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FEMS Yeast Research

Paulo César Silva, Lucília Domingues, Tony Collins, Rui Oliveira, Björn Johansson
Lignocellulosic hydrolysates remain one of the most abundantly used substrates for the sustainable production of second generation fuels and chemicals with Saccharomyces cerevisiae. Nevertheless, fermentation inhibitors such as acetic acid, furfural and hydroxymethylfurfural are formed during the process and can lead to slow or stuck fermentations and/or act as genotoxic agents leading to production strain genetic instability. We have developed a novel dominant deletion (DEL) cassette assay for quantification of DNA damage in both wild-type and industrial yeast strains...
September 14, 2018: FEMS Yeast Research
Miguel Fernández-Niño, Sergio Pulido, Despina Stefanoska, Camilo Pérez, Daniel González-Ramos, Antonius J A van Maris, Kathleen Marchal, Elke Nevoigt, Steve Swinnen
Acetic acid tolerance of the yeast Saccharomyces cerevisiae is manifested in several quantifiable parameters, of which the duration of the latency phase is one of the most studied. It has been shown recently that the latter parameter is mostly determined by a fraction of cells within the population that resumes proliferation upon exposure to acetic acid. The aim of the current study was to identify genetic determinants of the difference in this parameter between the highly tolerant strain MUCL 11987-9 and the laboratory strain CEN...
September 14, 2018: FEMS Yeast Research
Jiajing Wang, Haotian Zhai, Reheman Rexida, Yu Shen, Jin Hou, Xiaoming Bao
Fine-tuning of the expression of genes are crucial for cell factory construction. Promoters are the most important tools to control gene expression. However, native promoters are often limited by their transcriptional ability. In this study, we sought to overcome the limitations of native promoters in Saccharomyces cerevisiae through the construction of hybrid promoter libraries for both constitutive promoters and promoters induced by diauxic shift. A series of hybrid constitutive promoters were constructed by combing the upstream activation sequences (UAS) and changing the core promoter elements...
September 10, 2018: FEMS Yeast Research
Susana Perrusquía-Luévano, Marlene Sarahí Cano-Herrera, César Guigón-López, María Del Carmen Avitia-Talamantes, Carlos Torres-Torres, Isaela Villalpando
This work reports important results of aromatic profiles produced by native yeasts isolated from desert-grown wine grapes from the south of Chihuahua, México. These grapes stand very high temperatures during the ripening season, developing high sugar concentration and high pH. Yeast species found in grapes were identified by polymerase chain reaction and sequence analysis of the 5.8S internal transcribed spacer ribosomal RNA region. Aureobasidium namibiae, Sporobolomyces johnsonii, Candida apicola, Hanseniaspora uvarum, Candida thaimueangensis, Hanseniaspora opuntiae, were identified...
September 10, 2018: FEMS Yeast Research
Joana Tulha, Cândida Lucas
GUP1 is the yeast counterpart of the high eukaryotes HHATL. This and the close homologue Gup2/HHAT regulate the Hedgehog morphogenic, developmental pathway. In yeasts, a similar paracrine pathway is not known though the Δgup1 mutant is associated with morphology and proliferation/death processes. As a first step towards identifying the actual molecular/enzymatic function of Gup1, this work identified by co-immunoprecipitation the yeast mitochondria membrane VDAC1/Por1 as a physical partner of Gup1. Gup1 locates in the ER and plasma membrane...
September 3, 2018: FEMS Yeast Research
Michael Gossing, Agata Smialowska, Jens Nielsen
Nutrient sensing and signaling controls the cellular response to extracellular nutrients and intracellular metabolites. Nutrient-dependent regulation of metabolism ensures balanced energy production and expenditure. We show that disturbing energy balance by forcing fatty acid synthesis has profound impact on metabolism and physiology of the yeast cell. In addition to an expected increase in storage lipids, we observed increased β-oxidation and reduced amino acid biosynthesis, indicating increased activity of nutrient-sensitive kinase Snf1p...
August 29, 2018: FEMS Yeast Research
Xinchen Teng, Eric Yau, Cierra Sing, J Marie Hardwick
Cells are exquisitely tuned to environmental ques. Amino acid availability is rapidly sensed, allowing cells to adjust molecular processes and implement short or long term metabolic shifts accordingly. How levels of most individual amino acids may be sensed and subsequently signaled to inform cells of their nutrient status is largely unknown. We made the unexpected observation that small changes in the levels of specific amino acids can have a profound effect on yeast cell growth, leading to the identification of yeast Whi2 as a negative regulator of cell growth in low amino acids...
