Inhibitory Role of Greatwall-Like Protein Kinase Rim15p in Alcoholic Fermentation via Upregulating the UDP-Glucose Synthesis Pathway in Saccharomyces cerevisiae.
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 β-glucans, which are also anabolic products of UDPG. Furthermore, the impairment of UDPG or 1,3-β-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-β-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 β-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.
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan National Research Institute of Brewing, Higashihiroshima, Hiroshima, Japan.
Daisuke
D
Watanabe
National Research Institute of Brewing, Higashihiroshima, Hiroshima, Japan.
Yan
Y
Zhou
Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba, Japan.
Aiko
A
Hirata
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
Yukiko
Y
Sugimoto
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
Kenichi
K
Takagi
National Research Institute of Brewing, Higashihiroshima, Hiroshima, Japan.
Takeshi
T
Akao
Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba, Japan.
Yoshikazu
Y
Ohya
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan.
Hiroshi
H
Takagi
National Research Institute of Brewing, Higashihiroshima, Hiroshima, Japan Faculty of Agriculture, Iwate University, Morioka, Iwate, Japan simoi@iwate-u.ac.jp.
Hitoshi
H
Shimoi
D000434:Alcoholic Beverages / Q000382:microbiology; D053898:Biosynthetic Pathways / Q000235:genetics* / Q000502:physiology; D002473:Cell Wall; D015536:Down-Regulation; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014199:Trehalose / Q000378:metabolism; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000235:genetics; D015854:Up-Regulation; D014532:Uridine Diphosphate Glucose / Q000096:biosynthesis; D047071:beta-Glucans / Q000378:metabolism
D029701:Saccharomyces cerevisiae Proteins; D047071:beta-Glucans; D000431:Ethanol; D006003:Glycogen; C033363:beta-1,3-glucan; D014199:Trehalose; D011494:Protein Kinases; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; D005947:Glucose; D014532:Uridine Diphosphate Glucose