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1.
Rev Argent Microbiol ; 31(2): 65-71, 1999.
Artigo em Espanhol | MEDLINE | ID: mdl-10425661

RESUMO

Data concerning the effect of very low concentrations of metals on citric acid production by microorganisms, as well as on the activity of enzymes presumptively involved in the process, are confuse. The bulk of information was obtained mainly studying selected strains of Aspergillus niger. Information concerning other citric acid producer filamentous fungi, such as A. wentii, is scanty. In the present article we report the effect of different cations on the growth pattern of A. wentii P1 as well as on the related citric acid production and the activity of several enzymes. It was found that without any addition to the culture medium the fungus developed a pelleted form of growth, pellets being about 1.5 mm in diameter. The citric acid yield was about 90%. The addition of Cu2+ impaired the sugar uptake, as well as the production of citric acid and biomass. The uptake of sugar increased in the presence of Zn2+, and there was a marked increase of the biomass production, which could account for the low citric acid production. The addition of Fe2+ impaired the citric acid production and, as sulfate, the sugar uptake. The presence of Fe3+ markedly impaired the citric acid production and increased the sugar uptake. There is no agreement about the enzymes involved in the accumulation of citric acid by microorganisms. In spite of this, aconitase (Ac), isocitrate lyase (IL), isocitrate dehydrogenase NAD(+)-dependent (ICDH- NAD+) and isocitrate dehydrogenase NADP(+)-dependent (ICDH-NADP+) are often postulated as key enzymes. In our case, these enzymes were active during the standard fermentation, although with variations, particularly concerning Ac and IL. The behavior of enzymes might be different when tested in vivo or in vitro, mainly from the quantitative point of view. Nevertheless, the activity determined in vitro might give some indication concerning the effect on fermentation of substances present in the medium. It was found that all the enzymes tested increased their activity in the presence of Fe2+ and Fe3+. The addition of Cu2+ improved the activity of Ac, IL and ICDH-NADP+, while that of ICDH-NAD+ was impaired. The presence of Zn2+ decreased the activity of the enzymes excepting that of ICDH-NADP+ which increased. It must be pointed out that according to these results there are differences concerning the behavior of ICDH-NAD+ and ICDH-NADP+ in the presence of Cu2+ or Zn2+.


Assuntos
Aspergillus/efeitos dos fármacos , Ácido Cítrico/metabolismo , Oligoelementos/farmacologia , Aconitato Hidratase/metabolismo , Aspergillus/enzimologia , Aspergillus/crescimento & desenvolvimento , Aspergillus/metabolismo , Proteínas de Bactérias/análise , Metabolismo dos Carboidratos , Cobre/farmacologia , Fermentação/efeitos dos fármacos , Ferro/farmacologia , Isocitrato Desidrogenase/metabolismo , Isocitrato Liase/metabolismo , Isoenzimas/metabolismo , Zinco/farmacologia
2.
Rev. argent. microbiol ; 31(2): 65-71, abr.-jun. 1999.
Artigo em Espanhol | LILACS | ID: lil-333171

RESUMO

Data concerning the effect of very low concentrations of metals on citric acid production by microorganisms, as well as on the activity of enzymes presumptively involved in the process, are confuse. The bulk of information was obtained mainly studying selected strains of Aspergillus niger. Information concerning other citric acid producer filamentous fungi, such as A. wentii, is scanty. In the present article we report the effect of different cations on the growth pattern of A. wentii P1 as well as on the related citric acid production and the activity of several enzymes. It was found that without any addition to the culture medium the fungus developed a pelleted form of growth, pellets being about 1.5 mm in diameter. The citric acid yield was about 90. The addition of Cu2+ impaired the sugar uptake, as well as the production of citric acid and biomass. The uptake of sugar increased in the presence of Zn2+, and there was a marked increase of the biomass production, which could account for the low citric acid production. The addition of Fe2+ impaired the citric acid production and, as sulfate, the sugar uptake. The presence of Fe3+ markedly impaired the citric acid production and increased the sugar uptake. There is no agreement about the enzymes involved in the accumulation of citric acid by microorganisms. In spite of this, aconitase (Ac), isocitrate lyase (IL), isocitrate dehydrogenase NAD(+)-dependent (ICDH- NAD+) and isocitrate dehydrogenase NADP(+)-dependent (ICDH-NADP+) are often postulated as key enzymes. In our case, these enzymes were active during the standard fermentation, although with variations, particularly concerning Ac and IL. The behavior of enzymes might be different when tested in vivo or in vitro, mainly from the quantitative point of view. Nevertheless, the activity determined in vitro might give some indication concerning the effect on fermentation of substances present in the medium. It was found that all the enzymes tested increased their activity in the presence of Fe2+ and Fe3+. The addition of Cu2+ improved the activity of Ac, IL and ICDH-NADP+, while that of ICDH-NAD+ was impaired. The presence of Zn2+ decreased the activity of the enzymes excepting that of ICDH-NADP+ which increased. It must be pointed out that according to these results there are differences concerning the behavior of ICDH-NAD+ and ICDH-NADP+ in the presence of Cu2+ or Zn2+.


