-
Mycopathologia Dec 2023We performed a retrospective survey of non-Candida albicans candidemia in patients with cancer, including those with solid tumors and those with hematological...
We performed a retrospective survey of non-Candida albicans candidemia in patients with cancer, including those with solid tumors and those with hematological malignancies as well as transplants patients both, solid-organ transplant recipients and hematopoietic stem cell transplant recipients. The study was performed at two healthcare centers in New York City and covered the years 2018-2022. A total of 292 patients (318 isolates) were included in the study. In order of frequency, C. glabrata (38%) was the most common species recovered, followed by C. parapsilosis (19.2%), C. tropicalis (12.6%), C. krusei (10.7%), C. lusitaniae (5.7%), and C. guilliermondii (4.4%). Micafungin was the most common antifungal treatment and 18.5% of patients were on antifungal prophylaxis. The 30-day crude mortality was 40%. 4.5% of patients had more than one non-albicans species detected. In conclusion, this study represents one of the largest surveys of non-albicans species in cancer and transplant patients and provides data on the current epidemiology of these Candida species in this patient population.
Topics: Humans; Antifungal Agents; Transplant Recipients; Retrospective Studies; Microbial Sensitivity Tests; Candida; Candidemia; Candida glabrata; Candida parapsilosis; Candida tropicalis; Neoplasms
PubMed: 37365379
DOI: 10.1007/s11046-023-00765-7 -
Metabolic Engineering Jul 2023The robust nature of the non-conventional yeast Issatchenkia orientalis allows it to grow under highly acidic conditions and therefore, has gained increasing interest in...
The robust nature of the non-conventional yeast Issatchenkia orientalis allows it to grow under highly acidic conditions and therefore, has gained increasing interest in producing organic acids using a variety of carbon sources. Recently, the development of a genetic toolbox for I. orientalis, including an episomal plasmid, characterization of multiple promoters and terminators, and CRISPR-Cas9 tools, has eased the metabolic engineering efforts in I. orientalis. However, multiplex engineering is still hampered by the lack of efficient multicopy integration tools. To facilitate the construction of large, complex metabolic pathways by multiplex CRISPR-Cas9-mediated genome editing, we developed a bioinformatics pipeline to identify and prioritize genome-wide intergenic loci and characterized 47 gRNAs located in 21 intergenic regions. These loci are screened for guide RNA cutting efficiency, integration efficiency of a gene cassette, the resulting cellular fitness, and GFP expression level. We further developed a landing pad system using components from these well-characterized loci, which can aid in the integration of multiple genes using single guide RNA and multiple repair templates of the user's choice. We have demonstrated the use of the landing pad for simultaneous integrations of 2, 3, 4, or 5 genes to the target loci with efficiencies greater than 80%. As a proof of concept, we showed how the production of 5-aminolevulinic acid can be improved by integrating five copies of genes at multiple sites in one step. We have further demonstrated the efficiency of this tool by constructing a metabolic pathway for succinic acid production by integrating five gene expression cassettes using a single guide RNA along with five different repair templates, leading to the production of 9 g/L of succinic acid in batch fermentations. This study demonstrates the effectiveness of a single gRNA-mediated CRISPR platform to build complex metabolic pathways in a non-conventional yeast. This landing pad system will be a valuable tool for the metabolic engineering of I. orientalis.
Topics: CRISPR-Cas Systems; Saccharomyces cerevisiae; Gene Editing; Succinates
PubMed: 37343658
DOI: 10.1016/j.ymben.2023.06.010 -
Journal of Agricultural and Food... Jun 2023Nontraditional yeasts prevalent in tropical agricultural fermentations such as coffee and cocoa are known to contribute to aroma profiles, yet the functional roles and...
