Transcriptomics to evaluate the influence mechanisms of ethanol on the ester production of Wickerhamomyces anomalus with the induction of lactic acid.

Wickerhamomyces anomalus is one of the most important ester-producing strains in Chinese baijiu brewing. Ethanol and lactic acid are the main metabolites produced during baijiu brewing, but their synergistic influence on the growth and ester production of W. anomalus is unclear. Therefore, in this paper, based on the contents of ethanol and lactic acid during Te-flavor baijiu brewing, the effects of different ethanol concentrations (3, 6, and 9% (v/v)) combined with 1% lactic acid on the growth and ester production of W. anomalus NCUF307.1 were studied and their influence mechanisms were analyzed by transcriptomics. The results showed that the growth of W. anomalus NCUF307.1 under the induction of lactic acid was inhibited by ethanol. Although self-repair mechanism of W. anomalus NCUF307.1 induced by lactic acid was initiated at all concentrations of ethanol, resulting in significant up-regulation of genes related to the Genetic Information Processing pathway, such as cell cycle-yeast, meiosis-yeast, DNA replication and other pathways. However, the accumulation of reactive oxygen species and the inhibition of pathways associated with carbohydrate and amino acid metabolism may be the main reason for the inhibition of growth in W. anomalus NCUF307.1. In addition, 3% and 6% ethanol combined with 1% lactic acid could promote the ester production of W. anomalus NCUF307.1, which may be related to the up-regulation of EAT1, ADH5 and TGL5 genes, while the inhibition in 9% ethanol may be related to down-regulation of ATF2, EAT1, ADH2, ADH5, and TGL3 genes.

Contribution of trehalose to ethanol stress tolerance of Wickerhamomyces anomalus.

BACKGROUND: The ascomycetous heterothallic yeast Wickerhamomyces anomalus (WA) has received considerable attention and has been widely reported in the winemaking industry for its distinctive physiological traits and metabolic attributes. An increased concentration of ethanol during ethanol fermentation, however, causes ethanol stress (ES) on the yeast cells. Trehalose has been implicated in improving survival under various stress conditions in microorganisms. Herein, we determined the effects of trehalose supplementation on the survival, differentially expressed genes (DEGs), cellular morphology, and oxidative stress tolerance of WA in response to ES. RESULTS: The results indicated that trehalose improved the survival and anomalous surface and ultrastructural morphology of WA. Additionally, trehalose improved redox homeostasis by reducing the levels of reactive oxygen species (ROS) and inducing the activities of antioxidant enzymes. In addition, DEGs affected by the application of trehalose were enriched in these categories including in gene expression, protein synthesis, energy metabolism, and cell cycle pathways. Additionally, trehalose increased the content of intracellular malondialdehyde (MDA) and adenosine triphosphate. CONCLUSIONS: These results reveal the protective role of trehalose in ES mitigation and strengthen the possible uses of WA in the wine fermentation sector.

Kinetics and mathematical model of killer/sensitive interaction under different physicochemical conditions of must/wine: a study from a biological point of view.

Fermentation of grape must to wine is carried out by a complex microbial mixture, which also involves spoilage yeasts of wine. The latter yeasts produce organoleptic changes that cause significant economic losses to the wine industry. SO2 is traditionally used to control this spoilage populations, but because of its harmful effects on human health, biocontrol has emerged as an alternative treatment. Although studies have been carried out to select biocontroller yeasts and examine their underlying mechanisms of action, reports on their application have not been published yet. To better understand the interaction and the successful application of biocontrol, the use of mathematical models, among other methods, is important, as they facilitate the prediction of success or failure of the antagonist. The objective of the present study was to use an existing mathematical model to obtain information about the yeast's interaction assayed and to validate its predictive use under different physicochemical conditions during the wine fermentation, and eventually predict biocontrol kinetics. The mathematical model was applied to the fermentation conditions and provided information on the kinetic parameters of the biocontrol interaction and allowed interpretations about other parameters. The model was applied in the different physicochemical conditions for the biocontrol and did not fit correctly to experimental data, and therefore an improvement was proposed which was successful and presented new hypotheses.

