Dataset Information

Effects of physicochemical parameters on volatile sulphur compound formation from L-methionine catabolism by non-growing cells of Kluyveromyces lactis.

ABSTRACT: The present study investigated for the first time the effects of various physicochemical parameters on the production of volatile sulphur compounds (VSCs) by non-growing cells of Kluyveromyces lactis supplemented with L-methionine. The results showed that the production of VSCs positively correlated with the cell biomass, but it seemed that no clear relationship with L-methionine concentration existed. Temperature and pH significantly affected the formation of VSCs with more production at 30 °C and pH 5, respectively. Nitrogen supplementation (in the form of diammonium phosphate, DAP) repressed the production of VSCs. It is interesting to note that DAP and yeast extract supplementation induced the production of methional, but not Mn²⁺ supplementation. The presence of Mn²⁺ improved the production of methionol and dimethyl disulphide, but inhibited the formation of S-methyl thioacetate. The study indicated that optimization of physicochemical conditions and media composition would be crucial for producing L-methionine-derived VSC bioflavor.

SUBMITTER: Lu Y

PROVIDER: S-EPMC6029988 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Effects of physicochemical parameters on volatile sulphur compound formation from L-methionine catabolism by non-growing cells of Kluyveromyces lactis.

Lu Yuyun Y Nawrath Margarete M Sun Jingcan J Liu Shao-Quan SQ

AMB Express 20180703 1

The present study investigated for the first time the effects of various physicochemical parameters on the production of volatile sulphur compounds (VSCs) by non-growing cells of Kluyveromyces lactis supplemented with L-methionine. The results showed that the production of VSCs positively correlated with the cell biomass, but it seemed that no clear relationship with L-methionine concentration existed. Temperature and pH significantly affected the formation of VSCs with more production at 30 °C ...[more]

PMID: 29971574

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Project description:Adaptation to environmental perturbations requires living systems to coordinately regulate signaling pathways, gene expression, and metabolism. To better understand the mechanisms underlying adaptation, the regulatory nodes within networks must be elucidated. Here, ARO8-2 (which encodes an aminotransferase), PDC (which encodes a decarboxylase), and STR3 (which encodes a demethiolase) were identified as key genes involved in the catabolism of methionine in the mycoparasitic fungus Clonostachys rosea, isolated from Tuber melanosporum ascocarps. Exogenous Met induced the transcription of ARO8-2 and PDC but repressed the transcription of STR3, which is controlled by the putative MSN2 and GLN3 binding sites responding to nitrogen catabolite repression. Met and its structural derivatives function as glutamine synthetase inhibitors, resulting in the downregulation of STR3 expression. The putative GLN3 binding site was necessary for STR3 downregulation. In Saccharomyces cerevisiae, Met and its structural derivatives also triggered downregulation of demethiolase gene expression. Altogether, the results indicated that exogenous Met triggered nitrogen catabolite repression, which stimulated the Ehrlich pathway and negatively regulated the demethiolation pathway via the methionine sulfoximine-responsive regulatory pathway. This finding revealed the regulatory nodes within the networks controlling the catabolism of Met into volatile organic sulfur-containing compounds, thereby enhancing our understanding of adaptation.IMPORTANCE Methionine shuttles organic nitrogen and plays a central role in nitrogen metabolism. Exogenous Met strongly induces the expression of ARO8-2 and PDC, represses the expression of STR3, and generates volatile organic sulfur-containing compounds via the Ehrlich and demethiolation pathways. In this study, we used genetic, bioinformatic, and metabolite-based analyses to confirm that transcriptional control of the aminotransferase gene ARO8-2, the decarboxylase gene PDC, and the demethiolase gene STR3 modulates Met catabolism into volatile organic sulfur-containing compounds. Importantly, we found that, in addition to the Ehrlich pathway, the demethiolation pathway was regulated by a nitrogen catabolite repression-sensitive regulatory pathway that controlled the transcription of genes required to catabolize poor nitrogen sources. This work significantly advances our understanding of nitrogen catabolite repression-sensitive transcriptional regulation of sulfur-containing amino acid catabolism and provides a basis for engineering Met catabolism pathways for the production of fuel and valuable flavor alcohols.

Project description:BackgroundEven before having its genome sequence published in 2004, Kluyveromyces lactis had long been considered a model organism for studies in genetics and physiology. Research on Kluyveromyces lactis is quite advanced and this yeast species is one of the few with which it is possible to perform formal genetic analysis. Nevertheless, until now, no complete metabolic functional annotation has been performed to the proteins encoded in the Kluyveromyces lactis genome.ResultsIn this work, a new metabolic genome-wide functional re-annotation of the proteins encoded in the Kluyveromyces lactis genome was performed, resulting in the annotation of 1759 genes with metabolic functions, and the development of a methodology supported by merlin (software developed in-house). The new annotation includes novelties, such as the assignment of transporter superfamily numbers to genes identified as transporter proteins. Thus, the genes annotated with metabolic functions could be exclusively enzymatic (1410 genes), transporter proteins encoding genes (301 genes) or have both metabolic activities (48 genes). The new annotation produced by this work largely surpassed the Kluyveromyces lactis currently available annotations. A comparison with KEGG's annotation revealed a match with 844 (~90%) of the genes annotated by KEGG, while adding 850 new gene annotations. Moreover, there are 32 genes with annotations different from KEGG.ConclusionsThe methodology developed throughout this work can be used to re-annotate any yeast or, with a little tweak of the reference organism, the proteins encoded in any sequenced genome. The new annotation provided by this study offers basic knowledge which might be useful for the scientific community working on this model yeast, because new functions have been identified for the so-called metabolic genes. Furthermore, it served as the basis for the reconstruction of a compartmentalized, genome-scale metabolic model of Kluyveromyces lactis, which is currently being finished.

Dataset Information

Effects of physicochemical parameters on volatile sulphur compound formation from L-methionine catabolism by non-growing cells of Kluyveromyces lactis.

Publications

Effects of physicochemical parameters on volatile sulphur compound formation from L-methionine catabolism by non-growing cells of Kluyveromyces lactis.

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