Genetic manipulation of HSP26 and YHR087W stress genes may improve fermentative behaviour in wine yeasts under vinification conditions

Jiménez‐Martí, Elena; Zuzuarregui, Aurora; Ridaura, I.; Lozano, Noelia; Olmo, Marcel·lí del

doi:10.1016/j.ijfoodmicro.2009.01.017

Cited by 21 publications

(10 citation statements)

References 29 publications

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“…In order to improve wine quality as well as the elaboration processes, wine yeast strains have been selected for a very specialized phenotype and are genetically complex, usually being polyploid or aneuploid (6). To elucidate whether the better ability to convert endogenous GPP into monoterpenes is a common feature in oenological strains of S. cerevisiae, two different wine strains, T 73 -4 (nearly diploid [37], derived from T 73 Lallemand) and ICV16ura3⌬ (derived from ICV16 Fermicru Primeur, DSM [25]), as well as the laboratory diploid strain FY1679 (MATa/MAT␣ ura3-52/ura3-52; isogenic to BQS252 and BQS253), were transformed with YEplac195-Lis and two uracil-prototrophic transformants of each genetic background were isolated. Figure 2A shows that linalool production by recombinant FY1679Lis strains (YR201 and YR202) was not greater than that of recombinant BQS252Lis and BQS253Lis, indicating that diploidy alone does not improve linalool levels.…”

Section: Resultsmentioning

confidence: 99%

Enhanced Production of a Plant Monoterpene by Overexpression of the 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Catalytic Domain in Saccharomyces cerevisiae

Rico

Pardo

Orejas

2010

Appl Environ Microbiol

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Linalool production was evaluated in different Saccharomyces cerevisiae strains expressing the Clarkia breweri linalool synthase gene (LIS). The wine strain T 73 was shown to produce higher levels of linalool than conventional laboratory strains (i.e., almost three times the amount). The performance of this strain was further enhanced by manipulating the endogenous mevalonate (MVA) pathway: deregulated overexpression of the rate-limiting 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) doubled linalool production. In a haploid laboratory strain, engineering of this key step also improved linalool yield.Monoterpenes are a class of isoprenoids of increasing industrial and clinical interest usually produced by plants. They are used as aromatic additives in the food and cosmetics industries and are also important components in wine aroma. Moreover, certain monoterpenes display antimicrobial, antiparasitic, and antiviral properties as well as a plethora of promising health benefits (for recent reviews, see references 2, 7, 15, 28, and 30 and references cited therein). To date, many studies have focused on plant metabolic engineering of monoterpene production (for selected reviews, see references 1, 14, 19, 29, and 35 and references cited therein), and few studies have been carried out on microorganisms (9,21,22,34,38). Efficient microbial production of these metabolites could provide an alternative to the current methods of chemical synthesis or extraction from natural sources. In this regard, a considerable number of studies have shown the utility of Saccharomyces cerevisiae as a valuable platform for sesquiterpene, diterpene, triterpene, and carotene production (references 5, 10, 23, 26, 30, 31, 32, and 33 and references cited therein). However, all the efforts dedicated to the improvement of isoprenoid yields in S. cerevisiae have been performed using conventional laboratory strains, and there are no studies concerning natural or industrially relevant isolates.In recent years, many genes that encode plant monoterpene synthases (MTS), a family of enzymes which specifically catalyze the conversion of the ubiquitous C 10 intermediate of isoprenoid biosynthesis geranyl pyrophosphate (GPP) to monoterpenes, have been characterized. Such is the case with the LIS gene (codes for S-linalool synthase) of Clarkia breweri, the first MTS-encoding gene to be isolated (13). In contrast to plants, S. cerevisiae cannot produce monoterpenes efficiently, mainly due to the lack of specific pathways involving MTS. However, GPP is formed as a transitory intermediate in the two-step synthesis of farnesyl pyrophosphate (FPP), catalyzed by FPP synthase (FPPS) (Fig. 1), and some natural S. cerevisiae strains have been shown to possess the ability to produce small amounts of monoterpenes (8). Whether this occurs through unspecific dephosphorylation of a more available endogenous pool of GPP and subsequent bioconversions is not known. In addition, it has recently been established that S. cerevisiae has enough free GPP to...

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Section: Resultsmentioning

confidence: 99%

Enhanced Production of a Plant Monoterpene by Overexpression of the 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Catalytic Domain in Saccharomyces cerevisiae

Rico

Pardo

Orejas

2010

Appl Environ Microbiol

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“…The results previously obtained in our laboratory indicate an involvement of Yhr087wp in the osmotic stress caused by high glucose concentrations, although the gene expression data published by other authors suggest that it may also participate in the response to other forms of stress [2,11]. The link between Yhr087wp and response to stress (particularly osmotic stress caused by high glucose concentrations) is based on the following aspects: i) deletion of the coding gene results in a lower growth rate in the presence of 25% and 30% glucose concentrations [15] ii) wine yeast strains displaying better tolerance to osmotic stress have high levels of the mRNA of this gene [15]; iii) its overexpression improves the osmotic stress resistance and fermentative behavior of these strains [17]; iv) the proteomic study carried out with the Δyhr087w deletion mutant [15] provides a strong link between the levels of this gene expression under high glucose concentrations and the Hsp104p and Hsp78p levels; v) genetic interactions between YHR087W and HSP82 , another gene encoding an Hsp protein [29] have been described. This work demonstrates another interesting interaction which was previously suggested by Constanzo et al [28]: Δyhr087w is capable of partially suppressing the growth defects displayed by the Δbcy1 mutant at 37°C.…”