Efficacy and mechanism of action of food isolated yeasts in the control of Aspergillus flavus growth on pistachio nuts.

Due to its nutritional characteristics, dried fruit and in particular pistachio is considered an important component in the daily diet. Unfortunately, pistachio nuts can be contaminated during storage with a wide range of pathogenic fungi, including Aspergillus flavus. The present work has evaluated how the use of two yeast strains belonging to the species Wickerhamomyces anomalus and Metschnikowia pulcherrima can inhibit the in vitro and in vivo growth of A. flavus. Both yeast strains demonstrated a good in vitro antifungal activity attributable to a specific mechanism of action, although higher efficacy was evidenced by W. anomalus strain. The production of volatile organic compounds (VOCs) and lytic enzymes was hypothesized as the main mechanisms of action exerted by W. anomalus, while the nutritional competition for iron was assumed as the main biocontrol mechanism for M. pulcherrima. Moreover, from the results of the in vivo test carried out on artificially infected pistachio seeds, it was clear as M. pulcherrima strain showed the same efficacy of W. anomalus in inhibiting the growth and sporulation of A. flavus mold, despite W. anomalus was the most effective strain during in vitro assay. Altogether, these results indicate that both strains could be considered as potential biocontrol agents against A. flavus fungal growth, notwithstanding it is always important considering the tritrophic interaction (yeast-mold- host), which could play a crucial role in determining the final results.

Effects of Yeast Species and Processing on Intestinal Health and Transcriptomic Profiles of Atlantic Salmon (Salmo salar) Fed Soybean Meal-Based Diets in Seawater.

The objective of the current study was to examine the effects of yeasts on intestinal health and transcriptomic profiles from the distal intestine and spleen tissue of Atlantic salmon fed SBM-based diets in seawater. Cyberlindnera jadinii (CJ) and Wickerhamomyces anomalus (WA) yeasts were heat-inactivated with spray-drying (ICJ and IWA) or autolyzed at 50 °C for 16 h (ACJ and AWA), followed by spray-drying. Six diets were formulated, one based on fishmeal (FM), a challenging diet with 30% soybean meal (SBM) and four other diets containing 30% SBM and 10% of each of the four yeast fractions (i.e., ICJ, ACJ, IWA and AWA). The inclusion of CJ yeasts reduced the loss of enterocyte supranuclear vacuolization and reduced the population of CD8α labeled cells present in the lamina propria of fish fed the SBM diet. The CJ yeasts controlled the inflammatory responses of fish fed SBM through up-regulation of pathways related to wound healing and taurine metabolism. The WA yeasts dampened the inflammatory profile of fish fed SBM through down-regulation of pathways related to toll-like receptor signaling, C-lectin receptor, cytokine receptor and signal transduction. This study suggests that the yeast species, Cyberlindnera jadinii and Wickerhamomyces anomalus are novel high-quality protein sources with health-beneficial effects in terms of reducing inflammation associated with feeding plant-based diets to Atlantic salmon.

Use of Pineapple Waste as Fuel in Microbial Fuel Cell for the Generation of Bioelectricity.

The excessive use of fossil sources for the generation of electrical energy and the increase in different organic wastes have caused great damage to the environment; these problems have promoted new ways of generating electricity in an eco-friendly manner using organic waste. In this sense, this research uses single-chamber microbial fuel cells with zinc and copper as electrodes and pineapple waste as fuel (substrate). Current and voltage peaks of 4.95667 ± 0.54775 mA and 0.99 ± 0.03 V were generated on days 16 and 20, respectively, with the substrate operating at an acid pH of 5.21 ± 0.18 and an electrical conductivity of 145.16 ± 9.86 mS/cm at two degrees Brix. Thus, it was also found that the internal resistance of the cells was 865.845 ± 4.726 Ω, and a maximum power density of 513.99 ± 6.54 mW/m2 was generated at a current density of 6.123 A/m2, and the final FTIR spectrum showed a clear decrease in the initial transmittance peaks. Finally, from the biofilm formed on the anodic electrode, it was possible to molecularly identify the yeast Wickerhamomyces anomalus with 99.82% accuracy. In this way, this research provides a method that companies exporting and importing this fruit may use to generate electrical energy from its waste.

