Characterization of non-oxidative transaldolase and transketolase enzymes in the pentose phosphate pathway with regard to xylose utilization by recombinant Saccharomyces cerevisiae

Matsushika, Akinori; Goshima, Tetsuya; Fujii, Tatsuya; Inoue, Hiroyuki; Sawayama, Shigeki; Yano, Shinichi

doi:10.1016/j.enzmictec.2012.03.008

Cited by 67 publications

(42 citation statements)

References 42 publications

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“…Our present finding that the tkl1/2⌬ tal1⌬ strain displayed decreased fermentation rates (Fig. 4B), together with the findings from previous reports that deletion of the TKL1 gene alone is sufficient to reduce glucose utilization (48,49), suggests that Tkl1p plays a positive role in glycolysis and/or alcoholic fermentation.…”

Section: Discussionsupporting

confidence: 75%

Inhibitory Role of Greatwall-Like Protein Kinase Rim15p in Alcoholic Fermentation via Upregulating the UDP-Glucose Synthesis Pathway in Saccharomyces cerevisiae

Watanabe

Zhou

Hirata

et al. 2016

Appl Environ Microbiol

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The high fermentation rate of Saccharomyces cerevisiae sake yeast strains is attributable to a loss-of-function mutation in the RIM15 gene, which encodes a Greatwall-family protein kinase that is conserved among eukaryotes. In the present study, we performed intracellular metabolic profiling analysis and revealed that deletion of the RIM15 gene in a laboratory strain impaired glucose-anabolic pathways through the synthesis of UDP-glucose (UDPG). Although Rim15p is required for the synthesis of trehalose and glycogen from UDPG upon entry of cells into the quiescent state, we found that Rim15p is also essential for the accumulation of cell wall ␤-glucans, which are also anabolic products of UDPG. Furthermore, the impairment of UDPG or 1,3-␤-glucan synthesis contributed to an increase in the fermentation rate. Transcriptional induction of PGM2 (phosphoglucomutase) and UGP1 (UDPG pyrophosphorylase) was impaired in Rim15p-deficient cells in the early stage of fermentation. These findings demonstrate that the decreased anabolism of glucose into UDPG and 1,3-␤-glucan triggered by a defect in the Rim15p-mediated upregulation of PGM2 and UGP1 redirects the glucose flux into glycolysis. Consistent with this, sake yeast strains with defective Rim15p exhibited impaired expression of PGM2 and UGP1 and decreased levels of ␤-glucans, trehalose, and glycogen during sake fermentation. We also identified a sake yeast-specific mutation in the glycogen synthesis-associated glycogenin gene GLG2, supporting the conclusion that the glucose-anabolic pathway is impaired in sake yeast. These findings demonstrate that downregulation of the UDPG synthesis pathway is a key mechanism accelerating alcoholic fermentation in industrially utilized S. cerevisiae sake strains. S ake yeast strains, which belong to the species Saccharomyces cerevisiae, are capable of achieving ethanol yields as high as 22 vol% in fermenting sake mash (1-3). This characteristic phenotype is attributed in part to their high and sustained maximum fermentation rates, as observed in batch cultures containing high concentrations of glucose (4), and is due to the continuous supply of fermentable sugars to yeast cells in sake mash via the degradation of rice starch by enzymes produced by Aspergillus oryzae. In recent studies of the representative sake yeast strain Kyokai no. 7 (K7) and its relatives, we revealed that several stress-and/or nutrient-responsive transcription factors, particularly Msn2p and Msn4p (Msn2/4p), Hsf1p, Adr1p, and Cat8p, are significantly inactivated (5-7). Impairment of these transcriptional activators in laboratory strains of S. cerevisiae leads to increased fermentation rates, indicating that defective stress responses are linked with the superior fermentation properties of sake yeast (2, 5-7). Moreover, a loss-of-function mutation by insertion of an A residue at position 5055 in the RIM15 gene (rim15 5055insA ) was commonly found among K7-related strains (8). RIM15 encodes a conserved Greatwall-like protein kinase involved in the control of mitot...

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Section: Discussionsupporting

confidence: 75%

Inhibitory Role of Greatwall-Like Protein Kinase Rim15p in Alcoholic Fermentation via Upregulating the UDP-Glucose Synthesis Pathway in Saccharomyces cerevisiae

Watanabe

Zhou

Hirata

et al. 2016

Appl Environ Microbiol

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“…Among up-regulated genes were ZWF1 encoding glucose-6-phosphate dehydrogenase, which catalyzes the first and rate-limiting step of PPP [52], and TKL2 encoding transketolase [53]. PPP with its oxidative branch is a major source of reducing equivalents (NADPH) and metabolic intermediates (e.g., ribose 5-phosphate and erythrose 4-phosphate) required for biosynthetic processes [54], thereby playing an important role in the resistance to oxidative stress [55].…”

