utilized to tune the hyperparameters of each local ensemble model. This step guarantees optimal performance of the model across diverse phases. Extensive simulation experiments are conducted on an industrial penicillin fermentation process to validate the effectiveness of the proposed framework. According to the findings, the model demonstrated superior performance compared to existing single‐learner soft sensing methods and commonly utilized ensemble‐based soft sensing methods in terms of both R2 score (0.97584) and RMSE (0.0513). Overall, this framework offers a novel approach for selecting primary learners in stacking ensemble models and enhancing the predictive performance in batch processes for soft sensing.

Carbon and nitrogen sources consumption by ale and lager yeast strains: a comparative study during fermentation.

The rapid and efficient consumption of carbon and nitrogen sources by brewer's yeast is critical for the fermentation process in the brewing industry. The comparison of the growth characterizations of typical ale and lager yeast, as well as their consumption preference to carbon and nitrogen sources were investigated in this study. Results showed that the ale strain grew faster and had a more extended stationary phase than the lager strain. However, the lager strain was more tolerant to the stressful environment in the later stage of fermentation. Meanwhile, the ale and lager yeast strains possessed varying preferences for metabolizing the specific fermentable sugar or free amino acid involved in the wort medium. The lager strain had a strong capacity to synthesize the extracellular invertase required for hydrolyzing sucrose as well as a strong capability to metabolize glucose and fructose. Furthermore, the lager strain had an advantage in consuming Lys, Arg, Val, and Phe, whereas the ale strain had a higher assimilation rate in consuming Tyr. These findings provide valuable insights into selecting the appropriate brewer's yeast strain based on the wort components for the industrial fermentation process. KEY POINTS: • The lager strain is more tolerant to the stressful environment. • The lager strain has the great capability to synthesize the extracellular invertase. • The assimilation efficiency of free amino acid varies between ale and lager.

Application of Enzyme Preparation as a Coadjuvant in the Control of Foam Formation in the Bioethanol Industrial Fermentation Process

Application of Enzyme Preparation as a Coadjuvant in the Control of Foam Formation in the Bioethanol Industrial Fermentation Process

Solvent-Producing Clostridia Revisited.

The review provides an overview of the current status of the solvent-producing clostridia. The origin and development of industrial clostridial species, as well as the history of the industrial Acetone Butanol Ethanol fermentation process, is reexamined, and the recent resurgence of interest in the production of biobutanol is reviewed. Over 300 fully sequenced genomes for solvent-producing and closely related clostridial species are currently available in public databases. These include 270 genomes sourced from the David Jones culture collection. These genomes were allocated arbitrary DJ codes, and a conversion table to identify the species and strains has now been provided. The expanded genomic database facilitated new comparative genomic and phylogenetic analysis. A synopsis of the common features, molecular taxonomy, and phylogeny of solvent-producing clostridia and the application of comparative phylogenomics are evaluated. A survey and analysis of resident prophages in solvent-producing clostridia are discussed, and the discovery, occurrence, and role of novel R-type tailocins are reported. Prophage genomes with R-type tailocin-like features were detected in all 12 species investigated. The widespread occurrence of tailocins in Gram-negative species is well documented; this survey has indicated that they may also be widespread in clostridia.

Insights into the mechanism of mycelium transformation of Streptomyces toxytricini into pellet.

Formation of the mycelial pellet in submerged cultivation of Streptomycetes is unwanted in industrial fermentation processes as it imposes mass transfer limitations, changes in the rheology of a medium, and affects the production of secondary metabolites. Though detailed information is not available about the factors involved in regulating mycelial morphology, it is studied that culture conditions and the genetic information of strain play a crucial role. Moreover, the proteomic study has revealed the involvement of low molecular weight proteins such as; DivIVA, FilP, ParA, Scy, and SsgA proteins in apical growth and branching of hyphae, which results in the establishment of the mycelial network. The present study proposes the mechanism of pellet formation of Streptomyces toxytricini (NRRL B-5426) with the help of microscopic and proteomic analysis. The microscopic analysis revealed that growing hyphae contain a bud-like structure behind the apical tip, which follows a certain organized path of growth and branching, which was further converted into the pellet when shake flask to the shake flask inoculation was performed. Proteomic analysis revealed the production of low molecular weight proteins ranging between 20 and 95kDa, which are involved in apical growth and hyphae branching and can possibly participate in the regulation of pellet morphology.