A Comparative Study of Various Pretreatment Approaches for Bio-Ethanol Production from Willow Sawdust, Using Co-Cultures and Mono-Cultures of Different Yeast Strains.

The effect of different pretreatment approaches based on alkali (NaOH)/hydrogen peroxide (H2O2) on willow sawdust (WS) biomass, in terms of delignification efficiency, structural changes of lignocellulose and subsequent fermentation toward ethanol, was investigated. Bioethanol production was carried out using the conventional yeast Saccharomyces cerevisiae, as well as three non-conventional yeasts strains, i.e., Pichia stipitis, Pachysolen tannophilus, Wickerhamomyces anomalus X19, separately and in co-cultures. The experimental results showed that a two-stage pretreatment approach (NaOH (0.5% w/v) for 24 h and H2O2 (0.5% v/v) for 24 h) led to higher delignification (38.3 ± 0.1%) and saccharification efficiency (31.7 ± 0.3%) and higher ethanol concentration and yield. Monocultures of S. cerevisiae or W. anomalus X19 and co-cultures with P. stipitis exhibited ethanol yields in the range of 11.67 ± 0.21 to 13.81 ± 0.20 g/100 g total solids (TS). When WS was subjected to H2O2 (0.5% v/v) alone for 24 h, the lowest ethanol yields were observed for all yeast strains, due to the minor impact of this treatment on the main chemical and structural WS characteristics. In order to decide which is the best pretreatment approach, a detailed techno-economical assessment is needed, which will take into account the ethanol yields and the minimum processing cost.

Assessment of chemical constitution and aroma properties of kiwi wines obtained from pure and mixed fermentation with Wickerhamomyces anomalus and Saccharomyces cerevisiae.

BACKGROUND: To improve the aroma of kiwi wine through the utilization of Wickerhamomyces anomalus, kiwi juice was fermented using a selected W. anomalus strain in pure culture and mixed fermentations with Saccharomyces cerevisiae, which was inoculated simultaneously and sequentially. The physicochemical indices, volatile compounds and aroma properties of the kiwi wines were assessed. RESULTS: The study suggested that the ethanol, color indices and organic acids of the wines were closely related to the method of inoculation. Compared with the pure S. cerevisiae fermentation, the mixed fermentations produced more varieties and concentrations of volatiles. The sequential fermentations increased the concentrations of esters and terpenes, improving the flower and sweet fruit notes of the wines. The simultaneous inoculation enhanced the contents of esters and aldehydes, intensifying the flower, sweet and sour fruit of the wines. Partial least-squares regression analysis showed that esters and terpenes contributed greatly to the flower and sweet fruit aroma, whereas aldehydes were the major contributors to the sour note. CONCLUSION: Based on our results, the mixed fermentations not only enriched the types and concentrations of volatiles, but also had better sensory properties. © 2021 Society of Chemical Industry.

Antimicrobial and prebiotic activity of mannoproteins isolated from conventional and nonconventional yeast species-the study on selected microorganisms.

