Removal of phenolic compounds from sugarcane syrup and impact on Saccharomyces cerevisiae fermentation for ß-farnesene production.

This work aimed to study for the first time the effects of phenolic compounds from sugarcane syrup on Saccharomyces cerevisiae ß-farnesene fermentation by removing them from this feedstock. Syrup purification was optimized through a central composite design using five types of activated charcoal: three contact times (1-24 h) and three adsorbent concentrations (10-150 g L-1 ). The optimal purification condition-charcoal pellets at 115 g L-1 and contact time of 12.5 h-led to 96.7% of phenolic compounds removal and 43.7% of syrup recovery. The effects of reducing phenolic content from approximately 7.0-0.3 mg L-1 in sugarcane syrup on yeast fermentation varied with the scale. An increase in biomolecule productivity was only observed in shake-flasks (11%) and in biomass productivity only in the 2 L bioreactor (12%). Thus, phenolic compounds from sugarcane syrup do not influence ß-farnesene production at a large scale under the conditions tested.

Exploring yeast-based microbial interactions: The next frontier in postharvest biocontrol.

Fresh fruits and vegetables are susceptible to a large variety of spoilage agents before and after harvest. Among these, fungi are mostly responsible for the microbiological deteriorations that lead to economically significant losses of fresh produce. Today, synthetic fungicides represent the first approach for controlling postharvest spoilage in fruits and vegetables worldwide. However, the emergence of fungicide-resistant pathogen biotypes and the increasing awareness of consumers toward the health implications of hazardous chemicals imposed an urgent need to reduce the use of synthetic fungicides in the food supply; this phenomenon strengthened the search for alternative biocontrol strategies that are more effective, safer, nontoxic, low-residue, environment friendly, and cost-effective. In the last decade, biocontrol with antagonistic yeasts became a promising strategy to reduce chemical compounds during fruit and vegetable postharvest, and several yeast-based biocontrol products have been commercialized. Biocontrol is a multipartite system that includes different microbial groups (spoilage mold, yeast, bacteria, and nonspoilage resident microorganisms), host fruit, vegetables, or plants, and the environment. The majority of biocontrol studies focused on yeast-mold mechanisms, with little consideration for yeast-bacteria and yeast-yeast interactions. The current review focused mainly on the unexplored yeast-based interactions and the mechanisms of actions in biocontrol systems as well as on the importance and advantages of using yeasts as biocontrol agents, improving antagonist efficiency, the commercialization process and associated challenges, and future perspectives.

Peptide extract from spent yeast improves resistance of Saccharomyces cerevisiae to oxidative stress.

Yeast cells face various stress factors during industrial fermentations, since they are exposed to harsh environmental conditions, which may impair biomolecules productivity and yield. In this work, the use of an antioxidant peptide extract obtained from industrial spent yeast was explored as supplement for Saccharomyces cerevisiae fermentation to prevent a common bottleneck: oxidative stress. For that, a recombinant yeast strain, producer of ß-farnesene, was firstly incubated with 0.5 and 0.7 g/L peptide extract, in the presence and absence of hydrogen peroxide (an oxidative stress inducer), for 1-5 h, and then assayed for intracellular reactive oxygen species, and growth ability in agar spot assays. Results showed that under 2 mM H2O2, the peptide extract could improve cells growth and reduce reactive oxygen species production. Therefore, this antioxidant effect was further evaluated in shake-flasks and 2-L bioreactor batch fermentations. Peptide extract (0.7 g/L) was able to increase yeast resistance to the oxidative stress promoted by 2 mM H2O2, by reducing reactive oxygen species levels between 1.2- and 1.7-fold in bioreactor and between 1.2- and 3-fold in shake-flask fermentations. Moreover, improvements on yeast cell density of up to 1.5-fold and 2-fold, and on biomolecule concentration of up to 1.6-fold and 2.8-fold, in bioreactor and shake-flasks, respectively, were obtained. Thus, culture medium supplementation with antioxidant peptide extracted from industrial spent yeast is a promising strategy to improve fermentation performance while valuing biomass waste. This valorization can promote a sustainable and eco-friendly solution for the biotechnology industry by the implementation of a circular economy model. KEY POINTS: • Peptide extract from spent yeast applied for the first time on yeast fermentation. • Antioxidant peptide extract enhanced S. cerevisiae oxidative stress resistance. • Fermentation performance under stress improved by peptide extract supplementation.

