Alicia Maciá Valero, Jeroen J van Wageningen, Alexander J Foster, Ana Rita Oliveira, Clemens Mayer, Sonja Billerbeck
{"title":"Pulcherriminic acid Biosynthesis and Transport: Insights from a heterologous system in Saccharomyces cerevisiae.","authors":"Alicia Maciá Valero, Jeroen J van Wageningen, Alexander J Foster, Ana Rita Oliveira, Clemens Mayer, Sonja Billerbeck","doi":"10.1093/femsyr/foaf039","DOIUrl":"https://doi.org/10.1093/femsyr/foaf039","url":null,"abstract":"<p><p>Pulcherriminic acid is an iron chelator produced by some Kluyveromyces and Metschnikowia yeasts. Its biosynthesis is encoded by the four-gene PUL cluster, where PUL1 and PUL2 are the biosynthetic enzymes, PUL3 mediates the uptake of iron-bound pulcherrimin, and PUL4 is a putative regulator. Pulcherriminic acid holds antifungal potential, as the growth of organisms unable to uptake pulcherrimin is inhibited by deficit of essential iron. Thus, a heterologous production system to further characterize and optimize its biosynthesis would be valuable. Using our in-house yeast collection and genomes available in databases, we cloned PUL1 and PUL2 genes from K. lactis and one of our wild Metschnikowia isolates and built an effective production system in S. cerevisiae able to inhibit pathogenic growth. In this context, the K. lactis genes yielded faster pulcherriminic acid production than those from the Metschnikowia isolate and a combinatorial approach showed PUL1 to be the production bottleneck. We further showed that Pul3 is an importer of pulcherrimin, but also mediates the export of pulcherriminic acid and that the growth of pathogens like Candidozyma auris and organisms encoding PUL3 in their genome, previously called \"cheaters\", is inhibited by pulcherriminic acid, highlighting its potential as an antimicrobial agent.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144729069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Glycerol Bioconversion to Biofuel and Value-Added Products by Yeasts.","authors":"Kostyantyn Dmytruk, Marta Semkiv, Andriy Sibirny","doi":"10.1093/femsyr/foaf038","DOIUrl":"https://doi.org/10.1093/femsyr/foaf038","url":null,"abstract":"<p><p>Glycerol, a by-product of biodiesel production, is a versatile polyol used in various industries. Yeasts play a crucial role in converting glycerol into biofuels and value-added products, offering sustainable alternatives to chemical synthesis. This review explores glycerol metabolism in yeasts, focusing on its bioconversion into ethanol, isopropanol, lipids, organic acids, and polyols. Saccharomyces cerevisiae and Yarrowia lipolytica are prominent species for these processes, with metabolic engineering enhancing their efficiency. Ethanol production from crude glycerol, a by-product of the biodiesel industry, is cost-effective compared to traditional feedstocks, while lipid production by oleaginous yeasts supports biodiesel synthesis. Organic acids like succinic, citric, and lactic acids, along with polyols such as erythritol and mannitol, are produced through optimized pathways, achieving high yields. Crude glycerol, despite impurities, is a viable low-cost substrate, with yeast strains adapted to tolerate its contaminants. Challenges include improving strain tolerance and scaling up processes. Future research aims to refine metabolic engineering and fermentation strategies to maximize glycerol's potential as a renewable feedstock for industrial biotechnology.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144689695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hadeel A B Elnaim Mohamed, Hizlan Hincal Agus, Bedia Palabiyik
{"title":"A novel method for telomere length detection in fission yeast.","authors":"Hadeel A B Elnaim Mohamed, Hizlan Hincal Agus, Bedia Palabiyik","doi":"10.1093/femsyr/foae040","DOIUrl":"10.1093/femsyr/foae040","url":null,"abstract":"<p><p>Fission yeast is the ideal model organism for studying telomere maintenance in higher eukaryotes. Telomere length has been directly correlated with life expectancy and the onset of aging-related diseases in mammals. In this study, we developed a novel simple, and reproducible method to measure the telomere length, by investigating the effect of caffeine and cisplatin on the telomere length in fission yeast. Hydroxyurea-synchronized fission yeast cells were exposed to 62 µM cisplatin and 8.67 mM caffeine treatments for 2 h, then their telomere lengths were evaluated with two different methods. First, the quantitative polymerase chain reaction (qPCR) assay