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BMC Microbiology Nov 2023Plant fungal pathogens cause substantial economic losses through crop yield reduction and post-harvest storage losses. The utilization of biocontrol agents presents a...
BACKGROUND
Plant fungal pathogens cause substantial economic losses through crop yield reduction and post-harvest storage losses. The utilization of biocontrol agents presents a sustainable strategy to manage plant diseases, reducing the reliance on hazardous chemical. Recently, Pichia kudriavzevii has emerged as a promising biocontrol agent because of its capacity to inhibit fungal growth, offering a potential solution for plant disease management.
RESULTS
Two novel Pichia kudriavzevii strains, Pk_EgyACGEB_O1 and Pk_EgyACGEB_O2, were isolated from olive brine samples. The microscopic characterization of the strains revealed similar structures. However, there were noticeable differences in their visual morphology. Based on their internal transcribed spacer (ITS) DNA sequences, Pk_EgyACGEB_O1 and Pk_EgyACGEB_O2 strains assigned by GenBank IDs MZ507552.1 and MZ507554.1 shared high sequence similarity (~ 99.8% and 99.5%) with P. kudriavzevii, respectively. Both strains were evaluated in vitro against plant pathogenic fungi. The strains revealed the ability to consistently inhibit fungal growth, with Pk_EgyACGEB_O2 showing higher effectiveness. In addition, both P. kudriavzevii strains effectively controlled grey mold disease caused by B. cinerea in golden delicious apples, suggesting their potential as sustainable and eco-friendly biocontrol agents for post-harvest diseases. Based on a comprehensive bioinformatics pipeline, candidate-secreted proteins responsible for the potent antifungal activity of P. kudriavzevii were identified. A total of 59 proteins were identified as common among the P. kudriavzevii CBS573, SD108, and SD129 strains. Approximately 23% of the secreted proteins in the P. kudriavzevii predicted secretome are hydrolases with various activities, including proteases, lipases, glycosidases, phosphatases, esterases, carboxypeptidases, or peptidases. In addition, a set of cell-wall-related proteins was identified, which might enhance the biocontrol activity of P. kudriavzevii by preserving the structure and integrity of the cell wall. A papain inhibitor was also identified and could potentially offer a supplementary defense against plant pathogens.
CONCLUSION
Our results revealed the biocontrol capabilities of P. kudriavzevii against plant pathogenic fungi. The research focused on screening novel strains for their ability to inhibit the growth of common pathogens, both in vitro and in vivo. This study shed light on how P. kudriavzevii interacts with fungal pathogens. The findings can help develop effective strategies for managing plant diseases.
Topics: Pichia; Antifungal Agents; Mycoses; Plant Diseases
PubMed: 37980509
DOI: 10.1186/s12866-023-03047-w -
AMB Express May 2018The yeast strain SJP-SNU was investigated as a probiotic and was characterized with respect to growth temperature, bile salt resistance, hydrogen sulfide reducing...
The yeast strain SJP-SNU was investigated as a probiotic and was characterized with respect to growth temperature, bile salt resistance, hydrogen sulfide reducing activity, intestinal survival ability and chicken embryo pathogenicity. In addition, we determined the complete genomic and mitochondrial sequences of SJP-SNU and conducted comparative genomics analyses. SJP-SNU grew rapidly at 37 °C and formed colonies on MacConkey agar containing bile salt. SJP-SNU reduced hydrogen sulfide produced by Salmonella serotype Enteritidis and, after being fed to 4-week-old chickens, could be isolated from cecal feces. SJP-SNU did not cause mortality in 10-day-old chicken embryos. From 13 initial contigs, 11 were finally assembled and represented 10 chromosomal sequences and 1 mitochondrial DNA sequence. Comparative genomic analyses revealed that SJP-SNU was a strain of Pichia kudriavzevii. Although SJP-SNU possesses pathogenicity-related genes, they showed very low amino acid sequence identities to those of Candida albicans. Furthermore, SJP-SNU possessed useful genes, such as phytases and cellulase. Thus, SJP-SNU is a useful yeast possessing the basic traits of a probiotic, and further studies to demonstrate its efficacy as a probiotic in the future may be warranted.
PubMed: 29774473
DOI: 10.1186/s13568-018-0609-0 -
Food Chemistry: X Jun 2023Mixed fermentation using saccharomyces cerevisiae and non- has become one of the main research strategies to improve wine aroma. Hence, this study applied the mixed...
