Time-course of enzyme adaptation. I. Effects of substituting dietary glucose and fructose at constant concentrations of dietary protein

1968 ◽  
Vol 46 (12) ◽  
pp. 1459-1470 ◽  
Author(s):  
B. Szepesi ◽  
R. A. Freedland

The effect of dietary fructose on liver, kidney, and adrenal enzymes was studied in male Sprague–Dawley rats. Dietary fructose increased relative liver and kidney sizes, liver glycogen, and liver protein. The percentage increases in relative liver and kidney sizes were independent of dietary protein, but relative liver sizes were smaller in the absence of protein.The activities of all the liver enzymes studied, except glucokinase, were increased by a 65% fructose diet containing 25% casein. The rates of increases differed between enzymes; the activities of L-α-glycerophosphate dehydrogenase and phosphoenolpyruvate carboxykinase (PEP-carboxykinase) reached almost maximum in 1 day, glucose 6-phosphatase, 6-phosphogluconate dehydrogenase, and pyruvate kinase activities reached maximum in about 2 days, and glucose 6-phosphate dehydrogenase, malic enzyme, dihydroxyacetone kinase, phosphofructose kinase, and phosphohexose isomerase required at least 3 days to reach maximum activity after the dietary change. The increases in the activities of liver fructose 1,6-diphosphatase, glucose 6-phosphate dehydrogenase, pyruvate kinase, and phosphohexose isomerase did not occur in the absence of dietary protein. The activities of liver phosphofructokinase and malic enzyme were increased equally well whether the fructose diet contained protein or not, while the increases in the activities of other enzymes were less in the absence of dietary protein.The half-life of liver malic enzyme was estimated as 3 days in the glucose to fructose dietary change and 1 day in the fructose to glucose dietary change. Since malic enzyme activity was increased by fructose feeding about threefold, the data suggest that under the conditions of the experiment fructose increased malic enzyme activity by decreasing the rate of breakdown of the enzyme.In general, kidney enzymes were affected by fructose to a lesser extent than the corresponding enzymes in liver. This was particularly significant in the case of kidney glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and malic enzyme, the activities of which were only slightly increased in kidney. The activities of these three enzymes in the adrenal glands were not increased by fructose feeding.

1967 ◽  
Vol 105 (2) ◽  
pp. 717-722 ◽  
Author(s):  
C B Taylor ◽  
E. Bailey ◽  
W Bartley

1. Changes in the activities of ATP citrate lyase, ‘malic’ enzyme, glucose 6-phosphate dehydrogenase, pyruvate kinase and fructose 1,6-diphosphatase, and in the ability to incorporate [1−14C]acetate into lipid have been measured in the livers of developing rats between late foetal life and maturity. 2. In male rats the activities of those systems directly or indirectly concerned in lipogenesis (acetate incorporation into lipid, ATP citrate lyase and glucose 6-phosphate dehydrogenase) fall after birth and are maintained at a low value until weaning. After weaning these activities rise to a maximum between 30 and 40 days and then decline, reaching adult values at about 60 days. ‘Malic’ enzyme activity follows a similar course, except that none could be detected in the foetal liver. Pyruvate kinase activity is lower in foetal than in adult livers and rises to slightly higher than the adult value in the post-weaning period. Fructose 1,6-diphosphatase activity rises from a very low foetal value to reach a maximum at about 10 days but falls rapidly after weaning to reach adult values at about 30 days. 3. Weaning rats on to a high-fat diet caused the low activities of acetate incorporation, ATP citrate lyase, glucose 6-phosphate dehydrogenase and pyruvate kinase, characteristic of the suckling period, to persist. ‘Malic’ enzyme and fructose 1,6-diphosphatase activities were not altered appreciably. 4. No differences could be detected in hepatic enzyme activities between males and females up to 35 days, but after this time female rats gave higher values for acetate incorporation, glucose 6-phosphate dehydrogenase activity and ‘malic’ enzyme activity. 5. The results are discussed in relation to changes in alimentation and hormonal influences.