Yeast mannoproteins are proposed as a paraprobiotics with antimicrobial and prebiotic properties. They can be used as biopreservatives in food and in diseases therapies. The knowledge about the specificity and/or capability of their influence on the growth of different microorganism is limited. The study determined the effect of mannoprotein preparations of Saccharomyces cerevisiae (S. cerevisiae) ATCC 7090 and nonconventional yeast origin [Metschnikowia reukaufii (M. reukaufii) WLP 4650 and Wickerhamomyces anomalus (W. anomalus) CCY 38-1-13] on the growth of selected bacteria of the genera: Lactobacilllus, Limosilatobacillus, Limosilatobacillus, Bifidobacterium, Staphylococcus, Enterococcus, Pseudomonas, Escherichia, Proteus and Salmonella. The degree of stimulation or growth inhibition of tested bacteria depended on the type and dose of the mannoprotein and the bacterial strain. The addition of the tested preparations in the entire range of applied concentrations had a positive effect especially on the growth of Lactobacillus arabinosus ATCC 8014 and Bifidobacterium animalis subsp. lactis B12. Mannoproteins isolated from S. cerevisiae limited the growth of the Escherichia coli (E. coli) ATCC 25922, Pseudomonas aureoginosa (P. aureoginosa) ATCC 27853, Proteus mirabilis ATCC 35659 and Salmonella Enteritidis ATCC 13076 to the greatest extent, while preparations of M. reukaufii and W. anomalus origin most effectively limited the growth of Staphylococcus aureus strains, E. coli and P. aureoginosa. The growth of Enterococcus faecalis was stimulated by the presence of all studied preparations in most of the concentrations used. Further research will determine how the purification process of studied mannoproteins or oligosaccharide fractions, its structure and composition influence on the growth of selected bacteria and what is the mechanism of its activity.

Microbial Application to Improve Olive Mill Wastewater Phenolic Extracts.

Olive mill wastewater (OMW) contains valuable and interesting bioactive compounds, among which is hydroxytyrosol, which is characterized by a remarkable antioxidant activity. Due to the health claims related to olive polyphenols, the aim of this study was to obtain an extract from OMW with an increased level of hydroxytyrosol by means of microbial enzymatic activity. For this purpose, four commercial adsorbent resins were selected and tested. The beta-glucosidase and esterase activity of strains of Wickerhamomyces anomalus, Lactiplantibacillus plantarum, and Saccharomyces cerevisiae were also investigated and compared to those of a commercial enzyme and an Aspergillus niger strain. The W. anomalus strain showed the best enzymatic performances. The SP207 resin showed the best efficiency in selective recovery of hydroxytyrosol, tyrosol, oleuropein, and total phenols. The bioconversion test of the OMW extract was assessed by using both culture broths and pellets of the tested strains. The results demonstrated that the pellets of W. anomalus and L. plantarum were the most effective in hydroxytyrosol increasing in phenolic extract. The interesting results suggest the possibility to study new formulations of OMW phenolic extracts with multifunctional microorganisms.

The killer yeast Wickerhamomyces anomalus Cf20 exerts a broad anti-Candida activity through the production of killer toxins and volatile compounds.

Candidiasis is a group of opportunistic infections caused by yeast of the genus Candida. The appearance of drug resistance and the adverse effects of current antifungal therapies require the search for new, more efficient therapeutic alternatives. Killer yeasts have aroused as suitable candidates for mining new antifungal compounds. Killer strains secrete antimicrobial proteins named killer toxins, with promissory antifungal activity. Here we found that the killer yeast Wickerhamomyces anomalus Cf20 and its cell-free supernatant (CFS) inhibited six pathogenic strains and one collection strain of Candida spp. The inhibition is mainly mediated by secreted killer toxins and, to a lesser extent, by volatile compounds such as acetic acid and ethyl acetate. A new large killer toxin (>180 kDa) was purified, which exerted 70-74% of the total CFS anti-Candida activity, and the previously described glucanase KTCf20 was inhibitory in a lesser extent as well. In addition, we demonstrated that Cf20 possesses the genes encoding for the ß-1,3-glucanases WaExg1 and WaExg2, proteins with extensively studied antifungal activity, particularly WaExg2. Finally, the 10-fold concentrated CFS exerted a high candidacidal effect at 37°C, completely inhibiting the fungal growth, although the nonconcentrated CFS (RCF 1) had very limited fungistatic activity at this temperature. In conclusion, W. anomalus Cf20 produces different low and high molecular weight compounds with anti-Candida activity that could be used to design new therapies for candidiasis and as a source for novel antimicrobial compounds as well.

Antibiotic Activity of Wickerhamomyces anomalus Mycocins on Multidrug-Resistant Acinetobacter baumannii.

