Synthetic rescue couples NADPH generation to metabolite overproduction in Saccharomyces cerevisiae

Partow, Siavash; Hyland, Patrick; Mahadevan, Radhakrishnan

doi:10.1016/j.ymben.2017.08.004

Cited by 16 publications

(8 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3 ). In addition, the disruption of the PP pathway was previously reported to increase carbon fluxes through cytosolic aldehyde dehydrogenase in the acetate production pathway 11 , 41 . Therefore, we chose to delete ZWF1 for the elimination of the PP pathway.…”

Section: Resultsmentioning

confidence: 95%

Rerouting of NADPH synthetic pathways for increased protopanaxadiol production in Saccharomyces cerevisiae

Kim

Jang

Sung

et al. 2018

Sci Rep

View full text Add to dashboard Cite

Ginseng (Panax ginseng) and its bioactive components, ginsenosides, are popular medicinal herbal products, exhibiting various pharmacological effects. Despite their advocated use for medication, the long cultivation periods of ginseng roots and their low ginsenoside content prevent mass production of this compound. Yeast Saccharomyces cerevisiae was engineered for production of protopanaxadiol (PPD), a type of aglycone characterizing ginsenoside. PPD-producing yeast cell factory was further engineered by obtaining a balance between enzyme expressions and altering cofactor availability. Different combinations of promoters (PGPD, PCCW12, and PADH2) were utilized to construct the PPD biosynthetic pathway. Rerouting the redox metabolism to improve NADPH availability in the engineered S. cerevisiae also increased PPD production. Combining these approaches resulted in more than an 11-fold increase in PPD titer over the initially constructed strain. The series of metabolic engineering strategies of this study provides a feasible approach for the microbial production of PPD and development of microbial platforms producing other industrially-relevant terpenoids.

show abstract

Section: Resultsmentioning

confidence: 95%

Rerouting of NADPH synthetic pathways for increased protopanaxadiol production in Saccharomyces cerevisiae

Kim

Jang

Sung

et al. 2018

Sci Rep

View full text Add to dashboard Cite

show abstract

“…However, this enzyme level is elevated only when glucose in the medium is depleted in a diauxic shift or during growth on nonfermentable carbon sources, for example, glycerol or ethanol . Interruption of the cytosolic redox metabolism through deletion of ZWF1 results in an increased acetate and oxoglutarate production . In turn, the Δ zwf1 Δ idp2 double yeast mutant rapidly loses cell viability following shifts from medium containing glucose to medium containing either oleate or acetate as a carbon source …”

Section: Introductionmentioning

confidence: 99%

“…19 Interruption of the cytosolic redox metabolism through deletion of ZWF1 results in an increased acetate and oxoglutarate production. 20 In turn, the Δzwf1Δidp2 double yeast mutant rapidly loses cell viability following shifts from medium containing glucose to medium containing either oleate or acetate as a carbon source. 17 The aim of this study was to investigate the effect of the lack of enzymes responsible for NADPH production in the PP pathway, on changes in the cellular redox status and their implications on growth of cell population, the reproductive potential and total lifespan of the S. cerevisiae yeast.…”

mentioning

confidence: 99%

Disorders in NADPH generation via pentose phosphate pathway influence the reproductive potential of the Saccharomyces cerevisiae yeast due to changes in redox status

Kwolek‐Mirek

Maslanka

Mołoń

2018

J of Cellular Biochemistry

View full text Add to dashboard Cite

Intermediary metabolites have a crucial impact on basic cell functions. There is a relationship between cellular metabolism and redox balance. To maintain redox homoeostasis, the cooperation of both glutathione and nicotine adenine dinucleotides is necessary. Availability of nicotinamide adenine dinucleotide phosphate (NADPH) as a major electron donor is critical for many intracellular redox reactions. The activity of glucose‐6‐phosphate dehydrogenase (Zwf1p) and 6‐phosphogluconate dehydrogenase (Gnd1p and Gnd2p) is responsible for NADPH formation in a pentose phosphate (PP) pathway. In this study, we examine the impact of redox homoeostasis on cellular physiology and proliferation. We have noted that the Δzwf1 mutant lacking the rate‐limiting enzyme of the PP pathway shows changes in the cellular redox status caused by disorders in NADPH generation. This leads to a decrease in reproductive potential but without affecting the total lifespan of the cell. The results presented in this paper show that nicotine adenine dinucleotides play a central role in cellular physiology.

show abstract

“…Both mechanisms have the potential to globally remodel the metabolic capacity of a cell with limited overall genetic changes. Second, while high throughput studies have the advantage of being comprehensive, they require sifting through non-functional "rider" mutations [86], making true rescue mutations difficult to identify and leaving interactions undiscovered (as evidenced by a comparison with a targeted study [87]). When contrasted with large-scale studies in yeast, our small-scale forward genetic study in E. coli offers higher sensitivity, enhanced targeting of specific genes, and broader applicability.…”

Section: Toward Systematic Discovery Of Synthetic Rescue Interactionsmentioning

confidence: 99%

Experimental evolution of diverse Escherichia coli metabolic mutants identifies genetic loci for convergent adaptation of growth rate

Wytock¹,

Fiebig²,

Willett³

et al. 2018

PLoS Genet

View full text Add to dashboard Cite

Cell growth is determined by substrate availability and the cell’s metabolic capacity to assimilate substrates into building blocks. Metabolic genes that determine growth rate may interact synergistically or antagonistically, and can accelerate or slow growth, depending on genetic background and environmental conditions. We evolved a diverse set of Escherichia coli single-gene deletion mutants with a spectrum of growth rates and identified mutations that generally increase growth rate. Despite the metabolic differences between parent strains, mutations that enhanced growth largely mapped to core transcription machinery, including the β and β’ subunits of RNA polymerase (RNAP) and the transcription elongation factor, NusA. The structural segments of RNAP that determine enhanced growth have been previously implicated in antibiotic resistance and in the control of transcription elongation and pausing. We further developed a computational framework to characterize how the transcriptional changes that occur upon acquisition of these mutations affect growth rate across strains. Our experimental and computational results provide evidence for cases in which RNAP mutations shift the competitive balance between active transcription and gene silencing. This study demonstrates that mutations in specific regions of RNAP are a convergent adaptive solution that can enhance the growth rate of cells from distinct metabolic states.

show abstract

Synthetic rescue couples NADPH generation to metabolite overproduction in Saccharomyces cerevisiae

Cited by 16 publications

References 41 publications

Rerouting of NADPH synthetic pathways for increased protopanaxadiol production in Saccharomyces cerevisiae

Rerouting of NADPH synthetic pathways for increased protopanaxadiol production in Saccharomyces cerevisiae

Disorders in NADPH generation via pentose phosphate pathway influence the reproductive potential of the Saccharomyces cerevisiae yeast due to changes in redox status

Experimental evolution of diverse Escherichia coli metabolic mutants identifies genetic loci for convergent adaptation of growth rate

Contact Info

Product

Resources

About