was used as a confirmative method, where telomere length was determined relative to a single-copy gene in the genome. Second, the newly developed method standard polymerase chain reaction (PCR)/ImageJ assay assessed the telomere length based on the amplified PCR band intensity using a set of telomere primers, reflecting telomeric sequence availability in the genome. Both methods show a significant decrease and a notable telomere lengthening in response to cisplatin and caffeine treatments, respectively. The finding supports the accuracy and productivity of the standard PCR/ImageJ assay as it can serve as a quick screening tool to study the effect of suspected chemotherapeutic and antiaging drugs on telomere length in fission yeast.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11781191/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142885275","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The advances in creating Crabtree-negative Saccharomyces cerevisiae and the application for chemicals biosynthesis.","authors":"Yalin Guo, Zhen Xiong, Haotian Zhai, Yuqi Wang, Qingsheng Qi, Jin Hou","doi":"10.1093/femsyr/foaf014","DOIUrl":"10.1093/femsyr/foaf014","url":null,"abstract":"<p><p>Saccharomyces cerevisiae is a promising microbial cell factory. However, the overflow metabolism, known as the Crabtree effect, directs the majority of the carbon source toward ethanol production, in many cases, resulting in low yields of other target chemicals and byproducts accumulation. To construct Crabtree-negative S. cerevisiae, the deletion of pyruvate decarboxylases and/or ethanol dehydrogenases is required. However, these modifications compromises the growth of the strains on glucose. This review discusses the metabolic engineering approaches used to eliminate ethanol production, the efforts to alleviate growth defect of Crabtree-negative strains, and the underlying mechanisms of the growth rescue. In addition, it summarizes the applications of Crabtree-negative S. cerevisiae in the synthesis of various chemicals such as lactic acid, 2,3-butanediol, malic acid, succinic acid, isobutanol, and others.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11974387/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143691625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cristobal A Onetto, Chris M Ward, Cristian Varela, Laura Hale, Simon A Schmidt, Anthony R Borneman
{"title":"Genetic and phenotypic diversity of wine-associated Hanseniaspora species.","authors":"Cristobal A Onetto, Chris M Ward, Cristian Varela, Laura Hale, Simon A Schmidt, Anthony R Borneman","doi":"10.1093/femsyr/foaf031","DOIUrl":"10.1093/femsyr/foaf031","url":null,"abstract":"<p><p>The genus Hanseniaspora includes apiculate yeasts commonly found in fruit- and fermentation-associated environments. Their genetic diversity and evolutionary adaptations remain largely unexplored despite their ecological and oenological significance. This study investigated the phylogenetic relationships, genome structure, selection patterns, and phenotypic diversity of Hanseniaspora species isolated primarily from Australian wine environments, focusing on Hanseniaspora uvarum, the most abundant non-Saccharomyces yeast in wine fermentation. A total of 151 isolates were sequenced, including long-read genomes for representatives of the main phylogenetic clades. Comparative genomics revealed ancestral chromosomal rearrangements between the slow-evolving lineage (SEL) and fast-evolving lineage (FEL) that could have contributed to their evolutionary split, as well as significant loss of genes associated with mRNA splicing, chromatid segregation and signal recognition particle protein targeting in the FEL. Pangenome analysis within H. uvarum identified extensive copy number variation, particularly in genes related to xenobiotic tolerance and nutrient transport. Investigation into the selective landscape following the FEL/SEL divergence identified diversifying selection in 229 genes in the FEL, with significant enrichment in genes within the lysine biosynthetic pathway. Furthermore, phenotypic screening of 116 isolates revealed substantial intraspecific diversity, with specific species exhibiting enhanced ethanol, osmotic, copper, SO₂, and cold tolerance.