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Antagonistic yeasts suppress plant pathogenic fungi by various mechanisms, but their biocontrol efficacy also depends on the ability to compete and persist in the environment. The goal of the work presented here was to quantify the composition of synthetic yeast communities in order to determine the competitiveness of different species and identify promising candidates for plant protection. For this purpose, colony counting of distinct species and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS; MALDI biotyping) were used to distinguish different yeast species and to quantify the composition of a synthetic community of six yeasts (Aureobasidium pullulans, Candida subhashii, Cyberlindnera sargentensis, Hanseniaspora sp., Metschnikowia pulcherrima and Pichia kluyveri) over time, on apples and in soil, and in different growth media. These studies revealed important characteristics that predispose the different species for particular applications. For example, the competitiveness and antagonistic activity of C. subhashii was strongly increased in the presence of N‐acetylglucosamin as the sole carbon source, M. pulcherrima and A. pullulans were the strongest competitors on apple, and C. sargentensis competed the best in soil microcosms. Based on these laboratory studies, M. pulcherrima and A. pullulans are promising candidates for biocontrol applications against fungal phyllosphere diseases, while C. sargentensis may hold potential for use against soilborne fungal pathogens. These results document the potential of MALDI‐TOF MS for the quantitative analysis of synthetic yeast communities and highlight the value of studying microorganisms with relevant functions in moderately complex, synthetic communities and natural substrates rather than as individual isolates.

MpAPr1, encoding an acid protease from the wine yeast Metschnikowia pulcherrima IWBT Y1123, was previously isolated and shown to display potential activity against casein and grape proteins. However, its characterisation remained partial.; Results: MpAPr1 was cloned into the pGAPZαA vector and transformed into Komagataella pastoris X33 for heterologous expression. After verification of activity, the enzyme properties were characterised. Protease activity within the concentrated supernatant was retained over a pH range of 3.0 to 5.0 and between 10 °C and 50 °C. Optimal conditions for protease activity were found at 40 °C and pH 4.5. Activity was mostly unaffected by the presence of metal ions with the exception of Cu2+ and Ni2+ . Furthermore, proteolytic activity was retained in the presence of sugar and ethanol. pH and temperature conditions for MpAPr1 expression in K. pastoris were optimised. Purification was achieved by means of cation exchange chromatography and kinetic parameters (Km and Vmax ) were determined. MpAPr1 activity against grape proteins was confirmed, but the extent of the degradation was dependent on the nature of these proteins and the environmental conditions.; Conclusion: Overall, the results suggest that MpAPr1 could be applied in food biotechnology processes such as winemaking. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

BACKGROUND: Lipids produced from oleaginous yeasts are a promising alternative to terrestrial oils. Despite promising cellular yields of lipid however, an industrial process remains elusive. One key processing bottleneck is the need to provide nutrient-rich conditions for cellular growth and then extended nutrient-depleted conditions for lipid accumulation. Surprisingly, investigations detailing process development, particularly with a focus on kinetics, are rare in this field.RESULTS: In this investigation we report on the unique oleaginous yeast Metschnikowia pulcherrima, where lipid accumulation ≥ 29.8% (w/w) was achieved without apparent nutrient limitation. The process was developed in stirred tank reactors through determining the influence of temperature, pH and nutrition on lipid production. A temperature of up to 25 °C and initial pH 5 could be applied to enhance initial reaction kinetics. Through the increased supply of yeast extract of up to 5% (w/w) of glucose, a maximum lipid production rate of 0.60 g L−1 h−1 (4 h-average), productivity of 0.29 g L−1 h−1, and yield of 0.17 g g−1 glucose were achieved – the highest yet recorded with this yeast. Suitable to combat the excessive secretion of polyols of up to 0.11 g g−1 glucose was an excess nutrient supply as well as a low cultivation temperature of 15 °C and moderate pH 5.CONCLUSIONS: This study demonstrates that the lack of a starvation stage, coupled with effective process development is required for oleaginous yeasts to achieve the yields and productivities required for commercial lipid production.This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 665992