Nontraditional yeasts prevalent in tropical agricultural fermentations such as coffee and cocoa are known to contribute to aroma profiles, yet the functional roles and interactions between the associated microbial consortia in a farm fermentation are unclear. Here, boiled green bean extract (GBE) from green coffee beans was developed as a rich screening medium to deconstruct the microbial consortia and their interactions during the fermentation of dried green coffee beans. When cultivated in coculture with on GBE, strain-specific groupings with distinct volatile organic profiles were observed for nontraditional yeasts (e.g., spp., ). Further changes are evident when constructed consortia composed of nontraditional yeast, , and var. were cultured in GBE, and a comparison with abiotically acidified GBE suggests that pH plays a major role in the influence of lactic acid bacteria (LAB) on fermentation aromas. This approach represents a tool for the development of starter culture formulations to create different flavor profiles in coffee fermentation.
Topics: Fermentation; Saccharomyces cerevisiae; Odorants; Yeasts; Cacao; Chocolate
PubMed: 37310147
DOI: 10.1021/acs.jafc.3c01357 -
Journal of Applied Microbiology Jun 2023To investigate the effect of CgMCUR1 on the phenotype of Candida glycerinogenes and Saccharomyces cerevisiae.
AIMS
To investigate the effect of CgMCUR1 on the phenotype of Candida glycerinogenes and Saccharomyces cerevisiae.
METHODS AND RESULTS
Inhibition of CgMCUR1 expression reduced acetate, H2O2, and high temperature tolerance of C. glycerinogenes. Expression of CgMCUR1 resulted in better acetic acid, H2O2, and high temperature tolerance in recombinant S. cerevisiae. Meanwhile, CgMCUR1 was able to enhance intracellular proline accumulation. The qRT-PCR analysis revealed that overexpression of CgMCUR1 affected proline metabolism in recombinant S. cerevisiae. The overexpression strain also showed reduced levels of cellular lipid peroxidation and an altered ratio of saturated fatty acid (SFA) to unsaturated fatty acid (UFA) in the cell membrane. The ethanol production of recombinant S. cerevisiae at high temperature was 30.9 g l-1, obtaining an increase of 12%, and the conversion rate was increased by 12%. In the undetoxified cellulose hydrolysate, the ethanol yield was 14.7 g l-1 at 30 h with an improvement of 18.5%, and the conversion rate was increased by 15.3%.
CONCLUSIONS
Overexpression of CgMCUR1 rendered the acetic acid, H2O2, and high temperature tolerant of recombinant S. cerevisiae and enhanced the ethanol fermentation performance of recombinant S. cerevisiae under high temperature stress and in undetoxified cellulose hydrolysate by improving intracellular proline accumulation and by changing cellular physiological metabolism.
Topics: Saccharomyces cerevisiae; Hydrogen Peroxide; Saccharomyces cerevisiae Proteins; Ethanol; Fermentation; Cellulose; Acetic Acid; Proline
PubMed: 37307223
DOI: 10.1093/jambio/lxad108 -
ACS Synthetic Biology Jun 2023Geraniol is a class of natural products that are widely used in the aroma industry due to their unique aroma. Here, to achieve the synthesis of geraniol and alleviate...
Geraniol is a class of natural products that are widely used in the aroma industry due to their unique aroma. Here, to achieve the synthesis of geraniol and alleviate the intense competition from the yeast ergosterol pathway, a transcription factor-mediated ergosterol feedback system was developed in this study to autonomously regulate ergosterol metabolism and redirect carbon flux to geraniol synthesis. In addition, the modification of ergosterol-responsive promoters, the optimization of transcription factor expression intensity, and stepwise metabolic engineering resulted in a geraniol titer of 531.7 mg L. For sustainable production of geraniol, we constructed a xylose assimilation pathway in (). Then, the xylose metabolic capacity was ameliorated and the growth of the engineered strain was rescued by activating the pentose phosphate (PP) pathway. Finally, we obtained 1091.6, 862.4, and 921.8 mg L of geraniol in a 5 L bioreactor by using pure glucose, simulated wheat straw hydrolysates, and simulated sugarcane bagasse hydrolysates, with yields of 47.5, 57.9, and 59.1 mg g DCW, respectively. Our study demonstrated that has the potential to produce geraniol from lignocellulosic biomass, providing a powerful tool for the sustainable synthesis of other valuable monoterpenes.
