RRC ID 55840
Author Masumoto H, Matsuyama S.
Title The combination of NAD+-dependent deacetylase gene deletion and the interruption of gluconeogenesis causes increased glucose metabolism in budding yeast.
Journal PLoS One
Abstract Metabolic engineering focuses on rewriting the metabolism of cells to enhance native products or endow cells with the ability to produce new products. This engineering has the potential for wide-range application, including the production of fuels, chemicals, foods and pharmaceuticals. Glycolysis manages the levels of various secondary metabolites by controlling the supply of glycolytic metabolites. Metabolic reprogramming of glycolysis is expected to cause an increase in the secondary metabolites of interest. In this study, we constructed a budding yeast strain harboring the combination of triple sirtuin gene deletion (hst3∆ hst4∆ sir2∆) and interruption of gluconeogenesis by the deletion of the FBP1 gene encoding fructose-1,6-bisphosphatase (fbp1∆). hst3∆ hst4∆ sir2∆ fbp1∆ cells harbored active glycolysis with high glucose consumption and active ethanol productivity. Using capillary electrophoresis-time-of-flight mass spectrometry (CE-TOF/MS) analysis, hst3∆ hst4∆ sir2∆ fbp1∆ cells accumulated not only glycolytic metabolites but also secondary metabolites, including nucleotides that were synthesized throughout the pentose phosphate (PP) pathway, although various amino acids remained at low levels. Using the stable isotope labeling assay for metabolites, we confirmed that hst3∆ hst4∆ sir2∆ fbp1∆ cells directed the metabolic fluxes of glycolytic metabolites into the PP pathway. Thus, the deletion of three sirtuin genes (HST3, HST4 and SIR2) and the FBP1 gene can allow metabolic reprogramming to increase glycolytic metabolites and several secondary metabolites except for several amino acids.
Volume 13(3)
Pages e0194942
Published 2018-3-26
DOI 10.1371/journal.pone.0194942
PII PONE-D-17-34706
PMID 29579121
PMC PMC5868833
MeSH Carbon Isotopes / chemistry Electrophoresis, Capillary Fructose-Bisphosphatase / genetics* Fructose-Bisphosphatase / metabolism Gluconeogenesis / genetics* Glucose / analysis Glucose / metabolism* Glycolysis Histone Deacetylases / deficiency Histone Deacetylases / genetics* Isotope Labeling Mass Spectrometry Metabolic Engineering* Metabolome Nucleotides / analysis Nucleotides / metabolism Pentose Phosphate Pathway / physiology Principal Component Analysis Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae / growth & development Saccharomyces cerevisiae / metabolism* Saccharomyces cerevisiae Proteins / genetics* Saccharomyces cerevisiae Proteins / metabolism Silent Information Regulator Proteins, Saccharomyces cerevisiae / deficiency Silent Information Regulator Proteins, Saccharomyces cerevisiae / genetics* Sirtuin 2 / deficiency Sirtuin 2 / genetics*
IF 2.74
Times Cited 2
Prokaryotes E. coli JM109