During the last decade, the use of innovative yeast cultures of both Saccharomyces cerevisiae and non-Saccharomyces yeasts as alternative tools to manage the winemaking process have turned the oenology industry. Although the contribution of different yeast species to wine quality during fermentation is increasingly understood, information about their role in wine ageing over lees is really scarce. This work aims to analyse the incidence of three non-Saccharomyces yeast species of oenological interest (Torulaspora delbrueckii, Lachancea thermotolerans and Metschnikowia pulcherrima) and of a commercial mannoprotein-overproducer S. cerevisiae strain compared with a conventional industrial yeast strain during wine ageing over lees. To evaluate their incidence in mouthfeel properties of wine after 4 months of ageing, the mannoprotein content of wines was evaluated, together with other wine analytic parameters, such as colour and aroma, biogenic amines and amino acids profile. Some differences among the studied parameters were observed during the study, especially regarding the mannoprotein concentration of wines. Our results suggest that the use of T. delbrueckii lees in wine ageing is a useful tool for the improvement of overall wine quality by notably increasing mannoproteins, reaching values higher than obtained using a S. cerevisiae overproducer strain. Copyright © 2016 John Wiley & Sons, Ltd.

Soybean residue (okara), a by-product of soymilk, is produced in large volumes by the soy food industry and is often discarded due to its undesirable flavour. As it contains a considerable amount of protein and fats, biotransformation of okara to improve its flavour presents an opportunity for alternative utilisation. This paper evaluated 10 yeasts in the solid-state fermentation of okara based on their volatile profiles as analysed with HS-SPME GC-MS/FID. Four ?dairy yeasts? (Geotrichum candidum, Yarrowia lipolytica, Debaryomyces hansenii and Kluyveromyces lactis) and six ?wine yeasts? (Saccharomyces cerevisiae, Lachancea thermotolerans, Metschnikowia pulcherrima, Pichia kluyveri, Torulaspora delbrueckii, and Williopsis saturnus) were studied. Results The main off-odourants in okara, hexanal and trans-2-hexenal, significantly decreased after fermentation due to their bioconversion into methyl ketones and/or esters. The okara fermented by dairy yeasts contained greater proportions of methyl ketones, while that by wine yeasts contained more ethyl and acetyl esters. Notably, the okara fermented by W. saturnus contained 13 esters and the total GC-FID peak area of esters was about 380 times that in fresh okara, leading to a perceptible fruity note. Conclusion Okara can be exploited as an inexpensive substrate for bioflavour extraction and/or a more pleasant food ingredient via yeast fermentation. ? 2016 Society of Chemical Industry

The potential of using an antagonistic yeast alone or in combination with microwave treatment for controlling blue mould rot of jujube fruit, and its effect on postharvest quality of fruit, was investigated. The results showed that the growth of Penicillium citrinum was completely inhibited by a 2450-MHz microwave heating for 2 or more minutes in vitro. The population density of P. citrinum in surface wounds of fruit treated with microwave treatment for 2–3 min was significantly lower than that of controls. When tested on jujube fruit, antagonistic yeast or microwave treatment, as stand-alone treatment, the disease incidence of infected wounds was reduced from 100% to 45.0% and 36.0%, and lesion diameters were reduced from 1.92 cm to 1.50 cm and 1.38 cm, respectively. However, in fruit treated with a combination of Metschnikowia pulcherrima and microwave treatment, the disease incidence of infected wounds and lesion diameters was only 21.0% and 1.00 cm, respectively. The natural decay incidence on jujube fruit treated with the combination of microwave treatment and M. pulcherrima was 6.2% after storage at 2 ± 1°C for 45 days and at 22°C for 7 days. None of the treatments impaired quality parameters of fruits. Thus, the combination of microwave treatment and M. pulcherrima could provide an alternative to synthetic fungicides for controlling postharvest blue mould rot of jujube fruit.

