RESUMO
Cultures of Shewanella putrefaciens grown in medium containing 10mM 1,4-diamino-2-butanone (DBO) as an inhibitor of ornithine decarboxylase and 10mM 1,5-diaminopentane (cadaverine) showed the simultaneous biosynthesis of the macrocyclic dihydroxamic acids: putrebactin (pbH2), avaroferrin (avH2) and bisucaberin (bsH2). The level of DBO did not completely repress the production of endogenous 1,4-diaminobutane (putrescine) as the native diamine substrate of pbH2. The relative concentration of pbH2:avH2:bsH2 was 1:2:1, which correlated with the substrate selection of putrescine:cadaverine in a ratio of 1:1. The macrocycles were characterised using LC-MS as free ligands and as 1:1 complexes with Fe(III) of the form [Fe(pb)]+, [Fe(av)]+ or [Fe(bs)]+, with labile ancillary ligands in six-coordinate complexes displaced during ESI-MS acquisition; or with Mo(VI) of the form [Mo(O)2(pb)], [Mo(O)2(av)] or [Mo(O)2(bs)]. Chromium(V) complexes of the form [CrO(pb)]+ were detected from solutions of Cr(VI) and pbH2 in DMF using X-band EPR spectroscopy. Supplementation of S. putrefaciens medium with DBO and 1,3-diaminopropane, 1,6-diaminohexane or 1,4-diamino-2(Z)-butene (Z-DBE) resulted only in the biosynthesis of pbH2. The work has identified a native system for the simultaneous biosynthesis of a suite of three macrocyclic dihydroxamic acid siderophores and highlights both the utility of precursor-directed biosynthesis for expanding the structural diversity of siderophores, and the breadth of their coordination chemistry.
Assuntos
Cromo/química , Ferro/química , Molibdênio/química , Peptídeos Cíclicos/biossíntese , Putrescina/análogos & derivados , Shewanella putrefaciens/metabolismo , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cadaverina/metabolismo , Complexos de Coordenação/química , Diaminas/farmacologia , Espectroscopia de Ressonância de Spin Eletrônica , Expressão Gênica , Ácidos Hidroxâmicos/antagonistas & inibidores , Ornitina Descarboxilase/genética , Ornitina Descarboxilase/metabolismo , Inibidores da Ornitina Descarboxilase/farmacologia , Peptídeos Cíclicos/antagonistas & inibidores , Putrescina/antagonistas & inibidores , Putrescina/biossíntese , Putrescina/farmacologia , Shewanella putrefaciens/efeitos dos fármacos , Shewanella putrefaciens/genética , Succinatos/antagonistas & inibidoresRESUMO
The impact of carvacrol at different levels (0.1%, 0.5%, and 1%) on ammonia (AMN) and biogenic amines (BAs) production by 8 common foodborne pathogens (FBPs) (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Pseudomonas aeruginosa, Listeria monocytogenes, Aeromonas hydrophila, and Salmonella Paratyphi A) was studied using a rapid high-performance liquid chromatography method. Significant differences among bacteria (P < 0.05) in AMN and BA production were observed using a tyrosine decarboxylase broth. Tyramine, dopamine, agmatine, spermine, and putrescine were the main amines produced by the bacteria. Tyramine production by P. aeruginosa was the highest (967 mg/L), whereas K. pneumoniae was the poorest tyramine producer (6.42 mg/L). AMN and BA production varied significantly depending on carvacrol levels and the specific bacterial strains. Tyramine production for all bacterial strains was significantly suppressed by addition of carvacrol at levels of 0.5% and 1%, but not 0.1%. Consequently, the effect of carvacrol on BA and AMN formation by FBP was dependent on bacterial strain as well as carvacrol level.
