The assimilation of bicarbonate by Neocosmospora vasinfecta

1969 ◽  
Vol 15 (5) ◽  
pp. 389-398 ◽  
Author(s):  
K. Budd

The assimilation of 14C-bicarbonate under controlled conditions was examined in midlog-phase mycelium grown on dextrose as sole carbon source. Sustained assimilation depended on the presence of exogenous nitrogen and carbon sources. When these were provided, assimilation rates of 20–30 μmoles/hour per 100 mg dry weight were maintained for at least 4 hours. After the second hour, almost all of the assimilated bicarbonate-C entered the 80% ethanol-insoluble fraction. Amino acids, especially aspartic and glutamic, were the main destination of assimilated bicarbonate-C; nucleic acids and acids of the tricarboxylic acid cycle accounted for smaller amounts of this carbon. The apparent Km for overall assimilation was 1.4 – 2.2 × 10−4 M with respect to bicarbonate.Assimilation was inhibited by inhibitors of protein synthesis, especially actidione and p-fluorophenylalanine. Evidence was obtained for regulation of assimilation by its end products, and also by the carbon source on which the mycelium was grown. It is concluded that assimilation of bicarbonate or CO2 has an anaplerotic function during protein synthesis in this organism.

1985 ◽  
Vol 31 (8) ◽  
pp. 702-706 ◽  
Author(s):  
Kenneth G. Surowitz ◽  
Robert M. Pfister

The formation of aerial mycelia and spores by Streptomyces alboniger has been observed to be inhibited by glucose supplied in the growth medium as the sole carbon source or supplied in combination with other utilizable carbon sources. Analysis of the metabolism of radiolabelled mannose and sucrose in the presence and absence of glucose demonstrated that glucose functions as the preferred carbon source, inhibiting the uptake and oxidation of the sugars within 15 min of its addition. The inhibition of aerial mycelium formation was shown to result from the excretion of an acidic metabolite, and could be overcome by the addition of a buffering system. The acid metabolite was identified as pyruvic acid by high-performance liquid chromatography and by paper chromatography. Acid was not produced in substantial quantities in dextrin broth or in glucose broth supplemented with 5 mM adenine. Analysis of the pathway of pyruvate overproduction demonstrated that growth on glucose resulted in increased glycolytic activity, relative to the activity of the tricarboxylic acid cycle on this substrate, while growth on dextrin or glucose supplemented with 5 mM adenine resulted in balanced glycolytic and tricarboxylic acid cycle activities.


1986 ◽  
Vol 6 (6) ◽  
pp. 1936-1942
Author(s):  
K S Kim ◽  
M S Rosenkrantz ◽  
L Guarente

The tricarboxylic acid cycle occurs within the mitochondria of the yeast Saccharomyces cerevisiae. A nuclear gene encoding the tricarboxylic acid cycle enzyme citrate synthase has previously been isolated (M. Suissa, K. Suda, and G. Schatz, EMBO J. 3:1773-1781, 1984) and is referred to here as CIT1. We report here the isolation, by an immunological method, of a second nuclear gene encoding citrate synthase (CIT2). Disruption of both genes in the yeast genome was necessary to produce classical citrate synthase-deficient phenotypes: glutamate auxotrophy and poor growth on rich medium containing lactate, a nonfermentable carbon source. Therefore, the citrate synthase produced from either gene was sufficient for these metabolic roles. Transcription of both genes was maximally repressed in medium containing both glucose and glutamate. However, transcription of CIT1 but not of CIT2 was derepressed in medium containing a nonfermentable carbon source. The significance of the presence of two genes encoding citrate synthase in S. cerevisiae is discussed.


Author(s):  
M O Oyewale

The mycelial dry weight and dinitrosalicylic acid (D.N.S.A.) method was used to determine growth and amylase production by Aspergillus flavus grown on different carbon sources. Growth of the fungus was determined at 24 h intervals over a period of six days by the dry mycelial weight methods, while the amylase activity in the culture filtrates of A. flavus was determined by the D.N.S.A method. A total of 45 samples were prepared to determine growth and amylase activity of Aspergillus flavus grown on different carbon sources. The concentration of the various carbon sources ranges between 0.4 to 2% W/V. Duncan’s multiple range test was used to determine the level of significance of the different carbon sources for effective growth and amylase production by Aspergillus flavus. Aspergillus flavus demonstrated the capability to produce significant growth and amylase activities in the medium containing soluble starch, sorghum and cassava peel as sole carbon source. The amount of mycelial dry weight produced from soluble starch, sorghum and cassava peel is significantly higher than those produced from other carbon sources. The data revealed that there is a correlation between growth and amylase production by Aspergillus flavus. The available data from this study showed that soluble starch is the best carbon source for optimum growth and amylase production by A flavus while sorghum and cassava peel are close substitute for optimum growth and amylase production by Aspergillus flavus. Keywords: Growth, amylase activity and Aspergillus flavus