UNLABELLED The aim of this work was to determine the chemical constituents and in vitro antimicrobial activity of the essential oil (EO) of the aerial parts of Mentha sueveolens spp. insularis grown in Sardinia (Italy) against probiotic and starter microorganisms. The gas chromatography-mass spectrometry (GC-MS) analysis allowed to identified 34 compounds, most of oxygenated monoterpene compounds (82.5%) and among them, pulegone was found as major compound (46.5%). The agar diffusion test carried out employing the EO of Mentha suaveolens spp. insularis showed a low antibacterial activity, in particular no action was noticed for probiotic bacteria belonging to lactic acid bacteria groups, whereas almost all yeasts strains tested were inhibited. The automated microtitter dilution assay showed a clear effect at increasing concentration of EO on the specific growth rate (μ) and extension of the lag phase (λ) only for S. xylosus SA23 among bacteria and for Saccharomyces cerevisiae, Tetrapisispora phaffii CBS 4417, Metschnikowia pulcherrima, and Candida zemplinina among yeasts. Results obtained in this work allow us to broaden the knowledge on the effect of EOs on probiotic and food-related microorganisms. PRACTICAL APPLICATION Mentha suaveolens spp. insularis may be used in combination with probiotic bacteria into the food matrix or encapsulated in coating and edible films for food preservation.

Zusammenfassung: Es wird fiber den Nachweis von Candida pulcherrima (Lindner) Windisch aus 3 Blutkulturen sowie aus der dem Broviac-Katheter abgesaugten FlUssigkeit eines Patienten mit ambulanter parenteraler Ernahrung, als Ursache plotzlich auftretender Fieberschttbe berichtet. Diese imperfekte Hefeart, die ihren natUrlichen Standort auf Blumen, Obst und Insekten hat, wurde bisher im Zusanunenhang mit einer Erkrankung beim Menschen noch nicht beschrieben. Summary: Candida pulcherrima (Lindner) Windisch was isolated from 3 different blood cultures and the fluid of the Broviac-catheter as the causative agent for recurrent fever in a patient with home parenteral nutrition. This imperfect yeast, which naturally occurs on flowers, fruits or insects has hitherto not been described to cause human infections.

In this study, an integrated approach was evaluated for the control of postharvest decays of mandarin including some pre- and post-harvest treatments under storage conditions. The efficacy of the treatments both as alone and in combination was evaluated during 3 years. Preharvest application of benomyl resulted in significantly less decay of mandarin fruit after storage in 3-year tests. Calcium chloride (CaCl 2 ), 2,4-dichlorophenoxyacetic acid (2,4-D) and gibberellic acid (GA 3 ) as stand-alone treatments or combinations were not effective in controlling Penicillium and total decay infections on inoculated mandarin. Postharvest application of imazalil (200 μg/ml) in solution heated to 54°C for controlling postharvest green and total decay of mandarin was significantly effective for 3 months under storage conditions. The biocontrol activity of yeast (Metschnikowia pulcherrima) was improved when yeast treatment was combined with imazalil (200 μg/ml) at postharvest. The data suggest that preharvest application of benomyl and postharvest treatments of imazalil, hot water and yeast may reduce postharvest green mould and total decay of mandarin under storage conditions.

One hundred and eleven yeast strains were isolated from 60 agricultural soil samples. The samples were taken from four various fields located in the southwest of Slovakia. Cryptococcus laurentii, Candida maltosa, Metschnikowia pulcherrima, and Sporobolomyces salmonicolor were the predominant species in the samples collected from all four types of fields. These species represented 78.4-86.6% of the total yeast counts.The results obtained enabled comparisons to be made between forest and agricultural soil yeast population. We have found out that the yeast population in tilled soils was significantly reduced. The number of yeasts in the tilled soils ranged from 40 to 6.8 x 10(3) CFU/g soil and the average number reached approximately 1.12 x 10(3). This number is more than ten times lower in comparison with the forest soils.

The diversity of yeast species and strains was monitored by physiological tests and a simplified method of karyotyping of yeast chromosomes. During the first phase of investigated alcoholic fermentations, the yeast species Metschnikowia pulcherrima and Hanseniaspora uvarum were predominant, irrespective of the origin of the grape must. At the beginning of fermentation H. uvarum was even present in the case of induced fermentations with dried yeast. Middle and end phase of the alcoholic fermentation were clearly dominated by the yeast species Saccharomyces cerevisiae. In the case of spontaneous fermentations, several different strains of S. cerevisiae were present and competed with each other, whereas in induced fermentations only the inoculated strain of S. cerevisiae was observed. A competition of strains of S. cerevisiae also occurred during the fermentation with dried yeast product consisting of two different strains. An effect of H. uvarum on taste and flavour of wines can be postulated according to the frequency of its appearance during the first phase of fermentation. With the method of rapid karyotyping and supplementary physiological tests it was possible to make reliable assertions about the yeast diversity during alcoholic fermentation.

