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Food Chemistry Apr 2024This study is the first to demonstrate the yeast Pichia kudriavzevii can effectively deliver Se and investigate the distribution and species of Se in Se-enriched P....
This study is the first to demonstrate the yeast Pichia kudriavzevii can effectively deliver Se and investigate the distribution and species of Se in Se-enriched P. kudriavzevii. Results showed that P. kudriavzevii can accumulate Se and convert 84.883% of absorbed Se into organic forms, of which 78.338% was incorporated into protein, 1.978% combined with polysaccharides, and 0.456% bound to nucleic acid. Besides, water-soluble, salt-soluble, and alkali-soluble proteins account for 49.398%, 1.867%, and 20.628% of selenoprotein, respectively. The dominant Se species were SeCys2 and MeSeCys. Additionally, Se-enrichment enhanced nutritional value of P. kudriavzevii by increasing the levels of amino acids, iron, and zinc. The activity of key rate-limiting enzyme sephosphate synthetase involved in Se biotransformation was improved after Se enrichment. The extracellular pH results suggest that Se enrichment ability can be further enhanced by elevating pH. These results suggest P. kudriavzevii holds great promise as an effective vehicle for delivering Se.
Topics: Selenium; Pichia; Biotransformation; Zinc
PubMed: 37976881
DOI: 10.1016/j.foodchem.2023.137966 -
Scientific Reports Nov 2023This study addresses the environmental risks associated with the accumulation of keratin waste from poultry, which is resistant to conventional protein degradation...
This study addresses the environmental risks associated with the accumulation of keratin waste from poultry, which is resistant to conventional protein degradation methods. To tackle this issue, microbial keratinases have emerged as promising tools for transforming resilient keratin materials into valuable products. We focus on the Metalloprotease (MetPr) gene isolated from novel Pichia kudriavzevii YK46, sequenced, and deposited in the NCBI GenBank database with the accession number OQ511281. The MetPr gene encodes a protein consisting of 557 amino acids and demonstrates a keratinase activity of 164.04 U/ml. The 3D structure of the protein was validated using Ramachandran's plot, revealing that 93% and 97.26% of the 557 residues were situated within the most favoured region for the MetPr proteins of template Pichia kudriavzevii strain 129 and Pichia kudriavzevii YK46, respectively. Computational analyses were employed to determine the binding affinities between the deduced protein and beta keratin. Molecular docking studies elucidated the optimal binding affinities between the metalloprotease (MetPr) and beta-keratin, yielding values of - 260.75 kcal/mol and - 257.02 kcal/mol for the template strains Pichia kudriavzevii strain 129 and Pichia kudriavzevii YK46, respectively. Subsequent molecular cloning and expression of the MetPr gene in E. coli DH5α led to a significantly higher keratinase activity of 281 ± 12.34 U/ml. These findings provide valuable insights into the potential of the MetPr gene and its encoded protein for keratin waste biotransformation, with implications for addressing environmental concerns related to keratinous waste accumulation.
Topics: Animals; Feathers; Escherichia coli; Molecular Docking Simulation; Pichia; Metalloproteases; Keratins; Cloning, Molecular
PubMed: 37968282
DOI: 10.1038/s41598-023-47179-5 -
Current Opinion in Critical Care Oct 2023The increasing incidence of drug-resistant Candida brings a new challenge to the treatment of invasive candidiasis. Although cross-resistance among azoles and... (Review)
Review
PURPOSE OF REVIEW
The increasing incidence of drug-resistant Candida brings a new challenge to the treatment of invasive candidiasis. Although cross-resistance among azoles and echinocandins was generally uncommon, reports of multidrug-resistant (MDR) Candida markedly increased in the last decade. The purpose of this review is to understand mechanisms and risk factors for resistance and how to tackle antifungal resistance.
RECENT FINDINGS
The paper describes the action of the three main classes of antifungals - azoles, echinocandins and polyenes - and Candida's mechanisms of resistance. The current evolution from cross-resistance to multiresistance among Candida explains the modern glossary - multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR) - imported from bacteria. MDR Candida most commonly involves acquired resistance in species with intrinsic resistance, therefore it mostly involves C. glabrata, C. parapsilosis, C. krusei, C guilliermondii or C. auris , which is intrinsically multidrug resistant. Finally, strategies to tackle antifungal resistance became clearer, ideally implemented through antifungal stewardship.
SUMMARY
Avoiding antifungal's overuse and selecting the best drug, dose and duration, when they are needed, is fundamental. Knowledge of risk factors for resistance, microbiological diagnosis to the species, use of susceptibility test supported by antifungal stewardship programs help attaining effective therapy and sustaining the effectiveness of the current antifungal armamentarium.