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12199727/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144265815","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Recent advances in genetic engineering and chemical production in yeast species.","authors":"Sangdo Yook, Hal S Alper","doi":"10.1093/femsyr/foaf009","DOIUrl":"10.1093/femsyr/foaf009","url":null,"abstract":"<p><p>Yeasts have emerged as well-suited microbial cell factory for the sustainable production of biofuels, organic acids, terpenoids, and specialty chemicals. This ability is bolstered by advances in genetic engineering tools, including CRISPR-Cas systems and modular cloning in both conventional (Saccharomyces cerevisiae) and non-conventional (Yarrowia lipolytica, Rhodotorula toruloides, Candida krusei) yeasts. Additionally, genome-scale metabolic models and machine learning approaches have accelerated efforts to create a broad range of compounds that help reduce dependency on fossil fuels, mitigate climate change, and offer sustainable alternatives to petrochemical-derived counterparts. In this review, we highlight the cutting-edge genetic tools driving yeast metabolic engineering and then explore the diverse applications of yeast-based platforms for producing value-added products. Collectively, this review underscores the pivotal role of yeast biotechnology in efforts to build a sustainable bioeconomy.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11963765/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143623951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Integrated omic analysis of a new flavor yeast strain in fermented rice milk.","authors":"Chayaphathra Sooklim, Atchara Paemanee, Khanok Ratanakhanokchai, Duanghathai Wiwatratana, Nitnipa Soontorngun","doi":"10.1093/femsyr/foaf017","DOIUrl":"10.1093/femsyr/foaf017","url":null,"abstract":"<p><p>Plant-based milk contains high nutritional value with enriched vitamins, minerals, and essential amino acids. This study aimed to enhance the biochemical and biological properties of rice milk through yeast fermentation, using the novel fermenting strain Saccharomyces cerevisiae RSO4, which has superb fermenting ability for an innovative functional beverage. An integrated omics approach identified specific genes that exhibited genetic variants related to various cellular processes, including flavor and aroma production (ARO10, ADH1-5, and SFA1), whereas the proteomic profiles of RSO4 identified key enzymes whose expression was upregulated during fermentation of cooked rice, including the enzymes in glycogen branching (Glc3), glycolysis (Eno1, Pgk1, and Tdh1/2), stress response (Hsp26 and Hsp70), amino acid metabolism, and cell wall integrity. Biochemical and metabolomic analyses of the fermented rice milk by the RSO4 strain using the two rice varieties, Homali (Jasmine) white rice or Riceberry colored rice, detected differentially increased levels of bioactive compounds, such as β-glucan, vitamins, di- and tripeptides, as well as pleasant flavors and aromas. The results of this study highlight the importance of selecting an appropriate fermenting yeast strain and rice variety to improve property of plant-based products as innovative functional foods.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11995695/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143735674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"BAHD acyltransferase from dragon fruit enables production of phyllocactin in engineered yeast.","authors":"Christiane Glitz, Jane Dannow Dyekjær, Sophia Mattitsch, Mahsa Babaei, Irina Borodina","doi":"10.1093/femsyr/foae041","DOIUrl":"10.1093/femsyr/foae041","url":null,"abstract":"<p><p>Microbial fermentation can provide a sustainable and cost-effective alternative to traditional plant extraction to produce natural food colours. Betalains are a class of yellow to red water-soluble pigments. Even though over 80 betalain variants are known, betanin is the only betalain available as a food colourant on the market. Many variants are acylated, which can enhance their stability and change the hue, but very few acyltransferases responsible for the acylation are known. Therefore, we mined the transcriptomes of Celosia argentea var. cristata and Hylocereus polyrhizus for BAHD acyltransferases, enzymes likely involved in betalain acylation. In vivo screening of the enzymes in betanin-producing Saccharomyces cerevisiae revealed that the acyltransferase HpBAHD3 from H. polyrhizus malonylates betanin, forming phyllocactin (6'-O-malonyl-betanin). This is the first identification of a BAHD acyltransferase involved in betalain biosynthesis. Expression of HpBAHD3 in a Yarrowia lipolytica strain engineered for high betanin production led to near-complete conversion of betanin to phyllocactin. In fed-batch fermentation, the strain produced 1.95 ± 0.024 g/l phyllocactin in 60 h. This study expands the range of natural food colourants produced through microbial fermentation and contributes to elucidating the biosynthesis pathway of acylated betalains.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11881927/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143390432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Franco Vega-Macaya, Pablo Villarreal, Tomas A Peña, Valentina Abarca, Agustín A Cofré, Christian I Oporto, Wladimir Mardones, Roberto F Nespolo, Francisco A Cubillos