1995 ◽  
Vol 88 (6) ◽  
pp. 687-693 ◽  
Author(s):  
K. Sahlin ◽  
L. Jorfeldt ◽  
K.-G. Henriksson ◽  
S. F. Lewis ◽  
R. G. Haller

1. The importance of the level of tricarboxylic acid cycle intermediates (malate, citrate and fumarate) for energy transduction during exercise has been investigated in six healthy subjects and in two patients with muscle phosphorylase deficiency (McArdle's disease). 2. Healthy subjects cycled for 10 min at low (50 W), moderate [130 ± 6 W (mean ± SEM)] and high (226 ± 12 W) work rates, corresponding to 26, 50 and 80% of their maximal O2 uptake, respectively. Patients with McArdle's disease cycled for 11–13 min at submaximal (40 W) rates, and to fatigue at maximal work rates of 60–90 W. 3. In healthy subjects, phosphocreatine was unchanged during low work rates, but decreased to 79 and 32% of the initial level during moderate and high work rates. In patients with McArdle's disease, phosphocreatine decreased to 82 and 34% of the initial level during submaximal and peak exercise. Muscle lactate increased in healthy subjects during exercise at moderate and high work rates, but remained low in patients with McArdle's disease. 4. In healthy subjects, tricarboxylic acid cycle intermediates were similar at rest and at low work rates (0.48 ± 0.04 mmol/kg dry weight), but increased to 1.6 ± 0.2 mmol/kg dry weight and 4.0 ± 0.3 mmol/kg dry weight at moderate and high work rates. The tricarboxylic acid cycle intermediate level in patients with McArdle's disease was similar to that in healthy subjects at rest, but was markedly reduced during exercise when compared at the same relative intensity. The peak level of tricarboxylic acid cycle intermediates in patients with McArdle's disease was 22% of that in healthy subjects. However, when compared at the same absolute workload, tricarboxylic acid cycle intermediates were similar in patients with McArdle's disease and in healthy subjects. 5. The decrease in glutamate and increase in alanine suggest that the alanine aminotransaminase reaction was the major anaplerotic process in healthy subjects. However, in patients with McArdle's disease (n = 1), muscle alanine remained unchanged and the purine nucleotide cycle may instead be the route of a limited anaplerosis during maximal exercise. The muscle content of glutamate and glutamine (n = 1) was markedly reduced in patients with McArdle's disease. 6. It is concluded that the tricarboxylic acid cycle intermediate level is related to the work rate in healthy subjects, and that the attenuated peak level in patients with McArdle's disease may be a limitation for aerobic energy transduction.


1997 ◽  
Vol 272 (3) ◽  
pp. E476-E484 ◽  
Author(s):  
J. A. Tayek ◽  
J. Katz

Six normal subjects (NL group) and 13 cancer patients (CAI and CAII groups) were fasted overnight and infused with [U-(13)C]glucose (0.016-0.058 mg x min(-1) x kg(-1)). Plasma glucose and lactate were isolated, and mass isotopomer distributions were determined by gas chromatography-mass spectroscopy. Applying equations modified from those previously described [J. A. Tayek and J. Katz. Am. J. Physiol. 270 (Endocrinol. Metab. 33): E709-E717, 1996], we determined glucose production (GP), recycling of glucose carbons, fraction of recycled molecules in blood glucose (Cori cycle), formation of pyruvate from unlabeled carbons, dilution of pyruvate via the tricarboxylic acid cycle and other reactions, and rate of gluconeogenesis. Glucose production was similar in all groups: 2.4 +/- 0.2 mg x min(-1) x kg (-1). The fraction of recycled carbon and of the Cori cycle were elevated in the CAI group vs. the CAII and NL groups: 15 and 33% vs. 7.8 and 19%, respectively (P = 0.01). Gluconeogenesis was 1.9 +/- 0.1, 1.0 +/- 0.1, and 0.83 +/- 0.11 mg x min(-1) x kg(-1) in the CAI, CAII, and NL groups, respectively. In the NL and CAII groups, 20% of GP is via recycling, 20% from unlabeled carbon sources (muscle glycogen, amino acids), and 60% from hepatic glycogenolysis; in the CAI group, 30% is from recycling, 50% from unlabeled carbon, and 20% from glycogen and other sources. Serum cortisol was elevated in the CAI group vs. the CAII group: 11.2 +/- 1.2 vs. 7.7 +/- 1.2 microg/dl (P < 0.05). There was a strong correlation between plasma cortisol and Cori cycle in the NL (r = 0.963) and CAI and CAII groups (r = 0.771). Serum cortisol was directly, and insulin was inversely, correlated with gluconeogenesis in the NL (r2 = 0.967) and CAI and CAII groups (r2 = 0.727). We conclude that whereas the cancer population is heterogeneous with respect to gluconeogenesis, many cancer patients derive their GP predominantly from gluconeogenesis compared with healthy controls, who derive less than one-half of their GP from gluconeogenesis.


