Comparative Activity of Six Recombinant Stilbene Synthases in Yeast for Resveratrol Production

Resveratrol is a nutraceutical with relevant benefits to human health. This investigation reports on the generation and evaluation of six recombinant yeast lines that produce resveratrol from p-coumaric acid. The yeast lines contained a single p-coumaric acid-Co-A ligase from Plagiochasma appendiculatum combined with the stilbene synthases from Parthenocissus henryana, Polygonum cuspidatum, Morus alba var. atropurpurea, Rheum tataricum, Vitis vinifera and Arachis hypogaea. Codon optimized versions of these sequences were inserted in an expression vector flanked by the constitutive PGK and GPD promoters before expression in Saccharomyces cerevisiae. Batch fermentation (60 h) revealed that yeast lines had different capacities (p < 0.01) to produce resveratrol. Slightly acidic pH (6) and concentrations <100 mg L−1 p-coumaric acid improved resveratrol yields. Among the six lines, those containing the stilbene synthases (STS) from P. cuspidatum and M. alba produced up to 39 mg L−1 using 70 mg L−1 p-coumaric acid. On the other hand, lines expressing STS from V. vinifera, A. hypogaea and R. tataricum generated resveratrol faster than other lines but accumulated lower amounts at the end of the batch period (27–30 mg L−1). The simultaneous consumption of ethanol and p-coumaric acid corroborates the role of ethanol as a carbon source involved in the conversion of p-coumaric acid into resveratrol.

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Separable functions of Tof1/Timeless in intra-S-checkpoint signalling, replisome stability and DNA topological stress

Nucleic Acids Research ◽

10.1093/nar/gkaa963 ◽

2020 ◽

Vol 48 (21) ◽

pp. 12169-12187

Author(s):

Rose Westhorpe ◽

Andrea Keszthelyi ◽

Nicola E Minchell ◽

David Jones ◽

Jonathan Baxter

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Replication ◽

Replication Stress ◽

Dna Helicase ◽

The Other ◽

Checkpoint Activation ◽

Replication Checkpoint ◽

Binding Partner ◽

Dna Replication Checkpoint

Abstract The highly conserved Tof1/Timeless proteins minimise replication stress and promote normal DNA replication. They are required to mediate the DNA replication checkpoint (DRC), the stable pausing of forks at protein fork blocks, the coupling of DNA helicase and polymerase functions during replication stress (RS) and the preferential resolution of DNA topological stress ahead of the fork. Here we demonstrate that the roles of the Saccharomyces cerevisiae Timeless protein Tof1 in DRC signalling and resolution of DNA topological stress require distinct N and C terminal regions of the protein, whereas the other functions of Tof1 are closely linked to the stable interaction between Tof1 and its constitutive binding partner Csm3/Tipin. By separating the role of Tof1 in DRC from fork stabilisation and coupling, we show that Tof1 has distinct activities in checkpoint activation and replisome stability to ensure the viable completion of DNA replication following replication stress.

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A Role for theSaccharomyces cerevisiaeRENT Complex Protein Net1 inHMRSilencing

Genetics ◽

10.1093/genetics/161.4.1411 ◽

2002 ◽

Vol 161 (4) ◽

pp. 1411-1423

Author(s):

Daniela Kasulke ◽

Stefanie Seitz ◽

Ann E Ehrenhofer-Murray

Keyword(s):

Saccharomyces Cerevisiae ◽

Protein A ◽

Rdna Locus ◽

The Other ◽

Central Component ◽

Yeast Saccharomyces Cerevisiae ◽

Sir Proteins ◽

Complex Protein ◽

Subtelomeric Regions

AbstractSilencing in the yeast Saccharomyces cerevisiae is known in three classes of loci: in the silent mating-type loci HML and HMR, in subtelomeric regions, and in the highly repetitive rDNA locus, which resides in the nucleolus. rDNA silencing differs markedly from the other two classes of silencing in that it requires a DNA-associated protein complex termed RENT. The Net1 protein, a central component of RENT, is required for nucleolar integrity and the control of exit from mitosis. Another RENT component is the NAD+-dependent histone deacetylase Sir2, which is the only silencing factor known to be shared among the three classes of silencing. Here, we investigated the role of Net1 in HMR silencing. The mutation net1-1, as well as NET1 expression from a 2μ-plasmid, restored repression at silencing-defective HMR loci. Both effects were strictly dependent on the Sir proteins. We found overexpressed Net1 protein to be directly associated with the HMR-E silencer, suggesting that Net1 could interact with silencer binding proteins and recruit other silencing factors to the silencer. In agreement with this, Net1 provided ORC-dependent, Sir1-independent silencing when artificially tethered to the silencer. In contrast, our data suggested that net1-1 acted indirectly in HMR silencing by releasing Sir2 from the nucleolus, thus shifting the internal competition for Sir2 from the silenced loci toward HMR.