{"title":"Experimental evolution and hybridization enhance the fermentative capacity of wild Saccharomyces eubayanus strains.","authors":"Franco Vega-Macaya, Pablo Villarreal, Tomas A Peña, Valentina Abarca, Agustín A Cofré, Christian I Oporto, Wladimir Mardones, Roberto F Nespolo, Francisco A Cubillos","doi":"10.1093/femsyr/foaf004","DOIUrl":"10.1093/femsyr/foaf004","url":null,"abstract":"<p><p>Lager beer is traditionally fermented using Saccharomyces pastorianus. However, the limited availability of lager yeast strains restricts the potential range of beer profiles. Recently, Saccharomyces eubayanus strains showed the potential to impart novel aromas to beer, with slower fermentation rates than commercial strains. Here, we applied experimental evolution to nine S. eubayanus strains using three different selective conditions to generate improved strains to fermentative environments. We observed environment-dependent fitness changes across strains, with ethanol-enriched media resulting in the greatest fitness improvement. We identified subtelomeric genomic changes in a deficient fermentative strain underlying the greatest fitness improvement. Gene expression analysis and genome sequencing identified genes associated with oxidative stress, amino acid metabolism, sterol biosynthesis, and vacuole morphology underlying differences between evolved and the ancestral strain, revealing the cellular processes underlying fermentation improvement. A hybridization strategy between two evolved strains allowed us to expand the phenotypic space of the F2 segregants, obtaining strains with a 13.7% greater fermentative capacity relative to the best evolved parental strains. Our study highlights the potential of integrating experimental evolution and hybridization to enhance the fermentation capacity of wild yeast strains, offering strengthened solutions for industrial applications and highlighting the potential of Patagonian S. eubayanus in brewing.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11878536/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143064865","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Molecular evolution of the members of the Snq2/Pdr18 subfamily of Pdr transporters in the Hemiascomycete yeasts.","authors":"Paulo Jorge Dias","doi":"10.1093/femsyr/foaf026","DOIUrl":"10.1093/femsyr/foaf026","url":null,"abstract":"<p><p>The transporters of the ATP-Binding Cassette (ABC) Superfamily involved in the Multidrug Resistance (MDR) phenomena are also known as ABC-Pleiotropic Drug Resistance (PDR) proteins. The homologs of the Saccharomyces cerevisiae SNQ2 and PDR18 genes were identified in 171 yeast genomes, representing 68 different hemiascomycetous species. All early-divergent yeast species analyzed in this work lack Snq2/Pdr18 homologs, suggesting that the origin of these ABC-PDR genes in hemiascomycete yeasts resulted from a horizontal transfer event. The evolutionary pathway of the Snq2/Pdr18 protein subfamily in pathogenic Candida species was also reconstructed, revealing a main gene lineage leading to the Candida albicans SNQ2 gene. The results indicate that, after the gene duplication event at the origin of the SNQ2/PDR18 paralogs, the PDR18 ortholog has been under strong diversifying selection and suggest that a small portion of the sequence of the SNQ2 ancestral ortholog might have been under mild positive selection. The results also showed that strong positive selection was exerted over one of the two paralogs generated by the Whole Genome Duplication (WGD) event, corresponding to the duplicate at the origin of a \"short-lived\" WGD sublineage.</p>","PeriodicalId":12290,"journal":{"name":"FEMS yeast research","volume":" ","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12202755/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144157638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}