Mixed fermentation using saccharomyces cerevisiae and non- has become one of the main research strategies to improve wine aroma. Hence, this study applied the mixed fermentation technique using and to brew Cabernet Sauvignon wine and to investigate the effects of inoculation timing and inoculation ratio on the polyphenolics, antioxidant activity and aroma of the resulting wine. The results showed that mixed fermentation significantly improved the amounts of flavan-3-ols. In particular, S1:5 had the highest amounts of (-)-catechin and procyanidin B1 (73.23 mg/L and 46.59 mg/L), while S1:10 had the highest (-)-epicatechin content (57.95 mg/L). Meanwhile, S1:10 showed the strongest FRAP, CUPRAC and ABTS + activities (31.46 %, 25.38 % and 13.87 % higher than that of CK, respectively). In addition, mixed fermentation also increased the amounts of phenylethanol, isoamyl alcohol and ethyl esters, which enhanced the rose-like and fruity flavor of wine. This work used a friendly non- alongside appropriate inoculation strategies to provide an alternative approach for improved wine aroma and phenolic profile.
PubMed: 37131849
DOI: 10.1016/j.fochx.2023.100685 -
Microbiology Spectrum Dec 2021This study investigated the effect of Ca ascorbate on the biocontrol efficacy of Pichia kudriavzevii and the possible mechanisms. The results indicated that the...
This study investigated the effect of Ca ascorbate on the biocontrol efficacy of Pichia kudriavzevii and the possible mechanisms. The results indicated that the biocontrol activity of was significantly enhanced by 0.15 g L of Ca ascorbate, with higher growth rates of yeast cells and . The antioxidant enzyme activity in , including catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD), were improved by Ca ascorbate and reached the maximum at 96 h, 96 h, and 72 h, respectively. The expression of the antioxidant enzyme-related genes (8.55-fold) and (7.26-fold) peaked at 96 h, while (2.8-fold) peaked at 48 h, which were similar to the trends of enzyme activities. Compared with the control, 0.15 g L of Ca ascorbate and CaCl increased the activity of succinate dehydrogenase in , thereby enhancing the utilization of nutrients by yeast cells, and calcium ascorbate had the strongest effect. The expressions of , , , and were significantly higher in the Ca ascorbate treatment than the other groups, and the CaCl treatment was also significantly higher than the control. These results indicated that Ca ascorbate can effectively improve the energy metabolism and cell wall synthesis and slow down the senescence of yeast cells. In general, Ca ascorbate can improve the environmental adaptability of and thus improve the biocontrol effect, which is associated with inducing antioxidant enzymes in yeast cells and enhancing energy metabolism and nutrient utilization efficiency to increase nutrient competition with pathogens. Antagonistic yeast is a promising way to control postharvest fruit decay because of its safety and broad-spectrum resistance. However, the biocontrol efficacy of yeast is limited by environmental stress, such as oxidative stress. Therefore, the improvement of antioxidant capacity has become a research hot spot in improving the biocontrol efficacy of yeast. The induction of Ca ascorbate on the antioxidant capacity and physiological activity of yeast was studied. The results showed better induction of antioxidant enzyme and physiological activity in yeast by Ca ascorbate for better antioxidant capacity, and Ca also played a synergistic promotion effect, which improved the biocontrol efficacy. These results provide an approach for the research and application of improving the environmental adaptability and biocontrol effectiveness of yeast.
Topics: Antibiosis; Antioxidants; Ascorbic Acid; Biological Control Agents; Botrytis; Catalase; Fruit; Solanum lycopersicum; Oxidative Stress; Pichia; Plant Diseases
PubMed: 34937188
DOI: 10.1128/spectrum.01507-21 -
Frontiers in Microbiology 2021and were used as starter cultures to conduct inoculated wet fermentations of coffee beans, and their growth, metabolic activities and impact on the flavor, aroma and...
and were used as starter cultures to conduct inoculated wet fermentations of coffee beans, and their growth, metabolic activities and impact on the flavor, aroma and overall sensory quality of coffee were compared with spontaneous fermentation (control). and dominated the fermentations, growing to maximum populations of about 10.0 log CFU/ml compared with 8.0 log CFU/ml in the spontaneous fermentation. The dominance of the inoculated yeasts led to faster and more complete utilization of sugars in the mucilage, with resultant production of 2-3 fold higher concentrations of metabolites such as glycerol, alcohols, aldehydes, esters, and organic acids in the fermented green beans. Cup tests showed coffee produced from the inoculated fermentations, especially with , received higher scores for flavor, aroma and acidity than the control. The findings of this study confirmed the crucial role of yeasts in the wet fermentation of coffee beans and their contribution to high quality coffee, and demonstrated the potential and as starter cultures in the process.