To evaluate the susceptibility of multidrug-resistant Acinetobacter baumannii to mycocins produced by Wickerhamomyces anomalus and to verify the cytotoxicity of these compounds. Three culture supernatants of W. anomalus (WA40, WA45, and WA92), containing mycocins (WA40M1, WA45M2, and WA92M3), were tested on A. baumannii using broth microdilution methods, solid medium tests, and cytotoxicity tests in human erythrocytes and in Artemia saline Leach. W. anomalus was able to produce high antimicrobial mycocins, as even at high dilutions, they inhibited A. baumannii. In a solid medium, it was possible to observe the inhibition of A. baumannii, caused by the diffusion of mycocins between agar. Finally, the three supernatants were not cytotoxic when tested on human erythrocytes and Artemia salina. According to the evidence in this study, the mycocins of W. anomalus have been effective and could be used in the development of new antimicrobial substances.

Microbial transformation of diosgenin to diosgenone by Wickerhamomyces anomalus JQ-1 obtained from Naxi traditional Jiu Qu.

Diosgenone [(20S,22R,25R)-spirost-4-en-3-one, C27H40O3] has been considered as a potential therapeutic alternative remedy for malaria. An efficient and economical approach of microbial transformation with diosgenin to diosgenone by the yeast strain Wickerhamomyces anomalus JQ-1 from Naxi traditional Jiu Qu was developed in this study. Chromatographic analysis confirmed that 85% of 0.1 mM diosgenin was transformed to diosgenone within 72 h. This research demonstrates that diosgenin could be converted to diosgenone through two-step pathway including 3ß-hydroxyl oxidation and double bond isomerization rather than through one-step pathway, which prompted a further inference that the oxidation activity in W. anomalus JQ-1 has the same function with the Oppenauer-type oxidation which can convert diosgenin into diosgenone. Gaining specific functional strains from traditional fermented products will be a potential direction and ethnobotanical researches could provide helps with discovery and utilization of microbial resources.

Contribution of autochthonous yeasts with probiotic potential to the aroma profile of fermented Guajillo pepper sauce.

BACKGROUND: Three yeast strains with probiotic potential, Hanseniaspora opuntiae, Pichia kudriavzevii, and Wickerhamomyces anomalus were inoculated in the fermentation of Guajillo chilli pepper (Capsicum annuum L.) sauce, and the different aroma profiles were investigated. Using headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) analysis and gas chromatography-olfactometry (GCO), flavour compound production was evaluated during the fermentation of the Guajillo chilli pepper sauces. RESULTS: A total of 78 volatile compounds were identified during the yeast fermentation of the sauce. Most aldehydes and terpenes detected were present at the beginning of the fermentation, indicating a Guajillo chilli pepper origin. Among the 34 active aroma compounds detected by GCO, propanoic acid (cheesy), 3-methylbutanoic acid (sharp, cheese), ethyl 2-methylbutanoate (fruity), and 6-methyl-5-hepten-2-one (strong, citrus) were identified as key aroma contributors produced by the inoculation of the yeasts. A different aroma profile was produced by probiotic yeast. Hanseniaspora opuntiae produced an aroma profile with herbal and green notes based on high production of aldehydes, ketones, and acetic acid. Pichia kudriavzevii and W. anomalus produced fruity, green-herbal, and cheesy notes based on ester compounds, alcohol and branched-chain acids production although, the production of propanoic acid by W. anomalus increased the cheesy character in the sauces. CONCLUSION: The aroma profile of fermented chilli pepper sauces depends not only on the chili pepper varieties used but also on the fermentation process as a source of aroma compounds. The use of probiotic yeast can be used to improve and diversify the aroma profile of fermented chilli pepper sauces. © 2020 Society of Chemical Industry.

High-affinity transport, cyanide-resistant respiration, and ethanol production under aerobiosis underlying efficient high glycerol consumption by Wickerhamomyces anomalus.