Assuntos
Amônia/metabolismo , Bactérias/efeitos dos fármacos , Aminas Biogênicas/biossíntese , Doenças Transmitidas por Alimentos/microbiologia , Monoterpenos/farmacologia , Extratos Vegetais/farmacologia , Bactérias/metabolismo , Cimenos , Humanos , Putrescina/biossíntese , Tiramina/biossínteseRESUMO
Freeze-dried black raspberries (BRBs) have demonstrated chemopreventive effects in a dietary intervention trial with human colorectal cancer patients. The aim of this study was to investigate BRB-caused metabolite changes using the Apc(Min/+) mouse as a model of human colorectal cancer. Wild-type (WT) mice were fed control diet, and Apc(Min/+) mice were fed either control diet or control diet supplemented with 5% BRBs for 8 weeks. Colonic and intestinal polyp size and number were measured. A non-targeted metabolomic analysis was conducted on colonic mucosa, liver and fecal specimens. Eight weeks of BRB treatment significantly decreased intestinal and colonic polyp number and size in Apc(Min/+) mice. The apc gene mutation significantly changed 52 metabolites in colonic mucosa associated with increased amino acid and decreased lipid metabolites, as well as 39 liver and 8 fecal metabolites. BRBs significantly reversed 23 apc-regulated metabolites, including 13 colonic mucosa, 8 liver and 2 fecal metabolites that were involved in amino acid, glutathione, lipid and nucleotide metabolism. Of these, changes in eight metabolites were linearly correlated with decreased colonic polyp number and size in BRB-treated Apc(Min/+) mice. Elevated levels of putrescine and linolenate in Apc(Min/+) mice were significantly decreased by BRBs. Ornithine decarboxylase expression, the key enzyme in putrescine generation, was fully suppressed by BRBs. These results suggest that BRBs produced beneficial effects against colonic adenoma development in Apc(Min/+) mice and modulated multiple metabolic pathways. The metabolite changes produced by BRBs might potentially reflect the BRB-mediated chemopreventive effects in colorectal cancer patients.
Assuntos
Adenoma/dietoterapia , Proteína da Polipose Adenomatosa do Colo/genética , Neoplasias Colorretais/dietoterapia , Frutas , Rubus , Adenoma/metabolismo , Adenoma/patologia , Animais , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Modelos Animais de Doenças , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Mucosa Intestinal/efeitos dos fármacos , Camundongos , Camundongos Transgênicos , Putrescina/biossíntese , Ácido alfa-Linolênico/biossínteseRESUMO
Corynebacterium glutamicum shows a great potential for the production of the polyamide monomer putrescine (1,4-diaminobutane). Previously, we constructed the putrescine-producing strain PUT1 by deletion of argF, the gene for ornithine transcarbamoylase (OTC), and argR, encoding the L-arginine repressor, combined with heterologous expression of the Escherichia coli gene for L-ornithine decarboxylase SpeC. As a consequence of argF deletion, this strain requires supplementation of L-arginine and shows growth-decoupled putrescine production. To avoid costly supplementation with L-arginine and the strong feedback inhibition of the key enzyme N-acetylglutamate kinase (ArgB) by L-arginine, a plasmid addiction system for low-level argF expression was developed. By fine-tuning argF expression through modifications of the promoter, the translational start codon and/or the ribosome binding site, high productivity and titer could be obtained. OTC activity varied almost thousandfold between 960 and 1 mU mg⻹ resulting in putrescine yields on glucose from less than 0.001 up to 0.26 g g⻹, the highest yield in bacteria reported to date. The most promising strain, designated PUT21, was characterized comprehensively. PUT21 strain grew with a rate of 0.19 h⻹ in mineral salt medium without the need for L-arginine supplementation and produced putrescine with a yield of 0.16 g g⻹ glucose at a volumetric productivity of 0.57 g L⻹ h⻹ and a specific productivity of 0.042 g g⻹ h⻹. The carbon balance suggested that no major unidentified by-product was produced. Compared to the first-generation strain PUT1, the putrescine yield observed with PUT21 was increased by 60%. In fed-batch cultivation with C. glutamicum PUT21, a putrescine titer of 19 g L⻹ at a volumetric productivity of 0.55 g L⻹ h⻹ and a yield of 0.16 g g⻹ glucose could be achieved. Moreover, while plasmid segregation of the initial strain required antibiotic selection, plasmid segregation in C. glutamicum PUT21 was fully stable for more than 60 generations without antibiotic selection even in the presence of L-arginine. The ornithine decarboxylase gene speC was expressed from this argF addiction plasmid ensuring stable putrescine production by the engineered C. glutamicum strain.