PubMed: 34421873
DOI: 10.3389/fmicb.2021.713969 -
Journal, Genetic Engineering &... Sep 2022Fermented foods are the results of metabolic activities of various microorganisms. People have traditionally known how to culture desirable microorganisms, primarily...
BACKGROUND
Fermented foods are the results of metabolic activities of various microorganisms. People have traditionally known how to culture desirable microorganisms, primarily lactic acid bacteria, yeasts, and filamentous molds, for the manufacture of edible foods. Yeast isolated from home-made mango pickle from Hamirpur, Himachal Pradesh, was assessed for probiotic properties and their enzymatic profiling.
RESULTS
Four yeast isolates were isolated out of which P. kudriavzevii Y33 was selected on the basis of high acid tolerance as well as broadest antimicrobial activity. The selected isolate was observed to have high acid tolerance at pH 2 and show strong antimicrobial activity against all the pathogens examined. P. kudriavzevii Y33 can also withstand high bile concentration and showed high viability index, i.e., 95% at concentration of 2% of bile. The isolate was able to demonstrate high cholesterol assimilation in medium containing ox bile and taurocholate, at 88.58 and 86.83%, respectively. The autoaggregation ability of isolate increases with increasing the time of incubation and showed 87% of autoaggregation after 24 h of incubation. P. kudriavzevii Y33 exhibited resistance towards different antibiotics, found to be positive for exopolysaccharide production and showed no hemolytic activity. The isolate was observed to produce several enzymes such as β-galactosidase, protease, amylase, phytase, and lipase.
CONCLUSIONS
The results of the current study revealed that P. kudriavzevii Y33 has various beneficial qualities that suggest it could be used as probiotics. Enzymes produced by yeast isolate help in improving flavor and mineral availability in the fermented products.
PubMed: 36083419
DOI: 10.1186/s43141-022-00416-2 -
Biotechnology For Biofuels Nov 2021High-temperature bioethanol production benefits from yeast thermotolerance. Salt stress could induce obvious cross-protection against heat stress of Pichia kudriavzevii,...
Salt stress improves thermotolerance and high-temperature bioethanol production of multi-stress-tolerant Pichia kudriavzevii by stimulating intracellular metabolism and inhibiting oxidative damage.
BACKGROUND
High-temperature bioethanol production benefits from yeast thermotolerance. Salt stress could induce obvious cross-protection against heat stress of Pichia kudriavzevii, contributing to the improvement of its thermotolerance and bioethanol fermentation. However, the underlying mechanisms of the cross-protection remain poorly understood.
RESULTS
Salt stress showed obvious cross-protection for thermotolerance and high-temperature ethanol production of P. kudriavzevii observed by biomass, cell morphology and bioethanol production capacity. The biomass and ethanol production of P. kudriavzevii at 45 °C were, respectively, improved by 2.6 and 3.9 times by 300 mmol/L NaCl. Metabolic network map showed that salt stress obviously improved the key enzymes and intermediates in carbohydrate metabolism, contributing to the synthesis of bioethanol, ATP, amino acids, nucleotides, and unsaturated fatty acids, as well as subsequent intracellular metabolisms. The increasing trehalose, glycerol, HSPs, and ergosterol helped maintain the normal function of cell components. Heat stress induced serious oxidative stress that the ROS-positive cell rate and dead cell rate, respectively, rose from 0.5% and 2.4% to 28.2% and 69.2%, with the incubation temperature increasing from 30 to 45 °C. The heat-induced ROS outburst, oxidative damage, and cell death were obviously inhibited by salt stress, especially the dead cell rate which fell to only 20.3% at 300 mmol/L NaCl. The inhibiting oxidative damage mainly resulted from the abundant synthesis of GSH and GST, which, respectively, increased by 4.8 and 76.1 times after addition of 300 mmol/L NaCl. The improved bioethanol production was not only due to the improved thermotolerance, but resulted from the up-regulated alcohol dehydrogenases and down-regulated aldehyde dehydrogenases by salt stress.
CONCLUSION
The results provide a first insight into the mechanisms of the improved thermotolerance and high-temperature bioethanol production of P. kudriavzevii by salt stress, and provide important information to construct genetic engineering yeasts for high-temperature bioethanol production.