August 27, 2018: FEMS Yeast Research
Pallavi Kakade, Kasturi Mahadik, Kithiganahalli Narayanaswamy Balaji, Kaustuv Sanyal, Valakunja Nagaraja
Candida albicans, a human pathogen, carries an expanded family of Zn(II)2Cys6 transcription factors. A CTG clade-specific protein Zcf32 and its closely related protein Upc2, a well-conserved transcription factor across the various fungal species, belong to this family of proteins. Unlike Upc2, Zcf32 is poorly studied in C. albicans. Here, we examined roles played by these two related transcription factors in biofilm development and virulence of C. albicans. Our data show that the null mutants of each of Zcf32 or Upc2 form better biofilms than the wild-type suggesting that both of them negatively regulate the biofilm development...
July 24, 2018: FEMS Yeast Research
Maarten D Verhoeven, Sophie C de Valk, Jean-Marc G Daran, Antonius J A van Maris, Jack T Pronk
d-Glucose, d-xylose and l-arabinose are major sugars in lignocellulosic hydrolysates. This study explores fermentation of glucose-xylose-arabinose mixtures by a consortium of three 'specialist' Saccharomyces cerevisiae strains. A d-glucose- and l-arabinose-tolerant xylose specialist was constructed by eliminating hexose phosphorylation in an engineered xylose-fermenting strain and subsequent laboratory evolution. A resulting strain anaerobically grew and fermented d-xylose in the presence of 20 g L-1 of d-glucose and l-arabinose...
July 16, 2018: FEMS Yeast Research
Sergio Giannattasio, Cristina Mazzoni
No abstract text is available yet for this article.
December 1, 2018: FEMS Yeast Research
Monika Cioch-Skoneczny, Pawel Satora, Magdalena Skotniczny, Szymon Skoneczny
The aim of the study was to characterise, quantitatively and qualitatively, the yeast microbiota found during spontaneous fermentation of grape musts obtained from the red grape varieties 'Rondo' and 'Regent'. For the research, grapes originating from two vineyards (Srebrna Góra and Spotkaniówka) located in southern Poland were used. Musts of the studied grape varieties provided a favourable environment for the growth of microorganisms. Musts obtained from fruits from the Spotkaniówka vineyard were characterised by a higher overall number of yeasts...
December 1, 2018: FEMS Yeast Research
Nicoletta Guaragnella, Liam P Coyne, Xin Jie Chen, Sergio Giannattasio
Mitochondria are key cell organelles with a prominent role in both energetic metabolism and the maintenance of cellular homeostasis. Since mitochondria harbor their own genome, which encodes a limited number of proteins critical for oxidative phosphorylation and protein translation, their function and biogenesis strictly depend upon nuclear control. The yeast Saccharomyces cerevisiae has been a unique model for understanding mitochondrial DNA organization and inheritance as well as for deciphering the process of assembly of mitochondrial components...
December 1, 2018: FEMS Yeast Research
Laura K F McNair, Solvej Siedler, Joachim M O Vinther, Anna Mette Hansen, Ana Rute Neves, Christel Garrigues, Anna K Jäger, Henrik Franzyk, Dan Staerk
Mold and yeast contamination constitutes a major problem in food commodities, including dairy products, hence new natural preventive measures are in high demand. The aim of the current study is to identify and characterize novel antifungal peptides produced by lactic acid bacteria (LAB) in sour cream. By the use of a newly developed image-based 96-well plate fungal growth inhibition assay targeting Debaryomyces hansenii, combined with a range of analytical tools comprising HPLC-high-resolution mass spectrometry, ultrahigh-performance liquid chromatography-Triple Quadrupole MS and nuclear magnetic resonance spectroscopy, we successfully identified a new antifungal peptide (DMPIQAFLLY; 1211 Da) in sour cream enriched with two bioprotective LAB strains...
December 1, 2018: FEMS Yeast Research
Jessica A Zinskie, Meghan Roig, Christopher Janetopoulos, Kenneth A Myers, Michael F Bruist
The development of the RNA 'vegetable' aptamers, Spinach and Broccoli, has simplified RNA imaging, especially in live cells. These RNA aptamers interact with a fluorophore (DFHBI or DFHBI-1T) to produce a green fluorescence signal. Although used in mammalian and Escherichia coli cells, the use of these aptamers in yeast has been limited. Here we describe how the Saccharomyces cerevisiae snoRNA, snR30, was tagged with the Spinach or the Broccoli aptamers and observed in live cells. The ability to observe aptamer fluorescence in polyacrylamide gels stained with a fluorophore or with a microplate reader can ease preliminary screening of the aptamers in different RNA scaffolds...