Section: Discussionmentioning

confidence: 99%

“…cDNA was obtained as described previously [47]. The absence of DNA contamination in these preparations was assessed by analyzing the intron-containing gene ACT1 by semi-quantitative RT-PCR using the oligonucleotides ACT-1 and ACT-2 shown in Additional file 1: Table S2 [17]. The ACT1 gene was also used as a reference gene in some experiments given its constitutive expression under the conditions considered in this work [2,15].…”

Section: Methodsmentioning

confidence: 99%

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The Saccharomyces cerevisiae Hot1p regulated gene YHR087W (HGI1) has a role in translation upon high glucose concentration stress

2012

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BackgroundWhile growing in natural environments yeasts can be affected by osmotic stress provoked by high glucose concentrations. The response to this adverse condition requires the HOG pathway and involves transcriptional and posttranscriptional mechanisms initiated by the phosphorylation of this protein, its translocation to the nucleus and activation of transcription factors. One of the genes induced to respond to this injury is YHR087W. It encodes for a protein structurally similar to the N-terminal region of human SBDS whose expression is also induced under other forms of stress and whose deletion determines growth defects at high glucose concentrations.ResultsIn this work we show that YHR087W expression is regulated by several transcription factors depending on the particular stress condition, and Hot1p is particularly relevant for the induction at high glucose concentrations. In this situation, Hot1p, together to Sko1p, binds to YHR087W promoter in a Hog1p-dependent manner. Several evidences obtained indicate Yhr087wp’s role in translation. Firstly, and according to TAP purification experiments, it interacts with proteins involved in translation initiation. Besides, its deletion mutant shows growth defects in the presence of translation inhibitors and displays a slightly slower translation recovery after applying high glucose stress than the wild type strain. Analyses of the association of mRNAs to polysome fractions reveals a lower translation in the mutant strain of the mRNAs corresponding to genes GPD1, HSP78 and HSP104.ConclusionsThe data demonstrates that expression of Yhr087wp under high glucose concentration is controlled by Hot1p and Sko1p transcription factors, which bind to its promoter. Yhr087wp has a role in translation, maybe in the control of the synthesis of several stress response proteins, which could explain the lower levels of some of these proteins found in previous proteomic analyses and the growth defects of the deletion strain.

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“…In S. cerevisiae, the deletion of HSP26 causes an abnormal cell shape and accumulation of protein aggregates under stress conditions, but does not affect thermotolerance (Franzmann et al, 2008). Also, the overexpression of HSP26 showed that the viability under particular stress conditions was not improved (Jiménez-Martí et al, 2009). Moreover, some HSP proteins work together as partners with co-chaperons or co-factors to resist adverse conditions, thus, the manipulation of one special gene was not effective for improving the stress resistance phenotype of yeast cells.…”

Section: Genes Involved In Heat Shock Protein (Hsp)mentioning

confidence: 99%

Transcriptome analysis of Saccharomyces cerevisiae at the late stage of very high gravity (VHG) fermentation

Zhang¹,

Zhao²,

Guo-hua³

et al. 2012

Afr. J. Biotechnol.

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DNA microarray analysis was used to investigate the expression profile of Saccharomyces cerevisiae genes in glycolysis pathway, trehalose and steroid biosynthesis and heat shock proteins (HSP) in response to harsh environment under the late stage of very high gravity (VHG) fermentation. The data show that only a few genes (GPM2, PGM1, GAL10 and PGM1) involved in glycolysis pathway and trehalose biosynthesis were up-regulated. Five genes that encode heat shock proteins (HSP26, HSP10, HSP42, HSP78 and HSP82) were up-regulated. Among these five genes, there was a strong expression increase of about 84-fold for HSP26. The results of this study revealed adverse VHG fermentation conditions stress response pattern and suggested interesting information about the mechanisms involved in adaptation of cells to the complex VHG fermentation environment. This identification of genes provides information that will help to genetically modify yeast to further maintain the fermentation fitness and improve its fermentation capacity and process.

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Genetic manipulation of HSP26 and YHR087W stress genes may improve fermentative behaviour in wine yeasts under vinification conditions

Cited by 21 publications

References 29 publications

Enhanced Production of a Plant Monoterpene by Overexpression of the 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Catalytic Domain in Saccharomyces cerevisiae

Enhanced Production of a Plant Monoterpene by Overexpression of the 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Catalytic Domain in Saccharomyces cerevisiae

The Saccharomyces cerevisiae Hot1p regulated gene YHR087W (HGI1) has a role in translation upon high glucose concentration stress

Transcriptome analysis of Saccharomyces cerevisiae at the late stage of very high gravity (VHG) fermentation

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