Kinetic Modeling of Saccharomyces cerevisiae Central Carbon Metabolism: Achievements, Limitations, and Opportunities

Central carbon metabolism comprises the metabolic pathways in the cell that process nutrients into energy, building blocks and byproducts. To unravel the regulation of this network upon glucose perturbation, several metabolic models have been developed for the microorganism Saccharomyces cerevisiae. These dynamic representations have focused on glycolysis and answered multiple research questions, but no commonly applicable model has been presented. This review systematically evaluates the literature to describe the current advances, limitations, and opportunities. Different kinetic models have unraveled key kinetic glycolytic mechanisms. Nevertheless, some uncertainties regarding model topology and parameter values still limit the application to specific cases. Progressive improvements in experimental measurement technologies as well as advances in computational tools create new opportunities to further extend the model scale. Notably, models need to be made more complex to consider the multiple layers of glycolytic regulation and external physiological variables regulating the bioprocess, opening new possibilities for extrapolation and validation. Finally, the onset of new data representative of individual cells will cause these models to evolve from depicting an average cell in an industrial fermenter, to characterizing the heterogeneity of the population, opening new and unseen possibilities for industrial fermentation improvement.

Preparation, physicochemical and microbiological evaluations of italian type salami fermented with kefir

Industrialized meat product, known as salami, is composed of some types of meat, such as pork, beef or pork and beef together, added ingredients and bacon. This research aimed to elaborate Italian salami using the fermentation process with standard starter bacteria, such as control, water and milk kefir, and evaluate the physicochemical and microbiological characteristics of the final product. The use of kefir grains aimed to analyze the possibility for their use in the production of salami as starter cultures from their symbiotic association of yeasts, acid-lactic bacteria, acetic bacteria and also their probiotic and antimicrobial action. The microbiological and physicochemical composition indicates that kefir is a product with probiotic characteristics, because it has in its composition living microorganisms capable of improving the intestinal microbial balance producing beneficial effects on the health of the individual who consumes it. At the end of the analyses, it was observed that the salami based on standard starter bacteria culture presented greater diversity of fungi, and the fungus Exophiala spp was present in the 3 types of salamis. The samples showed absence of pathogenic bacteria such as Salmonella ssp, Shigella ssp and E.coli. There was little variation in the physical-chemical characteristics of the salamis evaluated. It was concluded that kefir can replace industrial fermentation culture without affecting the microbiological and physicochemical pattern of salami.

Practical considerations for delivering on the sustainability promise of fermentation-based biomanufacturing

Increasingly, bio-based products made via sugar-powered microbial cell factories and industrial fermentation are reaching the market and presenting themselves as sustainable alternatives to fossil and animal-based products. The sustainability potential of biotechnology, however, has been shown to come with trade-offs and cannot be taken for granted. Shared environmental impact hotspots have been identified across industrial fermentation-based products, including biomass production, energy consumption, and end-of-life fate. Based on both these patterns and our direct experience in preparing for the commercial-scale production of Brewed Protein™, we outline practical considerations for improving the sustainability performance of bio-based products made via industrial fermentation.

A Wild Yeast Laboratory Activity: From Isolation to Brewing

Microbial fermentation is a common form of metabolism that has been exploited by humans to great benefit. Industrial fermentation currently produces a myriad of products ranging from biofuels to pharmaceuticals.

Phytosterols and Novel Triterpenes Recovered from Industrial Fermentation Coproducts Exert In Vitro Anti-Inflammatory Activity in Macrophages

The unstoppable growth of human population that occurs in parallel with all manufacturing activities leads to a relentless increase in the demand for resources, cultivation land, and energy. In response, currently, there is significant interest in developing strategies to optimize any available resources and their biowaste. While solutions initially focused on recovering biomolecules with applications in food, energy, or materials, the feasibility of synthetic biology in this field has been demonstrated in recent years. For instance, it is possible to genetically modify Saccharomyces cerevisiae to produce terpenes for commercial applications (i.e., against malaria or as biodiesel). But the production process, similar to any industrial activity, generates biowastes containing promising biomolecules (from fermentation) that if recovered may have applications in different areas. To test this hypothesis, in the present study, the lipid composition of by-products from the industrial production of β-farnesene by genetically modified Saccharomyces cerevisiae are studied to identify potentially bioactive compounds, their recovery, and finally, their stability and in vitro bioactivity. The assayed biowaste showed the presence of triterpenes, phytosterols, and 1-octacosanol which were recovered through molecular distillation into a single fraction. During the assayed stability test, compositional modifications were observed, mainly for the phytosterols and 1-octacosanol, probably due to oxidative reactions. However, such changes did not affect the in vitro bioactivity in macrophages, where it was found that the obtained fraction decreased the production of TNF-α and IL-6 in lipopolysaccharide (LPS)-induced inflammation.