PubMed: 34823567
DOI: 10.1186/s13068-021-02071-0 -
Biotechnology For Biofuels 2020Although bioethanol production has been gaining worldwide attention as an alternative to fossil fuel, ethanol productivities and yields are still limited due to the...
BACKGROUND
Although bioethanol production has been gaining worldwide attention as an alternative to fossil fuel, ethanol productivities and yields are still limited due to the susceptibility of fermentation microorganisms to various stress and inhibitory substances. There is therefore an unmet need to search for multi-stress-tolerant organisms to improve ethanol productivity and reduce production cost, particularly when lignocellulosic hydrolysates are used as the feedstock.
RESULTS
Here, we have characterized a previously isolated LC375240 strain which is thermotolerant to high temperatures of 37 °C and 42 °C. More excitingly, growth and ethanol productivity of this strain exhibit strong tolerance to multiple stresses such as acetic acid, furfural, formic acid, HO and high concentration of ethanol at 42 °C. In addition, simple immobilization of LC375240 on corncobs resulted to a more stable and higher efficient ethanol production for successive four cycles of repeated batch fermentation at 42 °C.
CONCLUSION
The feature of being thermotolerant and multi-stress-tolerant is unique to LC375240 and makes it a good candidate for second-generation bioethanol fermentation as well as for investigating the molecular basis underlying the robust stress tolerance. Immobilization of LC375240 on corncobs is another option for cheap and high ethanol productivity.
PubMed: 32477425
DOI: 10.1186/s13068-020-01729-5 -
Mycopathologia Aug 2022Revealing the phylogenetic relationships of Candida krusei strains (sexual form Pichia kudriavzevii) is a prerequisite for understanding the evolution of its...
Revealing the phylogenetic relationships of Candida krusei strains (sexual form Pichia kudriavzevii) is a prerequisite for understanding the evolution of its virulence-associated mechanisms and ecological lifestyles. Molecular phylogenetic analyses based on entire internal transcribed spacer region (ITS) and multilocus sequence typing (MLST) data were carried out with sequences available in public databases and Hungarian isolates from animals obtained for the study. The ITS haplotype network yielded a high frequency haplotype at the centre of the network (H1; n = 204) indicating that various selective pressure might resulted in population expansion from H1. MLST analysis identified three new genotypes among animal-derived isolates, therefore overall 203 sequence types were investigated to determine the population structure of C. krusei. The most commonly encountered sequence types were ST 17 and ST 67. Phylogenetic analyses showed diverse genetic construction of C. krusei population. Evidence of potential recombination events were also observed that might play some role in high intraspecies genetic variability among strains, however, the limited data of C. krusei genotypes from different countries prevented us to identify accurate evolutionary routes of commensal and pathogenic strains or species-specific lineages. Further expansion of C. krusei MLST database may promote the better understanding of the mixed evolutionary history of this species.
Topics: Candida; Multilocus Sequence Typing; Phylogeny; Pichia
PubMed: 35689765
DOI: 10.1007/s11046-022-00640-x -
Journal of Fungi (Basel, Switzerland) Dec 2021Improving the comprehensive utilization of sugars in lignocellulosic biomass is a major challenge for enhancing the economic viability of lignocellulose biorefinement. A...
Improving the comprehensive utilization of sugars in lignocellulosic biomass is a major challenge for enhancing the economic viability of lignocellulose biorefinement. A robust yeast N-X showed excellent performance in ethanol production under high temperature and low pH conditions and was engineered for ᴅ-xylonate production without xylitol generation. The recombinant strain N-X/S1 was employed for sequential production of ᴅ-xylonate and ethanol from ᴅ-xylose, feeding on ᴅ-glucose without pH control in a two-stage strategy of aerobic and shifting micro-aerobic fermentation. Acid-pretreated corncob without detoxification and filtration was used for ᴅ-xylonate production, then simultaneous saccharification and ethanol fermentation was performed with cellulase added at pH 4.0 and at 40 °C. By this strategy, 33.5 g/L ᴅ-xylonate and 20.8 g/L ethanol were produced at yields of 1.10 g/g ᴅ-xylose and 84.3% of theoretical value, respectively. We propose a promising approach for the sequential production of ᴅ-xylonate and ethanol from non-detoxified corncob using a single microorganism.
PubMed: 34947020
DOI: 10.3390/jof7121038