Assuntos
Corynebacterium glutamicum/enzimologia , Engenharia Genética/métodos , Ornitina Carbamoiltransferase/metabolismo , Plasmídeos/genética , Putrescina/biossíntese , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biotecnologia , Corynebacterium glutamicum/genética , Corynebacterium glutamicum/metabolismo , Meios de Cultura , Escherichia coli/enzimologia , Escherichia coli/genética , Escherichia coli/metabolismo , Fermentação , Genes Bacterianos , Ornitina Carbamoiltransferase/genéticaRESUMO
AIMS: Lactobacillus brevis IOEB 9809 is able to produce both tyramine and putrescine via tyrosine decarboxylase and agmatine deiminase enzymes, respectively, when cultured on synthetic media. The aims of this study were to assess the expression of L. brevis IOEB 9809 tdc and aguA1 genes, during wine fermentation and to evaluate the effect of substrate availability and pH on tdc and aguA1 expression, as well as on biogenic amine production and L. brevis viability. METHODS AND RESULTS: The relative expression of L. brevis IOEB 9809 tdc and aguA1 genes was analysed in wine by quantitative real-time RT-PCR (qRT-PCR) during a period of incubation of 30 days. Cell viability, pH values, putrescine and tyramine concentration were monitored throughout the experiments. CONCLUSIONS: The wine trials indicated that L. brevis IOEB 9809 is able to produce both tyramine and putrescine during wine fermentation. Increased cell viability was also observed in wine supplemented with tyrosine or agmatine. qRT-PCR analysis suggests a strong influence of substrate availability on the expression of genes coding for tyrosine decarboxylase and agmatine deiminase in L. brevis IOEB 9809. Less evident is the relationship between putrescine and tyramine production and tolerance to wine pH. SIGNIFICANCE AND IMPACT OF STUDY: To our knowledge, this study represents the first assessment of relative expression of L. brevis IOEB 9809 genes involved in biogenic amine production in wine. Furthermore, an effect of biogenic amine production on viability of L. brevis during wine fermentation was established.
Assuntos
Hidrolases/metabolismo , Levilactobacillus brevis/enzimologia , Tirosina Descarboxilase/metabolismo , Vinho/microbiologia , Agmatina/metabolismo , Aminas Biogênicas/análise , Aminas Biogênicas/metabolismo , Fermentação , Humanos , Hidrolases/genética , Levilactobacillus brevis/genética , Putrescina/biossíntese , Putrescina/metabolismo , Tiramina/biossíntese , Tiramina/metabolismo , Tirosina/metabolismo , Tirosina Descarboxilase/genéticaRESUMO
This article evaluates changes in biogenic amines and how these relate to microbiological growth in chilled, fresh restructured beef steaks containing transglutaminase as a cold binding agent and different amounts of walnut. Added walnut and chilling favored higher total and lactic acid bacteria counts during storage, whereas Enterobacteriaceae were not affected. The highest initial biogenic amine concentrations were identified as spermidine, spermine, and tyramine. Both added walnut and cold storage generally favored the formation of amines (tyramine, histamine, putrescine, and cadaverine), which was more obviously apparent by the end of the storage period. Agmatine, on the other hand, was not generally affected by the walnut.