1998 ◽  
Vol 180 (11) ◽  
pp. 2875-2882 ◽  
Author(s):  
Eckhard Boles ◽  
Patricia de Jong-Gubbels ◽  
Jack T. Pronk

ABSTRACT Pyruvate, a precursor for several amino acids, can be synthesized from phosphoenolpyruvate by pyruvate kinase. Nevertheless, pyk1 pyk2 mutants of Saccharomyces cerevisiae devoid of pyruvate kinase activity grew normally on ethanol in defined media, indicating the presence of an alternative route for pyruvate synthesis. A candidate for this role is malic enzyme, which catalyzes the oxidative decarboxylation of malate to pyruvate. Disruption of open reading frame YKL029c, which is homologous to malic enzyme genes from other organisms, abolished malic enzyme activity in extracts of glucose-grown cells. Conversely, overexpression ofYKL029c/MAE1 from the MET25 promoter resulted in an up to 33-fold increase of malic enzyme activity. Growth studies with mutants demonstrated that presence of either Pyk1p or Mae1p is required for growth on ethanol. Mutants lacking both enzymes could be rescued by addition of alanine or pyruvate to ethanol cultures. Disruption of MAE1 alone did not result in a clear phenotype. Regulation of MAE1 was studied by determining enzyme activities and MAE1 mRNA levels in wild-type cultures and by measuring β-galactosidase activities in a strain carrying a MAE1::lacZ fusion. Both in shake flask cultures and in carbon-limited chemostat cultures,MAE1 was constitutively expressed. A three- to fourfold induction was observed during anaerobic growth on glucose. Subcellular fractionation experiments indicated that malic enzyme in S. cerevisiae is a mitochondrial enzyme. Its regulation and localization suggest a role in the provision of intramitochondrial NADPH or pyruvate under anaerobic growth conditions. However, since null mutants could still grow anaerobically, this function is apparently not essential.


2009 ◽  
Vol 78 (1) ◽  
pp. 179-185 ◽  
Author(s):  
Olcay Hisar ◽  
Adem Yavuz Sönmez ◽  
Şükrü Beydemir ◽  
Şükriye Aras Hisar ◽  
Telat Yanik ◽  
...  

The effects of cadmium (Cd) on the enzymatic activities of glucose 6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD) were investigated in the gill, liver and kidney tissues of rainbow trout (Oncorhynchus mykiss). Three test groups of fish were subjected to increasing concentrations (1, 3 and 5 mg/l) of cadmium (Cd) in vivo, respectively. The G6PD and 6PGD activities in the gill, liver, and kidney tissues of each group of fish were measured on days 1, 3, 5 and 7. G6PD and 6PGD enzyme activities, measured in gill, liver and kidney homogenates, were stimulated by various concentrations (1, 3, and 5 mg/l) of cadmium. Although the dose-response pattern of G6PD enzyme activities in liver and kidney tissue was very similar, that in gill was different from both other tissues. The enzyme activity of G6PD enzyme was significantly stimulated after three days (Day 3) in liver and kidney tissues at a dose of 1 mg/l Cd (p < 0.05), whereas it was stimulated on the first day of experiment (Day 1) in gill, liver and kidney tissues at doses of 3 and 5 mg/l Cd (p < 0.05). However, the activity of 6PGD was stimulated after three days (Day 3) in the liver at a dose of 1 mg/l Cd (p < 0.05) and on the first day in gill, liver and kidney tissues at doses of 3 and 5 mg/l Cd (p < 0.05). The stimulation effect of the 5 mg/l dose of Cd on G6PD and 6PGD enzyme activities was significantly diminished after seven days (Day 7) in all tissues (p < 0.05). In contrast to the dose-response pattern at the dose of 5 mg/l Cd, G6PD and 6PGD enzyme activities were stimulated significantly (p < 0.05) in liver and kidney tissues at the doses of 3 and 1 mg/l Cd. The stimulation effect of cadmium on the three tissues studied was also calculated; for both of the enzymes (G6PD and 6PGD), the enzyme activity levels were stimulated by approximately 60% and 38% in gills, 68% and 44% in liver, and 67% and 41% in kidneys, respectively, over the base-line enzyme activity of the control groups during the sevenday experimental period. These findings indicate that tissue G6PD and 6PGD enzymes function to protect against cadmium toxicity.


Author(s):  
S.M. Geyer ◽  
C.L. Mendenhall ◽  
J.T. Hung ◽  
E.L. Cardell ◽  
R.L. Drake ◽  
...  

Thirty-three mature male Holtzman rats were randomly placed in 3 treatment groups: Controls (C); Ethanolics (E); and Wine drinkers (W). The animals were fed synthetic diets (Lieber type) with ethanol or wine substituted isocalorically for carbohydrates in the diet of E and W groups, respectively. W received a volume of wine which provided the same gram quantity of alcohol consumed by E. The animals were sacrificed by decapitation after 6 weeks and the livers processed for quantitative triglycerides (T3), proteins, malic enzyme activity (MEA), light microscopy (LM) and electron microscopy (EM). Morphometric analysis of randomly selected LM and EM micrographs was performed to determine organellar changes in centrilobular (CV) and periportal (PV) regions of the liver. This analysis (Table 1) showed that hepatocytes from E were larger than those in C and W groups. Smooth endoplasmic reticulum decreased in E and increased in W compared to C values.