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The Role of Nuclear Cap Binding Protein Cbc1p of Yeast in mRNA Termination and Degradation

Molecular and Cellular Biology ◽

10.1128/mcb.20.8.2827-2838.2000 ◽

2000 ◽

Vol 20 (8) ◽

pp. 2827-2838 ◽

Cited By ~ 48

Author(s):

Biswadip Das ◽

Zijian Guo ◽

Patrick Russo ◽

Pascal Chartrand ◽

Fred Sherman

Keyword(s):

Saccharomyces Cerevisiae ◽

Mrna Export ◽

Degradation Pathway ◽

The Other ◽

Nonsense Mutations ◽

Mrna Surveillance ◽

Nonessential Gene ◽

Cap Binding Complex ◽

Low Levels

ABSTRACT The cyc1-512 mutation in Saccharomyces cerevisiae causes a 90% reduction in the level of iso-1-cytochrome c because of the lack of a proper 3′-end-forming signal, resulting in low levels of eight aberrantly longcyc1-512 mRNAs which differ in length at their 3′ termini. cyc1-512 can be suppressed by deletion of either of the nonessential genes CBC1 and CBC2, which encode the CBP80 and CBP20 subunits of the nuclear cap binding complex, respectively, or by deletion of the nonessential gene UPF1, which encodes a major component of the mRNA surveillance complex. The upf1-Δ deletion suppressed the cyc1-512defect by diminishing degradation of the longer subset ofcyc1-512 mRNAs, suggesting that downstream elements or structures occurred in the extended 3′ region, similar to the downstream elements exposed by transcripts bearing premature nonsense mutations. On the other hand, suppression of cyc1-512defects by cbc1-Δ occurred by two different mechanisms. The levels of the shorter cyc1-512 transcripts were enhanced in the cbc1-Δ mutants by promoting 3′-end formation at otherwise-weak sites, whereas the levels of the longercyc1-512 transcripts, as well as of all mRNAs, were slightly enhanced by diminishing degradation. Furthermore,cbc1-Δ greatly suppressed the degradation of mRNAs and other phenotypes of a rat7-1 strain which is defective in mRNA export. We suggest that Cbc1p defines a novel degradation pathway that acts on mRNAs partially retained in nuclei.

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The origin of Brettanomyces bruxellensis in wines: a review

OENO One ◽

10.20870/oeno-one.2007.41.3.846 ◽

2007 ◽

Vol 41 (3) ◽

pp. 161 ◽

Cited By ~ 2

Author(s):

Vincent Renouf ◽

Aline Lonvaud-Funel ◽

Joana Coulon

Keyword(s):

Saccharomyces Cerevisiae ◽

Red Wine ◽

Microbial Consortium ◽

Yeast Species ◽

The Other ◽

Volatile Phenols ◽

Brettanomyces Bruxellensis ◽

Microbial Analysis ◽

Key Steps

Aims: This work reviews the latest knowledge concerning the role of Brettanomyces bruxellensis in red wine alteration.Results and conclusion: The origin of this yeast species and its place in the wine microbial consortium are discussed as well as microbial equilibriums with the other species, notably Saccharomyces cerevisiae and lactic acid bacteria. As a consequence, fermentations are described as key steps in Brettanomyces development management. Furthermore, the influence of ageing through the use of traditional winemaking practices is explained.Significance and impact of study: Finally, this paper emphases the need for a better understanding of chemical and microbial analysis together in order to better control this undesirable yeast and prevent the production of volatile phenols.