Assuntos
Aminas Biogênicas/análise , Manipulação de Alimentos/métodos , Juglans , Produtos da Carne/microbiologia , Extratos Vegetais/farmacologia , Animais , Aminas Biogênicas/biossíntese , Cadaverina/análise , Cadaverina/biossíntese , Bovinos , Temperatura Baixa , Relação Dose-Resposta a Droga , Histamina/análise , Histamina/biossíntese , Putrescina/análise , Putrescina/biossíntese , Espermidina/análise , Espermidina/biossíntese , Espermina/análise , Espermina/biossíntese , Fatores de Tempo , Transglutaminases/metabolismoRESUMO
We posed the question of whether steady-state levels of the higher polyamines spermidine and spermine in plants can be influenced by overexpression of a heterologous cDNA involved in the later steps of the pathway, in the absence of any further manipulation of the two synthases that are also involved in their biosynthesis. Transgenic rice (Oryza sativa) plants engineered with the heterologous Datura stramonium S-adenosylmethionine decarboxylase (samdc) cDNA exhibited accumulation of the transgene steady-state mRNA. Transgene expression did not affect expression of the orthologous samdc gene. Significant increases in SAMDC activity translated to a direct increase in the level of spermidine, but not spermine, in leaves. Seeds recovered from a number of plants exhibited significant increases in spermidine and spermine levels. We demonstrate that overexpression of the D. stramonium samdc cDNA in transgenic rice is sufficient for accumulation of spermidine in leaves and spermidine and spermine in seeds. These findings suggest that increases in enzyme activity in one of the two components of the later parts of the pathway leading to the higher polyamines is sufficient to alter their levels mostly in seeds and, to some extent, in vegetative tissue such as leaves. Implications of our results on the design of rational approaches for the modulation of the polyamine pathway in plants are discussed in the general framework of metabolic pathway engineering.
Assuntos
Adenosilmetionina Descarboxilase/genética , Oryza/genética , Poliaminas/metabolismo , Adenosilmetionina Descarboxilase/metabolismo , Amina Oxidase (contendo Cobre)/metabolismo , Carboxiliases/metabolismo , DNA Complementar/genética , Datura/enzimologia , Datura/genética , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Teste de Complementação Genética , Ornitina Descarboxilase/metabolismo , Oryza/enzimologia , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/genética , Putrescina/biossíntese , Sementes/metabolismo , Espermidina/biossíntese , Espermina/biossíntese , Poliamina OxidaseRESUMO
Pectin is a soluble indigestible polysaccharide that stimulates cecal polyamine formation in rats. Bacteroides and fusobacteria, two numerically dominant bacterial population groups in the large intestine, were found to synthesize in vitro high amounts of spermidine and putrescine. The purpose of this study was to elucidate the effect of pectin on the polyamine production by defined bacterial species in vivo. Germfree male Wistar rats (n = 18) were randomly assigned to one of three treatments: (i) monoassociation with Bacteroides thetaiotaomicron + fiber-free diet; (ii) diassociation with B. thetaiotaomicron + Fusobacterium varium + fiber-free diet or (iii) diassociation with B. thetaiotaomicron + F. varium + fiber-free diet + 10% pectin. The cecal contents of monoassociated rats fed fiber-free diet contained large amounts (1.51+/-0.21 micromol/dry total cecum content) of spermidine which was the major polyamine. The cecum of diassociated rats fed the fiber-free diet contained even higher concentrations of spermidine (2.53+/-0.21 micromol/dry total cecum content) and also putrescine, which was now the dominant polyamine (putrescine 0.32+/-0.28 vs. 3.01+/-0.28 micromol/dry total cecum content; monoassociation vs. diassociation). Pectin consumption by diassociated rats led to an additional increase in the cecal concentrations of all polyamines: putrescine, spermidine and spermine were 40, 37 and 100%, respectively, higher in the diassociated rats consuming the pectin diet than in those consuming the pectin-free diet. Since the microbial counts in the cecum did not differ in the diassociated treatment groups, the elevated concentrations of polyamines observed in the pectin group must have been due to stimulated bacterial polyamine synthesis. The decline of individual polyamines from cecum to feces detected at the end of the study in all treatment groups and the high microbial counts in the cecum and in feces suggest that bacterial polyamines are absorbed in cecum and colon. Pectin stimulates intestinal microbes to synthesize large amounts of polyamines which may be utilized by the host.