Fermentation Strategies for Production of Pharmaceutical Terpenoids in Engineered Yeast.

Terpenoids, also known as isoprenoids, are a broad and diverse class of plant natural products with significant industrial and pharmaceutical importance. Many of these natural products have antitumor, anti-inflammatory, antibacterial, antiviral, and antimalarial effects, support transdermal absorption, prevent and treat cardiovascular diseases, and have hypoglycemic activities. Production of these compounds are generally carried out through extraction from their natural sources or chemical synthesis. However, these processes are generally unsustainable, produce low yield, and result in wasting of substantial resources, most of them limited. Microbial production of terpenoids provides a sustainable and environment-friendly alternative. In recent years, the yeast Saccharomyces cerevisiae has become a suitable cell factory for industrial terpenoid biosynthesis due to developments in omics studies (genomics, transcriptomics, metabolomics, proteomics), and mathematical modeling. Besides that, fermentation development has a significant importance on achieving high titer, yield, and productivity (TYP) of these compounds. Up to now, there have been many studies and reviews reporting metabolic strategies for terpene biosynthesis. However, fermentation strategies have not been yet comprehensively discussed in the literature. This review summarizes recent studies of recombinant production of pharmaceutically important terpenoids by engineered yeast, S. cerevisiae, with special focus on fermentation strategies to increase TYP in order to meet industrial demands to feed the pharmaceutical market. Factors affecting recombinant terpenoids production are reviewed (strain design and fermentation parameters) and types of fermentation process (batch, fed-batch, and continuous) are discussed.

Lipids by Yarrowia lipolytica Strains Cultivated on Glucose in Batch Cultures.

Oleaginous microorganisms, such as Yarrowia lipolytica, accumulate lipids that can have interesting applications in food biotechnology or the synthesis of biodiesel. Y. lipolytica yeast can have many advantages such as wide substrate range usage and robustness to extreme conditions, while under several culture conditions it can produce high lipid productivity. Based on this assumption, in this study, 12 different Yarrowia lipolytica strains were used to investigate microbial lipid production using a glucose-based medium under nitrogen-limited conditions in shake-flask cultivations. Twelve wild-type or mutant strains of Yarrowia lipolytica which were newly isolated or belonged to official culture collections were tested, and moderate lipid quantities (up to 1.30 g/L) were produced; in many instances, nitrogen limitation led to citric acid production in the medium. Lipids were mainly composed of C16 and C18 fatty acids. Most of the fatty acids of the microbial lipid were unsaturated and corresponded mainly to oleic, palmitic and linoleic acids. Linolenic acid (C18:3) was produced in significant quantities (between 10% and 20%, wt/wt of dry cell weight (DCW)) by strains H917 and Po1dL.

Screening various Yarrowia lipolytica strains for citric acid production.

Citric acid (CA) productivity by Yarrowia lipolytica dependents on strain type, carbon source, carbon to nitrogen (C/N) molar ratio as well as physicochemical conditions (pH, temperature, oxygen transfer rate, etc.). In the current study, 10 different Y. lipolytica strains were first challenged in shake-flask culture for CA production in a glucose-based media under nitrogen-limited conditions. For the most promising one, strain K57, CA productivity was monitored during culture in batch bioreactor for three initial C/N molar ratio (167, 367, and 567 Cmol/Nmol). The highest CA yield (0.77 g/g glucose), titre (72.3 g/L CA), and productivity (0.04 g/g.h) were found for C/N ratio of 367. However, the highest growth rate was obtained for an initial C/N ratio of 167. From these results, Y. lipolytica strain K57 could be considered to produce CA at higher titre on glucose-based medium in batch bioreactor than others Y. lipolytica strain reported in the literature.