A patient with short bowel syndrome as a consequence of multiple intestinal resections for Crohn's disease, had a port system implanted to improve her nutritional status. One year later she presented with fever, weakness and nighttime sweating. Metschnikowia pulcherrima Pitt et Miller was grown in blood cultures from the port system. After antifungal chemotherapy using fluconazole and removal of the implant, the patient's condition improved markedly and her fever and sweating disappeared. We conclude that Metschnikowia pulcherrima can turn into a human pathogen in patients with indwelling catheters for parenteral nutrition. Chemotherapy with fluconazole and, whenever possible, removal of the implant, appear to be adequate treatment.

A bacterium capable to lyse viable yeast cells was isolated from compost enriched with baker's yeast cells and was identified as Arthrobacter globiformis. The yeast lytic enzyme complex produced by shaking culture was precipitated with ammonium sulfate, dialyzed and lyophilized. The crude residue contained beta-glucanase and alpha-mannanase, yet not chitinase. Optimale carbon sources in the culture medium for a high enzyme synthesis were 0.5% beta-glucan for the production of beta-glucanase, resp., 3% alpha-mannan for the production of alpha-mannanase. Addition of 10% whole died baker's yeast cells to the culture medium effected similar results. The crude enzyme residue released among other things spheroplasts from cells of the yeast Pichia membranaefaciens, Metschnikowia pulcherrima, Hansenula anomala and Candida guilliermondii "H". However, it did not or only weakly lyse viable cells of the yeasts Rhodotorula rubra, Rhodosporidium toruloides and Sporobolomyces roseus. The mutants of Candida guilliermondii "H" with modified glucan, resp., mannan concentrations in the cell walls did not indicate differences in their susceptibility to lysis.

A bacterium capable to lyse viable yeast cells was isolated from compost enriched with baker's yeast cells and was identified as Arthrobacter globiformis. The yeast lytic enzyme complex produced by shaking culture was precipitated with ammonium sulfate, dialyzed and lyophilized. The crude residue contained β-glucanase and α-mannanase, yet not chitinase. Optimale carbon sources in the culture medium for a high enzyme synthesis were 0.5% β-glucan for the production of β-glucanase, resp., 3% α-mannan for the production of α-mannanase. Addition of 10% whole died baker's yeast cells to the culture medium effected similar results. The crude enzyme residue released among other things spheroplasts from cells of the yeast Pichia membranaefaciens, Metschnikowia pulcherrima, Hansenula anomala and Candida guilliermondii “H”. However, it did not or only weakly lyse viable cells of the yeasts Rhodotorula rubra, Rhodosporidium toruloides and Sporobolomyces roseus. The mutants of Candida guilliermondii “H” with modified glucan, resp., mannan concentrations in the cell walls did not indicate differences in their susceptibility to lysis.

The effects of temperature and pH on the survival and growth of Saccharomyces cerevisiae, Kloeckera apiculata, Candida stellata, Candida krusei, Candida pulcherrima and Hansenula anomala were examined during mixed culture in grape juice. At 25°C, pH 3.0 and pH 3.5, S. cerevisiae dominated the fermentation and the other species died off before fermentation was completed. Saccharomyces cerevisiae also dominated the fermentation at 20°C but there was increased growth and survival of the other species. At 10°C the fermentation was dominated by the growth of both S. cerevisiae and K. apiculata and there was extended growth and survival of C. stellata and C. krusei. Juices fermented at 10°C exhibited ethanol concentrations between 7.4 and 13.4% and populations of K. apiculata, C. stellata and C. krusei in the range 106-108 cells/ml. However, these species produced maximum ethanol concentrations in the range 2.7–6.6% when grown as single cultures in grape juice.

Six morphologically different groups of yeasts comprising Kloeckera apiculata. Candida krusei, Candida valida, Candida sorbosa, Metschnikowia pulcherrima and Saccharomyces chevalieri were isolated from fresh, fermenting and fermented apple juice. This is the first report on the isolation of C. valida, C. sorbosa and S. chevalieri from fermenting and fermented apple juice.