Wickerhamomyces anomalus strain LBCM1105 was originally isolated from the wort of cachaça (the Brazilian fermented sugarcane juice-derived Brazilian spirit) and has been shown to grow exceptionally well at high amounts of glycerol. This paramount residue from the biodiesel industry is a promising cheap carbon source for yeast biotechnology. The assessment of the physiological traits underlying the W. anomalus glycerol consumption ability in opposition to Saccharomyces cerevisiae is presented. A new WaStl1 concentrative glycerol-H+ symporter with twice the affinity of S. cerevisiae was identified. As in this yeast, WaSTL1 is repressed by glucose and derepressed/induced by glycerol but much more highly expressed. Moreover, LBCM1105 aerobically growing on glycerol was found to produce ethanol, providing a redox escape to compensate the redox imbalance at the level of cyanide-resistant respiration (CRR) and glycerol 3P shuttle. This work is critical for understanding the utilization of glycerol by non-Saccharomyces yeasts being indispensable to consider their industrial application feeding on biodiesel residue.

Edible coatings incorporating pomegranate peel extract and biocontrol yeast to reduce Penicillium digitatum postharvest decay of oranges.

This study investigated the potential use of two edible coatings, chitosan (CH) and locust bean gum (LBG), which incorporated chemically characterized water pomegranate peel extract (WPPE) or methanol pomegranate peel extract (MPPE) and the biocontrol agent (BCA) Wickerhamomyces anomalus, to control the growth of Penicillium digitatum and to reduce the postharvest decay of oranges. CH and LBG including pomegranate peel extracts (PPEs) at different concentrations were tested in vitro against P. digitatum to determine their antifungal efficacy; at the same time, the tolerance of viable cells of W. anomalus to increasing concentrations of WPPE and MPPE extracts was assessed. The potential application of selected bioactive coatings was evaluated in vivo on oranges, which had been artificially inoculated with P. digitatum, causal agent of green mold decay. CH incorporating MPPE or WPPE at all concentrations was able to inhibit in vitro P. digitatum, while LBG was active only at the highest MPPE or WPPE concentrations. W. anomalus BS91 was slightly inhibited only by MPPE-modified coatings, while no inhibition was observed by WPPE, which was therefore selected for the in vivo trials on oranges artificially inoculated with P. digitatum. The experimental results proved that the addition of 0.361 g dry WPPE/mL, both to CH and LBG coatings, significantly reduced disease incidence (DI) by 49 and 28% respectively, with respect to the relative controls. Besides the combination CH or LBG + WPPE, the addition of W. anomalus cells to coatings strengthened the antifungal effect with respect to the relative controls, as demonstrated by the significant reduction of DI (up to 95 and 75% respectively). The findings of the study contribute to the valorization of a value-added industrial byproduct and provide a significant advancement in the development of new food protectant formulations, which benefit from the synergistic effect between biocontrol agents and natural bioactive compounds.

Antifungal activity and expression patterns of extracellular chitinase and ß-1,3-glucanase in Wickerhamomyces anomalus EG2 treated with chitin and glucan.

In this study, the expression patterns of extracellular chitinase and ß-1,3-glucanase from cultured Wickerhamomyces anomalus EG2 treated with chitin, glucan, and chemical chitinase inhibitors (kinetin, caffeine, and acetazolamide) were investigated using SDS-PAGE. Relationship between enzyme expression and antifungal activity from yeast plays a very important role for biocontrol of phytopathoges. To determine antifungal activity against phytopathogens, W. anomalus EG2 was shown to strongly inhibit hyphal growth of Fusarium oxysporum KACC 40032 and Rhizoctonia solani KACC 40111. Slight chitinase activity was observed 12 h after incubation in both PDB and YPD medium without colloidal chitin. The molecular weight of chitinase was approximately 124 kDa ß-1,3-Glucanase isoenzyme (GN1 and GN2) was observed distinctly on SDS-PAGE gels when laminarin was used as a substrate. ß-1,3-Glucanase isoenzyme was not observed when using glucan-containing high polymer complex (GHPC) as a substrate. Production of chitinase from W. anomalus EG2 was inhibited slightly by acetazolamide. Abnormal and cluster-shaped cells of W. anomalus EG2 were observed in both PDB and YPD medium treated with colloidal chitin. These results indicated that W. anomalus EG2 could be applied commercially as a biological control agent of phytopathogens and as a bioinhibitor of yeast cell growth.