Assuntos
Bacteroides/metabolismo , Ceco/microbiologia , Dieta , Fibras na Dieta/administração & dosagem , Fusobacterium/metabolismo , Pectinas/administração & dosagem , Putrescina/biossíntese , Espermidina/biossíntese , Análise de Variância , Animais , Ceco/efeitos dos fármacos , Ingestão de Energia , Fezes/microbiologia , Humanos , Concentração de Íons de Hidrogênio , Masculino , Pectinas/farmacologia , Ratos , Ratos Wistar , Aumento de PesoRESUMO
The effects of two highly fermentable dietary fibers (guar gum and pectin) on the type and concentrations of cecal polyamines as affected by the intestinal microflora were studied in groups of germ-free (n = 10/group) and conventional rats (n = 6/group). Both germ-free and conventional rats were randomly assigned to one of three treatments as follows: 1) fiber-free control diet, 2) control diet + 10% guar gum and 3) control diet + 10% pectin. In germ-free rats, guar gum and pectin had no effect on cecal polyamine concentrations. Putrescine was confirmed to be the major endogenous polyamine within the gut lumen. In cecal contents of conventional rats, both guar gum and pectin led to the appearance of cadaverine and to elevated putrescine concentrations in comparison with the fiber-free control diet (1.35 +/- 0.15 and 2.27 +/- 0.32, respectively, vs. 0.20 +/- 0.03 micromol/g dry weight, P < 0.05). The cecal cadaverine concentration was higher in pectin- than in guar-fed rats (8.20 +/- 0.89 vs. 1.92 +/- 0.27 micromol/g dry weight, P < 0.05). Counts of total bacteria, bacteroides, fusobacteria and enterobacteria were higher (P < 0.05) in rats fed guar gum and pectin. Bifidobacteria were found exclusively in guar-fed rats. In vitro studies on selected species representing the numerically dominant population groups of the human gut flora (bacteroides, fusobacteria, anaerobic cocci and bifidobacteria) were examined for their ability to synthesize intracellular polyamines. These experiments demonstrated the ability of bacteroides, fusobacteria and anaerobic cocci to synthesize high amounts of putrescine and spermidine. Calculations based on these results suggest that the intestinal microflora are a major source of polyamines in the contents of the large intestine.
Assuntos
Bactérias/metabolismo , Fibras na Dieta/farmacologia , Intestinos/microbiologia , Poliaminas/metabolismo , Animais , Bactérias/crescimento & desenvolvimento , Bacteroides/crescimento & desenvolvimento , Bacteroides/metabolismo , Cadaverina/biossíntese , Ceco/metabolismo , Ceco/microbiologia , Contagem de Colônia Microbiana , Enterobacteriaceae/crescimento & desenvolvimento , Enterobacteriaceae/metabolismo , Fusobacterium/crescimento & desenvolvimento , Fusobacterium/metabolismo , Galactanos/farmacologia , Vida Livre de Germes , Masculino , Mananas/farmacologia , Pectinas/farmacologia , Gomas Vegetais , Putrescina/biossíntese , Distribuição Aleatória , Ratos , Ratos WistarRESUMO
Polyamine synthesis is required in normal or neoplastic tissues if they are to continue to grow or divide. The highly inducible enzyme ornithine decarboxylase (ODC) catalyzes the conversion of ornithine to putrescine as the initial step in polyamine biosynthesis. The level of substrate pools of ornithine in cultured cells has been reported to markedly alter mitogen-induced ODC activity, putrescine accumulation, and DNA synthesis (V. Wu and C. V. Byus, Biochim. Biophys. Acta, 804: 89-99, 1984; V. Wu et al., Cancer Res., 41: 3384-3391, 1981). We attempted to limit the amount of ornithine available for polyamine biosynthesis in an animal by using a dietary approach. Since arginine serves as one of the intermediate biosynthetic precursors of ornithine, female CD-1 mice were placed on a special synthetic amino acid diet deficient in arginine. The ability of this arginine-free diet to alter epidermal ornithine and polyamine metabolism and tumorigenesis was assessed in the mouse two-stage model of skin carcinogenesis. The basal level of ornithine in the epidermis in control animals receiving the amino acid complete diet was very high compared to other tissues (155 nmol/mg protein). However, when the mice were fed the isocaloric arginine-free diet for a 2-week period, the levels of epidermal ornithine and arginine decreased by 40% (P less than 0.01). This reduction was blocked by the addition of 2% ornithine to the drinking water of the arginine-restricted animals. Acute administration of 12-O-tetradecanoyl-phorbol-13-acetate (TPA) to the epidermis caused a transient (4 and 8 h) reduction in ornithine and arginine but not lysine in the animals receiving the control, and ornithine-supplemented diets. The animals fed the special arginine-free diet exhibited a 40-50% reduction in tumor multiplicity or papillomas/mouse (P less than 0.05) and had a significantly lower tumor incidence or percentage of animals with tumour throughout