Topics: Humans; Candida; Antifungal Agents; Candidiasis, Invasive; Echinocandins; Azoles
PubMed: 37641511
DOI: 10.1097/MCC.0000000000001077 -
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 -
Letters in Applied Microbiology Sep 2023Film-forming yeasts are potential sources of defects in alcoholic beverages. The aim of this study is to assess the growth capacity of Pichia and Candida film-forming...
Film-forming yeasts are potential sources of defects in alcoholic beverages. The aim of this study is to assess the growth capacity of Pichia and Candida film-forming yeasts in cider and wine and the effects on their chemical composition. Cider, partially and fully fermented wine were inoculated with strains of C. californica, P. fermentans, P. kluyveri, P. kudriavzevii, P. manshurica, and P. membranifaciens to simulate a post-fermentative contamination. The former three species grew only in cider. Pichia manshurica and P. kudriavzevii displayed high viability in wine up to 13.18% (v v-1) ethanol. Significant changes in odour-active molecules from different chemical groups were observed in cider and wine in the inoculated samples, compared to the non-inoculated ones. Cider is more susceptible to contamination by all of the species tested, due to its low alcohol content, while P. membranifaciens, P. manshurica, and P. kudriavzevii are additionally potential spoilage agents of wine. This study highlights the risk of cider and wine contamination by film-forming yeasts. Their impact on aroma profiles depends on their ability to grow and their metabolism. This study contributes to an understanding of the possible physiological and metabolic mechanisms responsible for film formation and chemical changes in alcoholic beverages.
Topics: Wine; Fermentation; Pichia; Alcoholic Beverages; Candida
PubMed: 37656878
DOI: 10.1093/lambio/ovad099 -
Brazilian Journal of Microbiology :... Mar 2024Enzymatic compounds can be found abundantly and provide numerous advantages in microbial organisms. Xylanases are used in various pharmaceutical, food, livestock,...
Enzymatic compounds can be found abundantly and provide numerous advantages in microbial organisms. Xylanases are used in various pharmaceutical, food, livestock, poultry, and paper industries. This study aimed to investigate xylanase-producing yeasts, xylose concentration curve and their enzymatic activity under various factors including carbon and nitrogen sources, temperature, and pH. Enzyme activity was evaluated under different conditions before, during, and after purification. The yeast strains were obtained from the wood product workshop and were subsequently cultivated on YPD (yeast extract peptone dextrose) medium. Additionally, the growth curve of the yeast and its molecular identification were conducted. The optimization and design process of xylan isolated from corn wood involved the use of Taguchi software to test different parameters like carbon and nitrogen sources, temperature, and pH, with the goal of determining the most optimal conditions for enzyme production. In addition, the Taguchi method was utilized to conduct a multifactorial optimization of xylanase enzyme activity. The isolated species were partially purified using ammonium sulfate precipitation and dialysis bag techniques. The results indicated that 3 species (8S, 18S, and 16W) after molecular identification based on 18S rRNA gene sequencing were identified as Candida tropicalis SBN-IAUF-1, Candida tropicalis SBN-IAUF-3, and Pichia kudriavzevii SBN-IAUF-2, respectively. The optimal parameters for wheat carbon source and peptone nitrogen source were found at 50 °C and pH 9.0 through single-factor optimization. By using the Taguchi approach, the best combination for highest activity was rice-derived carbon source and peptone nitrogen source at 50 °C and pH 6.0. The best conditions for xylanase enzyme production in single-factor optimization of wheat bran were 2135.6 U/mL, peptone 4475.25 U/mL, temperature 50 °C 1868 U/mL, and pH 9.0 2002.4 U/mL. Among the tested yeast, Candida tropicalis strain SBN-IAUF-1 to the access number MZ816946.1 in NCBI was found to be the best xylanase product. The highest ratio of enzyme production at the end of the delayed phase and the beginning of the logarithmic phase was concluded by comparing the growth ratio of 8S, 16W, and 18S yeasts with the level of enzymatic activity. This is the first report on the production of xylan polymer with a relative purity of 80% in Iran. The extracellular xylanases purified from the yeast species of C. tropicalis were introduced as a desirable biocatalyst due to their high enzymatic activity for the degradation of xylan polymers.
Topics: Wood; Xylans; Candida tropicalis; Peptones; Fermentation; Yeasts; Carbon; Nitrogen; Endo-1,4-beta Xylanases; Pichia
PubMed: 37957443
DOI: 10.1007/s42770-023-01171-3 -
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 -
World Journal of Microbiology &... Sep 2023Esters were identified as the primary volatile flavor compounds in Chinese Baijiu, exerting a significant influence on its quality and aroma. This study focused on the...