Topics: Cellulose; Metabolic Engineering; Xylose; Fermentation; Saccharum; Transcription Factors
PubMed: 37271978
DOI: 10.1021/acssynbio.3c00195 -
Journal of Water and Health May 2023Cockroaches are known as mechanical vectors of some pathogens that can infect humans. The present study aims to rapidly identify Periplaneta americana fungal pathogens...
Cockroaches are known as mechanical vectors of some pathogens that can infect humans. The present study aims to rapidly identify Periplaneta americana fungal pathogens from sewer systems of public hospitals in Esfahan using the polymerase chain reaction (PCR) technique. A total of 55 P. americana cockroaches were randomly collected by direct trapping from sewer systems of seven hospitals and screened for fungal infectious agents using standard morphological methods and the PCR sequencing. From the American cockroach, we isolated 62 yeasts and 31 molds from the surface, hemocoel, and digestive tract of P. americana. Based on DNA sequence comparisons and other taxonomic characteristics, they were identified as more than four species of yeast and four species of mold. Yeast species including Pichia kudriavzevii, Candida glabrata, Pichia kluyveri, and Candida viswanathii, and molds such as Aspergillus niger, Penicillium italicum, Mucor plumbeus, and Rhizopus oryzae were isolated repeatedly from the surface, hemocoel, and digestive tract of P. americana. Our results show that the use of a combination of morphological, molecular techniques, and phylogenetic analysis can lead to the identification of pathogenic fungal agents in American cockroaches and also knowledge of fungal pathogens-arthropod host relationships.
Topics: Animals; Humans; Cockroaches; Periplaneta; Phylogeny; Fungi; Hospitals
PubMed: 37254911
DOI: 10.2166/wh.2023.015 -
Journal of Agricultural and Food... Jun 2023Caffeic acid is a phenolic acid compound widely applied in the food and pharmaceutical fields. Currently, one of the reasons for the low yield of caffeic acid...
Caffeic acid is a phenolic acid compound widely applied in the food and pharmaceutical fields. Currently, one of the reasons for the low yield of caffeic acid biosynthesis is that the carbon flow enters mainly into the TCA cycle via pyruvate, which leads to low concentrations of erythrose 4-phosphate (E4P) and phosphoenolpyruvate (PEP), the precursors of caffeic acid synthesis. Here, we developed a growth-coupled dual-layered dynamic regulation system. This system controls intracellular pyruvate supply in real time by responding to intracellular pyruvate and -coumaric acid concentrations, autonomously coordinates pathway gene expression, and redirects carbon metabolism to balance cell growth and caffeic acid synthesis. Finally, our constructed engineered strain based on the dual-layered dynamic regulation system achieved a caffeic acid titer of 559.7 mg/L in a 5 L bioreactor. Thus, this study demonstrated the efficiency and potential of this system in boosting the yield of aromatic compounds.
Topics: Pyruvic Acid; Caffeic Acids; Carbon
PubMed: 37254503
DOI: 10.1021/acs.jafc.3c01722 -
Food Research International (Ottawa,... Jul 2023Ethyl hexanoate and ethyl butyrate are essential to the flavor compounds in Nongxiangxing baijiu, but low levels of these two esters in upper fermented grains (FG)...
Application of Clostridium butyricum, Rummeliibacillus suwonensis, and Issatchenkia orientalis for Nongxiangxing baijiu fermentation: Improves the microbial communities and flavor of upper fermented grain.