Effects of adding cocoa fermentation medium on cigar leaves in agricultural fermentation stage.

Background and Objective: With the development of the world economy and the integration of cultures, the Chinese cigar market has shown a significant upward trend. However, high-quality cigar leaves are mostly produced in Dominica, Cuba, Nicaragua and other places. In contrast, Chinese cigar leaves have problems such as insufficient aroma, which has become one of the main factors restricting the development of Chinese cigars. Adding medium to ferment is a traditional method in the cigar industry. At present, it mostly relies on manual experience, and lacks systematic and scientific research. At the same time, the addition of medium fermentation is mainly concentrated in the industrial fermentation process, and has not yet begun to be applied in the agricultural fermentation process. In this study, the medium was added to the agricultural fermentation process for the first time to explore the possibility of the application. The effects of adding cocoa medium to ferment on the chemical composition, sensory quality and surface microbial diversity of eggplant core cigar leaves were investigated.wrapper. Method: With Dexue 7' as the experimental material, the changes of main chemical components of wrapper fermented with water and cocoa medium were determined, and microbial community structure on the surface and relative abundance of cigar leaves at different turning periods were analyzed, and the functional genes were predicted. The results of the study were as follows: 1) The results of sensory evaluation showed that the addition of cocoa medium could highlight the aroma of bean, cocoa and coffee, improve the sweetness and fluency and the combustibility of cigar leaves. 2) The addition of cocoa medium increased the contents of proline and malic acid which were positively correlated with sensory quality, and decreased the contents of citric acid, linoleic acid, basic amino acids and aromatic amino acids which were negatively correlated with sensory quality. 3) The addition of cocoa medium increased the total amount of aroma components in cigar leaves, especially carotenoid degradation products, and changed the structural composition of some aroma substances in wrappercigar leaves. 4) The similarity of species composition between the water-added group and the cocoa-added group was higher, but the dominant microorganisms were more concentrated. Staphylococcus and Arthrobacter maintained a high relative abundance throughout the fermentation process, which may be the key microorganisms in the agricultural fermentation stage. 5) The addition of cocoa medium increased the expression abundance of related functional genes in cigar leaves, accelerated the fermentation process of cigar leaves, and bacteria played a major role in the fermentation process. Conclusion: Adding cocoa medium in the agricultural fermentation stage, the changes of bacterial community and dominant flora on the surface of cigar leaves are the main factors affecting their internal chemical components, and the addition of media has a positive effect on tobacco fermentation.

Single and multi-objective dynamic optimization study of an industrial scale fed batch reactor

Abstract The present work focuses on obtaining optimal operational policies of an industrial scale penicillin fed-batch fermentation process using dynamic optimization (DO). The three process objectives considered in this work include, maximization of total penicillin formed (J 1), minimization of total biomass formed (J 2), and minimization of fed-batch operation time (J 3). The control variables for the optimization study include, the feed flowrates of sugar, soyabean oil, and phenylacetic. We perform a single-objective optimization study with J 1 as the objective function and showcased the importance of DO. Consequently, we solve three multi-objective optimization (MOO) problems corresponding to all the pairwise combinations of the aforementioned three objectives. Outcomes of MOO problems are presented in terms of several non-dominated Pareto-optimal solutions. Furthermore, descriptive analysis of three representative Pareto points for all the MOO problems is presented, which highlights the importance of MOO study of the penicillin bio-reactor.

Attention-based LSTM Block Model Framework based on static and dynamic variables for modeling fuel ethanol fermentation process

In the industrial fuel ethanol process, the initial feed conditions, which are static variables, and each control operation, which is dynamic variable and is changing during the producing process, have an impact on the concentration of ethanol out of the tank. Developing the accurate concentration model of ethanol out of the tank with the static variables and as early as possible dynamic variables is a challenging work and is useful in analyzing and optimizing the production process. Given that the ethanol fermentation process is multiphase and dynamic, a block concentration model of ethanol out of the tank based on multilayer perceptron (MLP) and long short-term memory (LSTM) is proposed to deal with the coexistence state of the static variables and the dynamic variables. A Channel-Squeeze-and-Excitation (CSE) module is constructed to solve the problem of high-dimensional input and low-dimensional output caused by the block model. The proposed model is applied for the industrial ethanol fermentation process. The results show that CSE-MLSTM is significantly different from other prediction models and improves the prediction accuracy. Temperature control experiments and ablation experiments effectively verify the reliability and effectiveness of CSE-MLSTM.