Nature ◽  
1956 ◽  
Vol 177 (4514) ◽  
pp. 842-843 ◽  
Author(s):  
S. E. LEWIS ◽  
G. M. PRICE

Development ◽  
1984 ◽  
Vol 80 (1) ◽  
pp. 251-288
Author(s):  
R. L. Gardner

Conditions were found for staining whole mid-gestation capsular parietal endoderms and visceral yolk sacs for malic enzyme activity that gave excellent discrimination between wildtype (Mod-1+/Mod-1+) cells and mutant (Mod-ln/Mod-1n) cells that lack the cytoplasmic form of the enzyme. Reciprocal blastocyst injection experiments were undertaken in which single primitive endoderm cells of one genotype were transplanted into embryos of the other genotype. In addition, Mod-1+/Mod-1+ early inner cell mass (ICM) cells were injected into Mod-1n/Mod-1n blastocysts, either in groups of two or three singletons or as daughter cell pairs. A substantial proportion of the resulting conceptuses showed mosaic histochemical staining in the parietal endoderm, visceral yolk sac, or in both these membranes. Stained cells were invariably intimately intermixed with unstained cells in the mosaic parietal endoderms. In contrast, one or both of two distinct patterns of staining could be discerned in mosaic visceral yolk sacs. The first, a conspicuously ‘coherent’ pattern, was found to be due to endodermal chimaerism; the second, a more diffuse pattern, was attributable to chimaerism in the mesodermal layer of this membrane. The overall distribution of cells with donor staining characteristics resulting from primitive endoderm versus early ICM cell injections was consistent with findings in earlier experiments in which allozymes of glucosephosphate isomerase were used as markers. The conspicuous lack of phenotypically intermediate cells in predominantly stained areas of mosaic membranes suggested that the histochemical difference between Mod-1+/Mod-1+ and Mod-1n/Mod-ln genotypes was cell-autonomous. This conclusion was strengthened by the results of staining mixed in vitro cultures of parietal endoderm in which presence or absence of phagocytosed melanin granules was used as an independent means of distinguishing wild type from null cells. By substituting tetranitro blue tetrazolium for nitro blue tetrazolium in the incubation medium, satisfactory differential staining was obtained for both the extraembryonic endoderm and other tissues of earlier postimplantation wild type versus null embryos. Finally, absence of cytoplasmic malic enzyme activity does not appear to have a significant effect on the viability or behaviour of mutant cells.


2006 ◽  
Vol 188 (13) ◽  
pp. 4727-4736 ◽  
Author(s):  
Guillaume Lerondel ◽  
Thierry Doan ◽  
Nicola Zamboni ◽  
Uwe Sauer ◽  
Stéphane Aymerich

ABSTRACT The Bacillus subtilis genome contains several sets of paralogs. An extreme case is the four putative malic enzyme genes maeA, malS, ytsJ, and mleA. maeA was demonstrated to encode malic enzyme activity, to be inducible by malate, but also to be dispensable for growth on malate. We report systematic experiments to test whether these four genes ensure backup or cover different functions. Analysis of single- and multiple-mutant strains demonstrated that ytsJ has a major physiological role in malate utilization for which none of the other three genes could compensate. In contrast, maeA, malS, and mleA had distinct roles in malate utilization for which they could compensate one another. The four proteins exhibited malic enzyme activity; MalS, MleA, and MaeA exhibited 4- to 90-fold higher activities with NAD+ than with NADP+. YtsJ activity, in contrast, was 70-fold higher with NADP+ than with NAD+, with Km values of 0.055 and 2.8 mM, respectively. lacZ fusions revealed strong transcription of ytsJ, twofold higher in malate than in glucose medium, but weak transcription of malS and mleA. In contrast, mleA was strongly transcribed in complex medium. Metabolic flux analysis confirmed the major role of YtsJ in malate-to-pyruvate interconversion. While overexpression of the NADP-dependent Escherichia coli malic enzyme MaeB did not suppress the growth defect of a ytsJ mutant on malate, overexpression of the transhydrogenase UdhA from E. coli partially suppressed it. These results suggest an additional physiological role of YtsJ beyond that of malate-to-pyruvate conversion.


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