A constraint-based model analysis of the metabolic consequences of increased NADPH oxidation in Saccharomyces cerevisiae

被引:60
作者
Celton, Magalie [1 ,2 ,3 ,4 ]
Goelzer, Anne [4 ]
Camarasa, Carole [1 ,2 ,3 ]
Fromion, Vincent [4 ]
Dequin, Sylvie [1 ,2 ,3 ]
机构
[1] INRA, UMR1083, F-34060 Montpellier, France
[2] SupAgro, UMR1083, Sci Oenol, F-34060 Montpellier, France
[3] Univ Montpellier I, UMR1083, F-34060 Montpellier, France
[4] INRA, Math Informat & Genome UR1077, F-78350 Jouy En Josas, France
关键词
NADPH oxidation; Constraint-based modeling; Redox cycles; PENTOSE-PHOSPHATE PATHWAY; ALDO-KETO REDUCTASE; YEAST-CELLS; 2,3-BUTANEDIOL DEHYDROGENASE; FLUX DISTRIBUTIONS; GLYCEROL FORMATION; PRODUCT FORMATION; GENE-PRODUCT; GROWTH; GLUCOSE;
D O I
10.1016/j.ymben.2012.03.008
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Controlling the amounts of redox cofactors to manipulate metabolic fluxes is emerging as a useful approach to optimizing byproduct yields in yeast biotechnological processes. Redox cofactors are extensively interconnected metabolites, so predicting metabolite patterns is challenging and requires in-depth knowledge of how the metabolic network responds to a redox perturbation. Our aim was to analyze comprehensively the metabolic consequences of increased cytosolic NADPH oxidation during yeast fermentation. Using a genetic device based on the overexpression of a modified 2,3-butanediol dehydrogenase catalyzing the NADPH-dependent reduction of acetoin into 2,3-butanediol, we increased the NADPH demand to between 8 and 40-fold the anabolic demand. We developed (i) a dedicated constraint-based model of yeast fermentation and (ii) a constraint-based modeling method based on the dynamical analysis of mass distribution to quantify the in vivo contribution of pathways producing NADPH to the maintenance of redox homeostasis. We report that yeast responds to NADPH oxidation through a gradual increase in the flux through the PP and acetate pathways, providing 80% and 20% of the NADPH demand, respectively. However, for the highest NADPH demand, the model reveals a saturation of the PP pathway and predicts an exchange between NADH and NADPH in the cytosol that may be mediated by the glycerol-DHA futile cycle. We also reveal the contribution of mitochondrial shuttles, resulting in a net production of NADH in the cytosol, to fine-tune the NADH/NAD(+) balance. This systems level study helps elucidate the physiological adaptation of yeast to NADPH perturbation. Our findings emphasize the robustness of yeast to alterations in NADPH metabolism and highlight the role of the glycerol-DHA cycle as a redox valve, providing additional NADPH from NADH under conditions of very high demand. (C) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码:366 / 379
页数:14
相关论文
共 78 条
[1]   The compositions of wax esters, triacylglycerols and phospholipids in Arctic and Antarctic copepods: Evidence of energetic adaptations [J].
Albers, CS ;
Kattner, G ;
Hagen, W .
MARINE CHEMISTRY, 1996, 55 (3-4) :347-358
[2]  
Ambroset C., 2011, G3, DOI http://dx.doi.org/10.1534/g3.111.000422
[3]  
Anderlund M, 1999, APPL ENVIRON MICROB, V65, P2333
[4]  
[Anonymous], 2013, Introductory lectures on convex optimization: A basic course
[5]  
[Anonymous], 1989, Molecular Cloning: A Laboratory
[6]   The mitochondrial alcohol dehydrogenase adh3p is involved in a redox shuttle in Saccharomyces cerevisiae [J].
Bakker, BM ;
Bro, C ;
Kötter, P ;
Luttik, MAH ;
van Dijken, JP ;
Pronk, JT .
JOURNAL OF BACTERIOLOGY, 2000, 182 (17) :4730-4737
[7]  
Bakker BM, 2001, FEMS MICROBIOL REV, V25, P15, DOI 10.1016/S0168-6445(00)00039-5
[8]   Large-scale 13C-flux analysis reveals mechanistic principles of metabolic network robustness to null mutations in yeast -: art. no. R49 [J].
Blank, LM ;
Kuepfer, L ;
Sauer, U .
GENOME BIOLOGY, 2005, 6 (06)
[9]  
BLIGH EG, 1959, CAN J BIOCHEM PHYS, V37, P911
[10]   PHYSIOLOGY OF OSMOTOLERANCE IN FUNGI [J].
BLOMBERG, A ;
ADLER, L .
ADVANCES IN MICROBIAL PHYSIOLOGY, 1992, 33 :145-212