2019 ◽  
Vol 93 (13) ◽  
Author(s):  
Anil Pant ◽  
Shuai Cao ◽  
Zhilong Yang

ABSTRACTViruses actively interact with host metabolism because viral replication relies on host cells to provide nutrients and energy. Vaccinia virus (VACV; the prototype poxvirus) prefers glutamine to glucose for efficient replication to the extent that VACV replication is hindered in glutamine-free medium. Remarkably, our data show that VACV replication can be fully rescued from glutamine depletion by asparagine supplementation. By global metabolic profiling, as well as genetic and chemical manipulation of the asparagine supply, we provide evidence demonstrating that the production of asparagine, which exclusively requires glutamine for biosynthesis, accounts for VACV’s preference of glutamine to glucose rather than glutamine’s superiority over glucose in feeding the tricarboxylic acid (TCA) cycle. Furthermore, we show that sufficient asparagine supply is required for efficient VACV protein synthesis. Our study highlights that the asparagine supply, the regulation of which has been evolutionarily tailored in mammalian cells, presents a critical barrier to VACV replication due to a high asparagine content of viral proteins and a rapid demand of viral protein synthesis. The identification of asparagine availability as a critical limiting factor for efficient VACV replication suggests a new direction of antiviral strategy development.IMPORTANCEViruses rely on their infected host cells to provide nutrients and energy for replication. Vaccinia virus, the prototypic member of the poxviruses, which comprise many significant human and animal pathogens, prefers glutamine to glucose for efficient replication. Here, we show that the preference is not because glutamine is superior to glucose as the carbon source to fuel the tricarboxylic acid cycle for vaccinia virus replication. Rather interestingly, the preference is because the asparagine supply for efficient viral protein synthesis becomes limited in the absence of glutamine, which is necessary for asparagine biosynthesis. We provide further genetic and chemical evidence to demonstrate that asparagine availability plays a critical role in efficient vaccinia virus replication. This discovery identifies a weakness of vaccinia virus and suggests a possible direction to intervene in poxvirus infection.


2006 ◽  
Vol 74 (2) ◽  
pp. 1130-1140 ◽  
Author(s):  
Merlin Tchawa Yimga ◽  
Mary P. Leatham ◽  
James H. Allen ◽  
David C. Laux ◽  
Tyrrell Conway ◽  
...  

ABSTRACT In Salmonella enterica serovar Typhimurium, the Cra protein (catabolite repressor/activator) regulates utilization of gluconeogenic carbon sources by activating transcription of genes in the gluconeogenic pathway, the glyoxylate bypass, the tricarboxylic acid (TCA) cycle, and electron transport and repressing genes encoding glycolytic enzymes. A serovar Typhimurium SR-11 Δcra mutant was recently reported to be avirulent in BALB/c mice via the peroral route, suggesting that gluconeogenesis may be required for virulence. In the present study, specific SR-11 genes in the gluconeogenic pathway were deleted (fbp, glpX, ppsA, and pckA), and the mutants were tested for virulence in BALB/c mice. The data show that SR-11 does not require gluconeogenesis to retain full virulence and suggest that as yet unidentified sugars are utilized by SR-11 for growth during infection of BALB/c mice. The data also suggest that the TCA cycle operates as a full cycle, i.e., a sucCD mutant, which prevents the conversion of succinyl coenzyme A to succinate, and an ΔsdhCDA mutant, which blocks the conversion of succinate to fumarate, were both attenuated, whereas both an SR-11 ΔaspA mutant and an SR-11 ΔfrdABC mutant, deficient in the ability to run the reductive branch of the TCA cycle, were fully virulent. Moreover, although it appears that SR-11 replenishes TCA cycle intermediates from substrates present in mouse tissues, fatty acid degradation and the glyoxylate bypass are not required, since an SR-11 ΔfadD mutant and an SR-11 ΔaceA mutant were both fully virulent.


2002 ◽  
Vol 68 (8) ◽  
pp. 3848-3854 ◽  
Author(s):  
Ilana S. Aldor ◽  
Seon-Won Kim ◽  
Kristala L. Jones Prather ◽  
Jay D. Keasling

ABSTRACT A pathway was metabolically engineered to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a biodegradable thermoplastic with proven commercial applications, from a single, unrelated carbon source. An expression system was developed in which a prpC strain of Salmonella enterica serovar Typhimurium, with a mutation in the ability to metabolize propionyl coenzyme A (propionyl-CoA), served as the host for a plasmid harboring the Acinetobacter polyhydroxyalkanoate synthesis operon (phaBCA) and a second plasmid with the Escherichia coli sbm and ygfG genes under an independent promoter. The sbm and ygfG genes encode a novel (2R)-methylmalonyl-CoA mutase and a (2R)-methylmalonyl-CoA decarboxylase, respectively, which convert succinyl-CoA, derived from the tricarboxylic acid cycle, to propionyl-CoA, an essential precursor of 3-hydroxyvalerate (HV). The S. enterica system accumulated PHBV with significant HV incorporation when the organism was grown aerobically with glycerol as the sole carbon source. It was possible to vary the average HV fraction in the copolymer by adjusting the arabinose or cyanocobalamin (precursor of coenzyme B12) concentration in the medium.


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