December 1, 2018: FEMS Yeast Research
Barbara Knoblach, Richard A Rachubinski
We report the permanent introduction of the human peroxisomal β-oxidation enzymatic machinery required for straight chain degradation of fatty acids into the yeast, Saccharomyces cerevisiae. Peroxisomal β-oxidation encompasses four sequential reactions that are confined to three enzymes. The genes encoding human acyl-CoA oxidase 1, peroxisomal multifunctional enzyme type 2 and 3-ketoacyl-CoA thiolase were introduced into the genomic loci of their yeast gene equivalents. The human β-oxidation genes were individually tagged with sequence coding for GFP and expression of the protein chimeras as well as their targeting to peroxisomes was confirmed...
December 1, 2018: FEMS Yeast Research
Xiaofei Song, Yuanzi Li, Yuzhen Wu, Miao Cai, Quanli Liu, Kai Gao, Xiuming Zhang, Yanling Bai, Haijin Xu, Mingqiang Qiao
As a traditional ethanol-producing microorganism, Saccharomyces cerevisiae is an ideal host for consolidated bioprocessing. However, expression of heterologous cellulase increases the metabolic burden in yeast, which results in low cellulase activity and poor cellulose degradation efficiency. In this study, cellulase-expressing yeast strains that could efficiently degrade different cellulosic substrates were created by optimizing cellulase ratios through a POT1-mediated δ-integration strategy. Metabolic engineering strategies, including optimization of codon usage, promoter and signal peptide, were also included in this system...
December 1, 2018: FEMS Yeast Research
Liming Ouyang, Petter Holland, Hongzhong Lu, David Bergenholm, Jens Nielsen
The Saccharomyces cerevisiae transcription factor (TF) Stb5 is known to be involved in regulating NADPH generation. We explored its role by combining DNA binding studies with transcriptome analysis at four environmental conditions that were selected to cover a range of different metabolic states. Using ChIP-exo, DNA binding targets of Stb5 were found to confirm many previously proposed binding targets, in particular genes encoding enzymes involved in NADPH generation and the pentose-phosphate (PP) pathway. Transcriptome analysis of an STB5 deletion strain revealed transcriptional changes in direct regulation targets of Stb5, including several PP pathway genes as well as additional novel regulatory targets, but interestingly not including the proposed PP pathway flux controlling enzyme Zwf1...
December 1, 2018: FEMS Yeast Research
Larissa V G Longo, Stephanie C Ray, Rosana Puccia, Chad A Rappleye
The fungal APSES protein family of transcription factors is characterized by a conserved DNA-binding motif facilitating regulation of gene expression in fungal development and other biological processes. However, their functions in the thermally dimorphic fungal pathogen Histoplasma capsulatum are unexplored. Histoplasma capsulatum switches between avirulent hyphae in the environment and virulent yeasts in mammalian hosts. We identified five APSES domain-containing proteins in H. capsulatum homologous to Swi6, Mbp1, Stu1 and Xbp1 proteins and one protein found in related Ascomycetes (APSES-family protein 1; Afp1)...
December 1, 2018: FEMS Yeast Research
Xiang Jiao, Qi Zhang, Sufang Zhang, Xiaobing Yang, Qian Wang, Zongbao Kent Zhao
The red yeast Rhodosporidium toruloide is a versatile host for production of lipids and carotenoids. Genetic tools are underdeveloped for red yeasts due to their unique genetics and physiology. Currently expression of a heterogonous gene in red yeasts is largely based on integration of the designed cassette by Agrobacterium mediated transformation, yet this method is somewhat restricted when multiple genes are required to be expressed due to the lack of functional genetic elements. Here we demonstrate that virus 2A sequence is effective to mediate co-expression of multiple enzymes in R...
December 1, 2018: FEMS Yeast Research
Birgitta E Ebert, Eik Czarnotta, Lars M Blank
Heterologous synthesis of triterpenoids in Saccharomyces cerevisiae from its native metabolite squalene has been reported to offer an alternative to chemical synthesis and extraction from plant material if productivities can be increased.Here, we physiologically characterized a squalene overproducing S. cerevisiae CEN.PK strain to elucidate the effect of cultivation conditions on the production of this central triterpenoid precursor. The maximum achievable squalene concentration was substantially influenced by nutritional conditions, medium composition and cultivation mode...
December 1, 2018: FEMS Yeast Research
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