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Recombinant Saccharomyces cerevisiae Strain Expressing a Model Cytochrome P450 in the Rat Digestive Environment: Viability and Bioconversion Activity

Applied and Environmental Microbiology ◽

10.1128/aem.02091-06 ◽

2007 ◽

Vol 73 (11) ◽

pp. 3566-3574 ◽

Cited By ~ 10

Author(s):

G. Garrait ◽

J. F. Jarrige ◽

S. Blanquet ◽

E. Beyssac ◽

M. Alric

Keyword(s):

Saccharomyces Cerevisiae ◽

Drug Delivery ◽

Oral Administration ◽

Digestive Tract ◽

Drug Delivery Systems ◽

Recombinant Yeast ◽

Coumaric Acid ◽

Recombinant Microorganisms ◽

Small Intestinal

ABSTRACT An innovative “biodrug” concept, based on the oral administration of living recombinant microorganisms, has recently emerged for the prevention or treatment of various diseases. An engineered Saccharomyces cerevisiae strain expressing plant P450 73A1 (cinnamate-4-hydroxylase [CA4H] activity) was used, and its survival and ability to convert trans-cinnamic acid (CIN) into p-coumaric acid (COU) were investigated in vivo. In rats, the recombinant yeast was resistant to gastric and small intestinal secretions but was more sensitive to the conditions found in the large intestine. After oral administration of yeast and CIN, the CA4H activity was shown in vivo, with COU being found throughout the rat's digestive tract and in its urine. The bioconversion reaction occurred very fast, with most of the COU being produced within the first 5 min. The gastrointestinal sac technique demonstrated that the recombinant yeast was able to convert CIN into COU (conversion rate ranging from 2 to 5%) in all the organs of the rat's digestive tract: stomach, duodenum, jejunum, ileum, cecum, and colon. These results promise new opportunities for the development of drug delivery systems based on engineered yeasts catalyzing a bioconversion reaction directly in the digestive tract.

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Role of magnesium ions on Improvement of Fermentative activity of local yeast isolates Saccharomyces cerevisiae for ethanol production

Journal of Biotechnology Research Center ◽

10.24126/jobrc.2013.7.3.275 ◽

2013 ◽

Vol 7 (3) ◽

pp. 18-26

Author(s):

F. J. Shalesh ◽

F. R. Ali ◽

S. R. Frahan ◽

S. A. Mahmood ◽

A. M. Mari

Keyword(s):

Saccharomyces Cerevisiae ◽

Batch Fermentation ◽

Fossil Fuels ◽

Incubation Temperature ◽

Good Alternative ◽

Magnesium Ions ◽

Yeast Saccharomyces Cerevisiae ◽

Fermentative Activity ◽

Time Required

The interest in bio-ethanol increase as biofuel, it seems to be a good alternative for fossil fuels. The growth of yeast Saccharomyces cerevisiae is influence in micronutrient like ions. This research aims to evaluate the influence different amount of magnesium as co-enzyme in growth and fermentation of yeast S. cerevisiae which isolated from different local sources. The batch fermentation process was followed at different magnesium concentrations as magnesium chloride at incubation temperature 30C○ and pH 4.5, best supplement concentration was 1.25 gm/l by measuring the mass of the released carbon dioxide. Supplement media with magnesium prolonged exponential growth, reduced the decline in fermentation activity resulting in reduced the time required for the conversion glucose into ethanol.

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Separable functions of Tof1/Timeless in intra-S-checkpoint signalling, replisome stability and DNA topological stress

10.1101/2019.12.17.877811 ◽

2019 ◽

Author(s):

Rose Westhorpe ◽

Andrea Keszthelyi ◽

Nicola E. Minchell ◽

David Jones ◽

Jonathan Baxter

Keyword(s):

Saccharomyces Cerevisiae ◽

Dna Replication ◽

Replication Stress ◽

Dna Helicase ◽

The Other ◽

Checkpoint Activation ◽

Replication Checkpoint ◽

Binding Partner ◽

Dna Replication Checkpoint

AbstractThe highly conserved Tof1/Timeless proteins minimise replication stress and promote normal DNA replication. They are required to mediate the DNA replication checkpoint (DRC), the stable pausing of forks at protein fork blocks, the coupling of DNA helicase and polymerase functions during replication stress (RS) and the preferential resolution of DNA topological stress ahead of the fork. Here we demonstrate that the roles of the Saccharomyces cerevisiae Timeless protein Tof1 in DRC signalling and resolution of DNA topological stress require distinct N and C terminal regions of the protein, whereas the other functions of Tof1 are closely linked to the stable interaction between Tof1 and its constitutive binding partner Csm3/Tipin. By separating the role of Tof1 in DRC from fork stabilisation and coupling, we show that Tof1 has distinct activities in checkpoint activation and replisome stability to ensure the viable completion of DNA replication following replication stress.