a 19-week promotion period (P less than 0.02). However, the major effect of arginine restriction was consistent with an increase in tumor latency. The addition of ornithine completely reversed the reduction in the rate and extent of tumorigenesis in the arginine-free animals. The accumulation of putrescine (but not spermidine or spermine) in the epidermis following a single administration of TPA was significantly reduced in the animals receiving the arginine-free diet. The papillomas or tumors from the animals deprived of arginine had markedly reduced (less than 35%) levels of putrescine compared to the tumors from control animals, and appeared to be more sensitive to dietary arginine restriction than was the chronically promoted but untransformed epidermis.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Arginina/administração & dosagem , Ornitina Descarboxilase/biossíntese , Ornitina/biossíntese , Putrescina/biossíntese , Neoplasias Cutâneas/induzido quimicamente , Pele/metabolismo , 9,10-Dimetil-1,2-benzantraceno , Animais , Arginina/metabolismo , DNA/biossíntese , Feminino , Camundongos , Papiloma/induzido quimicamente , Poliaminas/metabolismo , Neoplasias Cutâneas/metabolismo , Acetato de TetradecanoilforbolRESUMO
Effects were examined of inanition, dietary aflatoxin (2.5 mg/kg), and dietary supplements of threonine, lysine, and arginine on the activities of renal arginase and hepatic ornithine decarboxylase and on the accumulation of polyamines in liver and brain of 24 or 26-day-old broiler cockerels. Aflatoxicosis and inanition lowered the activity of renal arginase by 58 and 37%, respectively. Supplemental dietary threonine (.4%) did not suppress the activity of renal arginase, while fortification of diets of controls with lysine (.53%), but not diets containing aflatoxin, elevated the activity of renal arginase. Supplements of dietary lysine and/or arginine did not influence the hepatic content of putrescine but lowered the concentrations of spermidine and spermine. Aflatoxicosis, but not inanition, increased the activity of hepatic ornithine decarboxylase (ODC; 22-fold), increased hepatic concentrations of putrescine and spermidine, but decreased spermine concentrations. The elevation of hepatic ODC, putrescine, and the ratio of spermidine to spermine parallels the enlargement of the liver caused by aflatoxicosis. Cadaverine and putrescine were not detected in avian brain, while cerebral concentrations of spermidine and spermine were not altered by aflatoxin, inanition, or by supplements of dietary lysine, arginine, or both lysine and arginine.
Assuntos
Aflatoxinas/intoxicação , Galinhas/metabolismo , Poliaminas/metabolismo , Doenças das Aves Domésticas/induzido quimicamente , Animais , Arginase/metabolismo , Arginina/farmacologia , Encéfalo/metabolismo , Rim/enzimologia , Fígado/enzimologia , Fígado/metabolismo , Lisina/farmacologia , Masculino , Ornitina Descarboxilase/metabolismo , Doenças das Aves Domésticas/enzimologia , Doenças das Aves Domésticas/metabolismo , Putrescina/biossíntese , Espermidina/biossíntese , Espermina/biossíntese , Treonina/farmacologiaRESUMO
The putrescine biosynthetic enzyme agmatine ureohydrolase (AUH) (agmatinase; EC 3.5.3.11) catalyzes the conversion of agmatine to putrescine in Escherichia coli. The specific activity of AUH was determined in crude extracts prepared from wild-type strains and from strains with mutations in the adenylate cyclase gene (cya) or the cAMP receptor protein gene (crp) or both. In glucose minimal medium, a delta cya strain exhibited 70 to 90% higher AUH activity than a cya+ strain. Addition of 1 to 10 mM cAMP to cya+ and delta cya strains cultured in glucose repressed AUH activity in a dose-dependent manner. Addition of 1 to 10 mM cAMP to a delta crp strain failed to repress AUH activity. Addition of agmatine resulted in a three- to fourfold induction of AUH in delta cya and delta crp strains. This induction could be blocked by the addition of chloramphenicol. Simultaneous additions of various proportions of cAMP and agmatine resulted in reduced levels of induction and repression of AUH activity. This antagonistic regulation was shown to be exerted by independent mechanisms since AUH activity could be induced by agmatine in a delta crp strain supplemented with cAMP. These results suggest that both agmatine and cAMP antagonistically regulate AUH activity at the level of transcription. In minimal medium supplemented with 1 mM putrescine, the strains did not exhibit repression of AUH activity. In contrast, in minimal medium supplemented with 1 mM ornithine or arginine, cya+ or delta cya strains exhibited induced AUH activity as a result of conversion of these substrates to agmatine. Further experiments in vitro demonstrated that the effects observed with cAMP, agmatine, and arginine were not post-translationally mediated.