Section: Resultsmentioning

confidence: 99%

Transcriptional and antioxidative responses to endogenous polyunsaturated fatty acid accumulation in yeast

et al. 2014

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Pathophysiology of polyunsaturated fatty acids (PUFAs) is associated with aberrant lipid and oxygen metabolism. In particular, under oxidative stress, PUFAs are prone to autocatalytic degradation via peroxidation, leading to formation of reactive aldehydes with numerous potentially harmful effects. However, the pathological and compensatory mechanisms induced by lipid peroxidation are very complex and not sufficiently understood. In our study, we have used yeast capable of endogenous PUFA synthesis in order to understand the effects triggered by PUFA accumulation on cellular physiology of a eukaryotic organism. The mechanisms induced by PUFA accumulation in S. cerevisiae expressing Hevea brasiliensis Δ12-fatty acid desaturase include down-regulation of components of electron transport chain in mitochondria as well as up-regulation of pentose-phosphate pathway and fatty acid β-oxidation at the transcriptional level. Interestingly, while no changes were observed at the transcriptional level, activities of two important enzymatic antioxidants, catalase and glutathione-S-transferase, were altered in response to PUFA accumulation. Increased intracellular glutathione levels further suggest an endogenous oxidative stress and activation of antioxidative defense mechanisms under conditions of PUFA accumulation. Finally, our data suggest that PUFA in cell membrane causes metabolic changes which in turn lead to adaptation to endogenous oxidative stress.

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“…Further indications of shifts in the transgenics’ carbon metabolism include the following: Fructose-bisphosphate aldolase (POPTR_0006s17940.1) and glyceraldehyde-3-phosphate dehydrogenase (POPTR_0015s10330.2, GAPDH 1.2) was upregulated at the transcript but downregulated at the protein level (POPTR_0010s06560.1 GAPDH.3, POPTR_0012s09570.1 GAPDH 1.1; Additional file 2: Table S1.2, Additional file 2: Table S2.2; Table 1). Transaldolase (POPTR_0003s16030.1) and transketolase (POPTR_0002s14730.1), both of which provide reversible links between the PPP and glycolysis [66], were upregulated at transcript and protein levels respectively. PPP and glycolysis have been suggested to contribute to ROS balance and scavenging [67-69].…”

Section: Resultsmentioning

confidence: 99%

OnPLS integration of transcriptomic, proteomic and metabolomic data shows multi-level oxidative stress responses in the cambium of transgenic hipI- superoxide dismutase Populus plants

et al. 2013

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BackgroundReactive oxygen species (ROS) are involved in the regulation of diverse physiological processes in plants, including various biotic and abiotic stress responses. Thus, oxidative stress tolerance mechanisms in plants are complex, and diverse responses at multiple levels need to be characterized in order to understand them. Here we present system responses to oxidative stress in Populus by integrating data from analyses of the cambial region of wild-type controls and plants expressing high-isoelectric-point superoxide dismutase (hipI-SOD) transcripts in antisense orientation showing a higher production of superoxide. The cambium, a thin cell layer, generates cells that differentiate to form either phloem or xylem and is hypothesized to be a major reason for phenotypic perturbations in the transgenic plants. Data from multiple platforms including transcriptomics (microarray analysis), proteomics (UPLC/QTOF-MS), and metabolomics (GC-TOF/MS, UPLC/MS, and UHPLC-LTQ/MS) were integrated using the most recent development of orthogonal projections to latent structures called OnPLS. OnPLS is a symmetrical multi-block method that does not depend on the order of analysis when more than two blocks are analysed. Significantly affected genes, proteins and metabolites were then visualized in painted pathway diagrams.ResultsThe main categories that appear to be significantly influenced in the transgenic plants were pathways related to redox regulation, carbon metabolism and protein degradation, e.g. the glycolysis and pentose phosphate pathways (PPP). The results provide system-level information on ROS metabolism and responses to oxidative stress, and indicate that some initial responses to oxidative stress may share common pathways.ConclusionThe proposed data evaluation strategy shows an efficient way of compiling complex, multi-platform datasets to obtain significant biological information.

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Characterization of non-oxidative transaldolase and transketolase enzymes in the pentose phosphate pathway with regard to xylose utilization by recombinant Saccharomyces cerevisiae

Cited by 67 publications

References 42 publications

Inhibitory Role of Greatwall-Like Protein Kinase Rim15p in Alcoholic Fermentation via Upregulating the UDP-Glucose Synthesis Pathway in Saccharomyces cerevisiae

Inhibitory Role of Greatwall-Like Protein Kinase Rim15p in Alcoholic Fermentation via Upregulating the UDP-Glucose Synthesis Pathway in Saccharomyces cerevisiae

Transcriptional and antioxidative responses to endogenous polyunsaturated fatty acid accumulation in yeast

OnPLS integration of transcriptomic, proteomic and metabolomic data shows multi-level oxidative stress responses in the cambium of transgenic hipI- superoxide dismutase Populus plants

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