Esters were identified as the primary volatile flavor compounds in Chinese Baijiu, exerting a significant influence on its quality and aroma. This study focused on the yeast strain Pichia kudriavzevii, renowned for its high capacity to produce esters. Whole genome sequences were annotated and analyzed using the GO, KEGG, KOG, CAZy, and Pfam databases to determine the genetic basis underly the enhanced ester production capacity. Results showed that P. kudriavzevii gene function was concentrated in biosynthetic capacity, metabolic capacity, amino acid translocation capacity, glycoside hydrolysis capacity and transfer capacity. Additionally, acyltransferase and kinase were predicted as active sites contributing to P. kudriavzevii high ester production. We further compared the volatile composition differences between P. kudriavzevii and Saccharomyces cerevisiae through Headspace solid-phase microextraction-gas Chromatography-mass spectrometry (HS-SPME-GC-MS), revealing P. kudriavzevii produced 3.5 times more esters than S. cerevisiae. Overall, our findings suggest that P. kudriavzevii had potential applications in the Baijiu brewing industry.
Topics: Saccharomyces cerevisiae; Pichia; Amino Acids; Esters
PubMed: 37713136
DOI: 10.1007/s11274-023-03743-9 -
Probiotics and Antimicrobial Proteins Jun 2023Potential probiotic yeast strains isolated from fermented food need to meet safe and beneficial conditions for the host's health. The Pichia kudriavzevii YGM091 strain...
Potential probiotic yeast strains isolated from fermented food need to meet safe and beneficial conditions for the host's health. The Pichia kudriavzevii YGM091 strain isolated from fermented goat milk has outstanding probiotic characteristics, including: the high survival percentage in digestive system conditions (reaching up 247.13 ± 0.12 and 145.03 ± 0.06% at pH 3.0 and bile salt 0.5%, respectively); good tolerance to temperature, salt, phenol, ethanol; good surface properties such as high hydrophobicity percentage (> 60%), the high auto-aggregation percentage rate (66.56 ± 1.45% after 45 min of incubation) and the high co-aggregation percentage rate with pathogenic bacteria in a short time (> 40% after 2 h of incubation); biofilm forming after 24 h of incubation on abiotic surfaces; antioxidant activity reached excellent level after only 24 h of incubation (The percentage free radical scavenging and the Trolox equivalent reaching up 79.86 ± 0.70% and 92.09 ± 0.75 µg/mL after 72 h of incubation); extracellular enzymes production protease and cellulase with high activity, amylase and pectinase with moderate activity and non-lipase activity. Simultaneously, the YGM091 strain is the in vitro safety yeast: insensitive to antibiotics and fluconazole, negative for gelatinase, phospholipase, coagulase, and non-hemolysis activities. Furthermore, this strain is in vivo safety yeast with the dosages below 10 CFU/larva in the Galleria mellonella model with over 90% survival larvae and the yeast density reduced to just 10-10 CFU/larva after 72 h post-injection. Research results have demonstrated that the Pichia kudriavzevii YGM091 strain is a safe potential probiotic yeast and could become a candidate probiotic food to be used in the future.
PubMed: 37368223
DOI: 10.1007/s12602-023-10114-1 -
Archives of Microbiology Aug 2023Recently, there has been growing interest in the characterization of native yeasts for their use in production of wines with regional characteristics. This study aimed...
Recently, there has been growing interest in the characterization of native yeasts for their use in production of wines with regional characteristics. This study aimed to investigate Saccharomyces and non-Saccharomyces yeasts present in the spontaneous fermentation of Tannat and Marselan grape musts collected from Concordia (Entre Ríos, Argentina) over 2019, 2020, and 2021 vintages. The evolution of these fermentative processes was carried out by measuring total soluble solids, total acidity, volatile acidity, pH, ethanol concentration, and total carbon content. Isolated Saccharomyces and non-Saccharomyces yeasts were identified based on colony morphology in WL medium, 5.8S-ITS-RFLP analysis, and 26S rDNA D1/D2 gene sequencing. Two hundred and ten yeast colonies were isolated and identified as Pichia kudriavzevii, Saccharomyces cerevisiae, Hanseniaspora uvarum, Metschnikowia pulcherrima, Candida albicans, Candida parapsilosis, Pichia occidentalis, Pichia bruneiensis, Hanseniaspora opuntiae, Issatchenkia terricola, and Hanseniaspora vineae. P. kudriavzevii isolated from all vintages was associated with the spontaneous fermentation of grape musts from the Concordia region.
Topics: Vitis; Fermentation; Yeasts; Wine; Saccharomyces cerevisiae; DNA, Ribosomal
PubMed: 37550458
DOI: 10.1007/s00203-023-03646-1