Urea production by yeasts other than Saccharomyces in food fermentation.

Urea is an important intermediate in the synthesis of carcinogenic ethyl carbamate in various food fermentations. Identifying urea-producing microorganisms can help control or reduce ethyl carbamate production. Using Chinese liquor fermentation as a model system, we identified the yeasts responsible for urea production. Urea production was positively correlated to the yeast population (R = 0.523, P = 0.045), and using high-throughput sequencing, we identified 26 yeast species. Partial least squares regression and correlation analysis indicated that Wickerhamomyces anomalus was the most important yeast to produce urea (variable importance plot = 1.927; R = 0.719, P = 0.002). Besides, we found that in W. anomalus the CAR1 gene (responsible for urea production) was 67% identical to that of Saccharomyces cerevisiae. Wickerhamomyces anomalus produced more urea (910.0 µg L-1) than S. cerevisiae (300.1 µg L-1). Moreover, urea production increased to 1261.2 µg L-1 when the two yeasts were co-cultured in a simulated fermentation, where the transcription activity of the CAR1 gene increased by 140% in W. anomalus and decreased by 40% in S. cerevisiae. Our findings confirm that a yeast other than Saccharomyces, namely W. anomalus, contributes more to urea formation in a simulated sorghum fermentation. These findings provide the basis for strategies to control or reduce ethyl carbamate formation.

Postharvest biocontrol ability of killer yeasts against Monilinia fructigena and Monilinia fructicola on stone fruit.

The antagonistic effects of Debaryomyces hansenii KI2a, D. hansenii MI1a and Wickerhamomyces anomalus BS91 were tested against Monilinia fructigena and Monilinia fructicola in in vitro and in vivo trials. All yeast strains demonstrated antifungal activity at different levels depending on species, strain and pathogen. D hansenii KI2a and W. anomalus BS91 showed the highest biocontrol activity in vitro; the production of hydrolytic enzymes, killer toxins and volatile organic compounds (VOCs) were hypothesized as their main mechanisms of action against pathogens. D hansenii KI2a and W. anomalus BS91 significantly reduced brown rot incidence and severity on peach and plum fruits artificially inoculated with M. fructigena and M. fructicola, especially when applied 24 h before pathogen inoculation. On the opposite, D. hansenii MI1a exhibited weak antagonistic activity towards M. fructigena on peach and plum fruits and was ineffective against M. fructicola. The noticeable ability of W. anomalus BS91 to control brown rot could be also correlated with its high capacity to colonize the wound tissue and to increase its population density. Accordingly, the antagonistic strains of D. hansenii and W. anomalus could be proposed as active ingredients for the development of biofungicides against Monilinia species that are responsible for considerable economic losses in stone fruit crops.

Lipid production by yeasts grown on crude glycerol from biodiesel industry.

The main carbon source used for growth by four yeast strains (Yarrowia lipolytica CCMA 0357, Y. lipolytica CCMA 0242, Wickerhamomyces anomalus CCMA 0358, and Cryptococcus humicola CCMA 0346) and their lipid production were evaluated, using different concentrations of crude and pure glycerol and glucose. Whereas crude glycerol (100 g/L) was the main carbon source used by Y. lipolytica CCMA 0357 (nearly 15 g/L consumed at 120 hr) and W. anomalus CCMA 0358 (nearly 45.10 g/L consumed at 48 hr), pure glycerol (150 g/L) was the main one used by C. humicola CCMA 0346 (nearly 130 g/L consumed). On the other hand, Y. lipolytica CCMA 0242 used glucose (100 g/L) as its main source of carbon (nearly 96.48 g/L consumed). Y. lipolytica CCMA 0357 demonstrated the highest lipid production [about 70% (wt/wt)], forming palmitic (45.73% of fatty acid composition), stearic (16.43%), palmitoleic (13.29%), linolenic (10.77%), heptadecanoic (4.07%), and linoleic (14.14%) acids. Linoleic acid, an essential fatty acid, was produced by all four yeast strains but in varying degrees, representing 70.42% of the fatty acid profile of lipids produced by C. humicola CCMA 0346.