Assuntos
Agmatina/farmacologia , AMP Cíclico/farmacologia , Escherichia coli/enzimologia , Guanidinas/farmacologia , Putrescina/biossíntese , Transcrição Gênica/efeitos dos fármacos , Ureo-Hidrolases/genética , Arginina/farmacologia , Cloranfenicol/farmacologia , Indução Enzimática , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Cinética , Ornitina/farmacologia , Ureo-Hidrolases/metabolismoRESUMO
Previous reports from this laboratory have shown that supplementation of diets containing the optimal level (0.02%) of dietary inorganic sulfate (SO4(2-] with cysteine instead of methionine can affect several metabolic pathways. It is possible that these results reflect alterations in the biosynthesis of potent physiological compounds, the polyamines. Adult male albino rats were fed diets containing 15% casein and a constant level of inorganic sulfate (0.02%) supplemented with cysteine (0.505%) or methionine (0.62%). The polyamines (putrescine, spermidine and spermine) and the controlling enzymes for their biosynthesis ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (SAMD) were evaluated in liver, kidney and brain tissue homogenates following a 17-day dietary period. Rats fed the diet supplemented with cysteine had increased ODC activity and decreased SAMD activity when compared to rats fed diets supplemented with methionine. Polyamine concentrations varied in tissues with a trend toward increasing amounts in animals fed the cysteine-supplemented diet. Based on these data, it appears that dietary cysteine stimulates the biosynthesis and increased tissue concentration of polyamines.
Assuntos
Cisteína/farmacologia , Metionina/farmacologia , Poliaminas/biossíntese , Adenosilmetionina Descarboxilase/metabolismo , Animais , Encéfalo/metabolismo , Rim/metabolismo , Fígado/metabolismo , Masculino , Ornitina Descarboxilase/metabolismo , Putrescina/biossíntese , Ratos , Ratos Endogâmicos , Espermidina/biossíntese , Espermina/biossínteseRESUMO
The catabolism of glucose by the oral mixed bacteria results in a lowering of the pH whereas arginine degradation favours a rise. In the mouth, low and high levels of glucose cause different plaque pH conditions which, in turn, might affect the rate and mode of degradation of arginine. This possibility was examined in the suspended salivary-sediment system where these pH conditions can be simulated. With the pH, the metabolic parameters examined were arginine utilization, ammonia, carbon dioxide and putrescine formation, utilization of glucose and changes in levels of L(+)- and D(-)-lactic acid. At the lower glucose concentration, the pH rapidly fell and then slowly rose whereas, with the higher glucose level, the pH showed a greater fall and no subsequent rise. The more acidic pH conditions favoured by the higher glucose level inhibited arginine degradation and the appearance of its various end-products and intermediates. Arginine degradation with arginine-[U-14C] and paper chromatography-autoradiography showed successive appearance of citrulline, ornithine and putrescine and, depending upon the pH, some succinate. When the pH was held constant at several different values, arginine degradation was optimal when the pH was near neutrality. In supplementary experiments, arginine had little effect on the ability of the oral mixed bacteria to utilize glucose and produce and utilize lactic acid, whereas the arginine peptide, arginylisoleucine and saliva supernatant stimulated these processes. Thus glycolysis enhancement and a more rapid clearance of fermentable carbohydrate by the oral bacteria would accompany pH-rise activity with arginine peptide and saliva but would not accompany pH-rise activity with arginine.