Ethyl hexanoate and ethyl butyrate are essential to the flavor compounds in Nongxiangxing baijiu, but low levels of these two esters in upper fermented grains (FG) decreases the quality of upper distilled baijiu, representing the main challenge in Nongxiangxing baijiu production. This paper enhanced fermentation by inoculating functional Clostridium butyricum, Rummeliibacillus suwonensis, and Issatchenkia orientalis strains into upper FG. The results showed that the ethyl butyrate content in the upper FG increased significantly and the content of ethyl hexanoate did improve from the results of many determinations. High-throughput sequencing indicated that the dominant phyla in the FG were Firmicutes, Actinobacteriota, Proteobacteria, Ascomycota, and Basidiomycota. The canonical correspondence analysis (CCA) and person correlation network revealed the relationship between the microbial community, physicochemical environment, and flavor compounds. The temperature, oxygen, and acidity were closely related to the microbial community, while most flavor compounds were positively correlated with Caldicoprobacter, Caproiciproducens, Delftia, Hydrogenispora, Thermoactinomyces, Issatchenkia Bacillus, and Aspergillus. These results helped improve the quality of Nongxiangxing baijiu.
Topics: Humans; Alcoholic Beverages; Clostridium butyricum; Fermentation; Microbiota
PubMed: 37254333
DOI: 10.1016/j.foodres.2023.112885 -
International Journal of Food... Aug 2023The increasing demand for more flavored and complex beers encourages the investigation of novel and non-conventional yeasts with the ability to provide a combination of...
The increasing demand for more flavored and complex beers encourages the investigation of novel and non-conventional yeasts with the ability to provide a combination of bioflavoring and low ethanol yields. The present study identified 22 yeasts isolated from different brewing sources, including the fermentation by-products known as yeast sludges, and characterized a selection of strains to find the more suitable for the aforementioned aims. HPLC and GC-FID analysis of its brewing products were performed. The most promising results were obtained with the non-conventional yeasts Pichia kudriavzevii MBELGA61 and Meyerozyma guilliermondii MUS122. The former, isolated from a Belgian wheat beer sludge, was capable of growing in wort (17.0°Bx., 20 °C) with very low ethanol yields (1.19 % v/v). Besides, upon mixed fermentations with Saccharomyces cerevisiae, was suitable to produce volatile compounds such as ethyl acetate, 2-phenyl ethanol and isoamyl alcohol, with characteristic fruity notes. M. guilliermondii MUS122, isolated from a golden ale beer sludge, partially attenuated the wort with low production of ethanol and biomass. In addition, provided some fruity and floral nuances to the aroma profile of mixed fermentations with brewer's yeast. The results suggest that these strains favor the development of more fruity-flowery aroma profiles in beers. Furthermore, they are suitable for use in mixed fermentations with Saccharomyces brewer's strains, although the ethanol level did not decrease significantly.
Topics: Fermentation; Beer; Sewage; Yeasts; Saccharomyces cerevisiae; Ethanol
PubMed: 37244227
DOI: 10.1016/j.ijfoodmicro.2023.110254 -
Foods (Basel, Switzerland) May 2023Cocoa bean fermentation is carried out in different production areas following various methods. This study aimed to assess how the bacterial and fungal communities were...
Cocoa bean fermentation is carried out in different production areas following various methods. This study aimed to assess how the bacterial and fungal communities were affected by box, ground or jute fermentation methods, using high-throughput sequencing (HTS) of phylogenetic amplicons. Moreover, an evaluation of the preferable fermentation method was carried out based on the microbial dynamics observed. Box fermentation resulted in higher bacterial species diversity, while beans processed on the ground had a wider fungal community. and were observed in all three fermentation methods studied. Moreover, dominated box fermentation and abounded in ground-fermented samples. was the most important yeast in jute and box, while prevailed in the box and ground fermentation. PICRUST analysis was performed to identify potential interesting pathways. In conclusion, there were noticeable differences between the three different fermentation methods. Due to its limited microbial diversity and the presence of microorganisms that guarantee good fermentation, the box method was found to be preferable. Moreover, the present study allowed us to thoroughly explore the microbiota of differently treated cocoa beans and to better understand the technological processes useful to obtain a standardized end-product.
PubMed: 37238842
DOI: 10.3390/foods12102024