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Potassium and Rubidium Effluxes in Saccharomyces cerevisiae

Zeitschrift für Naturforschung C ◽

10.1515/znc-1985-9-1024 ◽

1985 ◽

Vol 40 (9-10) ◽

pp. 721-725 ◽

Cited By ~ 15

Author(s):

María D . Ortega ◽

Alonso Rodríguez-Navarro

Keyword(s):

Saccharomyces Cerevisiae ◽

Ph Regulation ◽

Uptake System ◽

Acidic Ph ◽

Efflux System ◽

First Order ◽

Ph Values ◽

First Order Kinetics

Abstract In growing cells, K+ and Rb+ effluxes followed first order kinetics, took place with slight differences between K+ and Rb+ and were stimulated at acidic pH values. In uncoupled cells and ATP-depleted cells, K+ efflux was higher than Rb+ efflux. In ATP-depleted cells, the effluxes were not stimulated at acidic pH values. Two different K+ effluxes are proposed: (i) the normal efflux, which may be a K+/H+ antiport and does not function in ATP-depleted cells, and (ii) the backward reaction of the uptake system, which can be high in depolarized cells. The role of the normal efflux system in K+ content and pH regulation is discussed.

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Analysis of Spt7 Function in the Saccharomyces cerevisiae SAGA Coactivator Complex

Molecular and Cellular Biology ◽

10.1128/mcb.22.15.5367-5379.2002 ◽

2002 ◽

Vol 22 (15) ◽

pp. 5367-5379 ◽

Cited By ~ 90

Author(s):

Pei-Yun Jenny Wu ◽

Fred Winston

Keyword(s):

Saccharomyces Cerevisiae ◽

The Other ◽

Deletion Mutants ◽

Saga Complex ◽

Number Of Genes ◽

Processed Form ◽

Altered Form ◽

And Function ◽

Coactivator Complex

ABSTRACT The Saccharomyces cerevisiae SAGA complex is required for the normal transcription of a large number of genes. Complex integrity depends on three core subunits, Spt7, Spt20, and Ada1. We have investigated the role of Spt7 in the assembly and function of SAGA. Our results show that Spt7 is important in controlling the levels of the other core subunits and therefore of SAGA. In addition, partial SAGA complexes containing Spt7 can be assembled in the absence of both Spt20 and Ada1. Through biochemical and genetic analyses of a series of spt7 deletion mutants, we have identified a region of Spt7 required for interaction with the SAGA component Spt8. An adjacent Spt7 domain was found to be required for a processed form of Spt7 that is present in a previously identified altered form of SAGA called SLIK, SAGAalt, or SALSA. Analysis of an spt7 mutant with greatly reduced levels of SLIK/SAGAalt/SALSA suggests a subtle role for this complex in transcription that may be redundant with a subset of SAGA functions.

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Tryptophan Derivatives by Saccharomyces cerevisiae EC1118: Evaluation, Optimization, and Production in a Soybean-Based Medium

International Journal of Molecular Sciences ◽

10.3390/ijms22010472 ◽

2021 ◽

Vol 22 (1) ◽

pp. 472

Author(s):

Michele Dei Cas ◽

Ileana Vigentini ◽

Sara Vitalini ◽

Antonella Laganaro ◽

Marcello Iriti ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Kynurenic Acid ◽

Potential Role ◽

Defined Medium ◽

The Other ◽

Chemically Defined Medium ◽

Cultural Medium ◽

Whole Cell Biotransformation ◽

Tryptophan Derivatives

Given the pharmacological properti es and the potential role of kynurenic acid (KYNA) in human physiology and the pleiotropic activity of the neurohormone melatonin (MEL) involved in physiological and immunological functions and as regulator of antioxidant enzymes, this study aimed at evaluating the capability of Saccharomyces cerevisiae EC1118 to release tryptophan derivatives (dTRPs) from the kynurenine (KYN) and melatonin pathways. The setting up of the spectroscopic and chromatographic conditions for the quantification of the dTRPs in LC-MS/MS system, the optimization of dTRPs’ production in fermentative and whole-cell biotransformation approaches and the production of dTRPs in a soybean-based cultural medium naturally enriched in tryptophan, as a case of study, were included in the experimental plan. Variable amounts of dTRPs, with a prevalence of metabolites of the KYN pathway, were detected. The LC-MS/MS analysis showed that the compound synthesized at highest concentration is KYNA that reached 9.146 ± 0.585 mg/L in fermentation trials in a chemically defined medium at 400 mg/L TRP. Further experiments in a soybean-based medium confirm KYNA as the main dTRPs, whereas the other dTRPs reached very lower concentrations. While detectable quantities of melatonin were never observed, two MEL isomers were successfully measured in laboratory media.

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