Scale-up of Aerated Industrial Multistage Rushton Impeller Bioreactors with Complex Rheology

The power input and gas-liquid mass transfer rank among the most important industrial fermentation process parameters. The present study analyzes the power input and gas hold-up as a function of the flow regime, impeller diameter, and rheological properties in a pilot scale reactor (160 L) equipped with four Rushton impellers. This leads to four dimensionless numbers for predicting measurements in pilot and industrial bioreactors (110 and 170 m3) with a standard deviation of 7 % to 29 %. This is unparalleled for the underlying aerated and non-Newtonian fermentation broths. Several existing correlation equations are discussed to be dissatisfying (up to 130 % deviation), and might be sufficiently valid only within scale or for small scaling factors. The introduced approach predicts adequately accurate over three orders of magnitude. Based on these encouraging results, we identified the Galilei number and the power concept as the central elements in combination with the consequent dimensional analysis for an efficient scaling betweeen pilot and industrial scale.

Multi-Statistical Approach for the Study of Volatile Compounds of Industrial Spoiled Manzanilla Spanish-Style Table Olive Fermentations

Table olives can suffer different types of spoilage during fermentation. In this work, a multi-statistical approach (standard and compositional data analysis) was used for the study of the volatile organic compounds (VOCs) associated with altered (butyric, sulfidic, and putrid) and non-altered (normal) Manzanilla Spanish-style table olive fermentations. Samples were collected from two industrial fermentation yards in Seville (Spain) in the 2019/2020 season. The VOC profiles of altered (n = 4) and non-altered (n = 6) samples were obtained by headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Ninety-one VOCs were identified and grouped into alcohols (30), esters (21), carbonyl compounds (12), acids (10), terpenes (6), phenols (6), sulfur compounds (2), and others (4). The association of the VOCs with spoilage samples depended on the standard or compositional statistical methodology used. However, butyric spoilage was strongly linked by several techniques to methyl butanoate, ethyl butanoate, and butanoic acid; sulfidic spoilage with 2-propyl-1-pentanol, dimethyl sulfide, methanol, 2-methylbutanal, 2-methyl-2-butenal, ethanol, 2-methyl-3-buten-2-ol, and isopentanol, while putrid was mainly related to D-limonene and 2-pentanol. Our data contribute to a better characterisation of non-zapatera spoiled table olive fermentations and show the convenience of using diverse statistical techniques for a most robust selection of spoilage VOC markers.

Yeast Morphology Assessment through Automated Image Analysis during Fermentation

The kinetics and success of an industrial fermentation are dependent upon the health of the microorganism(s) responsible. Saccharomyces sp. are the most commonly used organisms in food and beverage production; consequently, many metrics of yeast health and stress have been previously correlated with morphological changes to fermentations kinetics. Many researchers and industries use machine vision to count yeast and assess health through dyes and image analysis. This study assessed known physical differences through automated image analysis taken throughout ongoing high stress fermentations at various temperatures (30 °C and 35 °C). Measured parameters included sugar consumption rate, number of yeast cells in suspension, yeast cross-sectional area, and vacuole cross-sectional area. The cell morphological properties were analyzed automatically using ImageJ software and validated using manual assessment. It was found that there were significant changes in cell area and ratio of vacuole to cell area over the fermentation. These changes were temperature dependent. The changes in morphology have implications for rates of cellular reactions and efficiency within industrial fermentation processes. The use of automated image analysis to quantify these parameters is possible using currently available systems and will provide additional tools to enhance our understanding of the fermentation process.