Assuntos
Ácido Cítrico/metabolismo , Aspergillus , Oligoelementos , Aconitato Hidratase , Aspergillus , Carboidratos , Cobre , Fermentação/efeitos dos fármacos , Ferro , Isocitrato Desidrogenase , Isocitrato Liase , Isoenzimas , Proteínas de Bactérias/análise , Zinco
3.
Rev. argent. microbiol ; 31(2): 65-71, abr.-jun. 1999.
Artigo em Espanhol | BINACIS | ID: bin-6662

RESUMO

Data concerning the effect of very low concentrations of metals on citric acid production by microorganisms, as well as on the activity of enzymes presumptively involved in the process, are confuse. The bulk of information was obtained mainly studying selected strains of Aspergillus niger. Information concerning other citric acid producer filamentous fungi, such as A. wentii, is scanty. In the present article we report the effect of different cations on the growth pattern of A. wentii P1 as well as on the related citric acid production and the activity of several enzymes. It was found that without any addition to the culture medium the fungus developed a pelleted form of growth, pellets being about 1.5 mm in diameter. The citric acid yield was about 90. The addition of Cu2+ impaired the sugar uptake, as well as the production of citric acid and biomass. The uptake of sugar increased in the presence of Zn2+, and there was a marked increase of the biomass production, which could account for the low citric acid production. The addition of Fe2+ impaired the citric acid production and, as sulfate, the sugar uptake. The presence of Fe3+ markedly impaired the citric acid production and increased the sugar uptake. There is no agreement about the enzymes involved in the accumulation of citric acid by microorganisms. In spite of this, aconitase (Ac), isocitrate lyase (IL), isocitrate dehydrogenase NAD(+)-dependent (ICDH- NAD+) and isocitrate dehydrogenase NADP(+)-dependent (ICDH-NADP+) are often postulated as key enzymes. In our case, these enzymes were active during the standard fermentation, although with variations, particularly concerning Ac and IL. The behavior of enzymes might be different when tested in vivo or in vitro, mainly from the quantitative point of view. Nevertheless, the activity determined in vitro might give some indication concerning the effect on fermentation of substances present in the medium. It was found that all the enzymes tested increased their activity in the presence of Fe2+ and Fe3+. The addition of Cu2+ improved the activity of Ac, IL and ICDH-NADP+, while that of ICDH-NAD+ was impaired. The presence of Zn2+ decreased the activity of the enzymes excepting that of ICDH-NADP+ which increased. It must be pointed out that according to these results there are differences concerning the behavior of ICDH-NAD+ and ICDH-NADP+ in the presence of Cu2+ or Zn2+.(AU)


Assuntos
Estudo Comparativo , RESEARCH SUPPORT, NON-U.S. GOVT , Aspergillus/efeitos dos fármacos , Ácido Cítrico/metabolismo , Oligoelementos/farmacologia , Aconitato Hidratase/metabolismo , Aspergillus/enzimologia , Aspergillus/crescimento & desenvolvimento , Aspergillus/metabolismo , Proteínas de Bactérias/análise , Carboidratos/metabolismo , Cobre/farmacologia , Fermentação/efeitos dos fármacos , Ferro/farmacologia , Isocitrato Desidrogenase/metabolismo , Isocitrato Liase/metabolismo , Isoenzimas/metabolismo , Zinco/farmacologia
4.
Rev. argent. microbiol ; 31(2): 65-71, 1999 Apr-Jun.
Artigo em Espanhol | BINACIS | ID: bin-39994

RESUMO

Data concerning the effect of very low concentrations of metals on citric acid production by microorganisms, as well as on the activity of enzymes presumptively involved in the process, are confuse. The bulk of information was obtained mainly studying selected strains of Aspergillus niger. Information concerning other citric acid producer filamentous fungi, such as A. wentii, is scanty. In the present article we report the effect of different cations on the growth pattern of A. wentii P1 as well as on the related citric acid production and the activity of several enzymes. It was found that without any addition to the culture medium the fungus developed a pelleted form of growth, pellets being about 1.5 mm in diameter. The citric acid yield was about 90


. The addition of Cu2+ impaired the sugar uptake, as well as the production of citric acid and biomass. The uptake of sugar increased in the presence of Zn2+, and there was a marked increase of the biomass production, which could account for the low citric acid production. The addition of Fe2+ impaired the citric acid production and, as sulfate, the sugar uptake. The presence of Fe3+ markedly impaired the citric acid production and increased the sugar uptake. There is no agreement about the enzymes involved in the accumulation of citric acid by microorganisms. In spite of this, aconitase (Ac), isocitrate lyase (IL), isocitrate dehydrogenase NAD(+)-dependent (ICDH- NAD+) and isocitrate dehydrogenase NADP(+)-dependent (ICDH-NADP+) are often postulated as key enzymes. In our case, these enzymes were active during the standard fermentation, although with variations, particularly concerning Ac and IL. The behavior of enzymes might be different when tested in vivo or in vitro, mainly from the quantitative point of view. Nevertheless, the activity determined in vitro might give some indication concerning the effect on fermentation of substances present in the medium. It was found that all the enzymes tested increased their activity in the presence of Fe2+ and Fe3+. The addition of Cu2+ improved the activity of Ac, IL and ICDH-NADP+, while that of ICDH-NAD+ was impaired. The presence of Zn2+ decreased the activity of the enzymes excepting that of ICDH-NADP+ which increased. It must be pointed out that according to these results there are differences concerning the behavior of ICDH-NAD+ and ICDH-NADP+ in the presence of Cu2+ or Zn2+.

5.
Biotechnol Appl Biochem ; 26(3): 183-7, 1997 12.
Artigo em Inglês | MEDLINE | ID: mdl-9428156

RESUMO

After a 10.5-fold purification, beta-N-acetyl-D-glucosaminidase (EC 3.2.1.52) produced by Aspergillus niger 419, showed the following main characteristics: maximum activity at 65 degrees C, pH 4.5; K(m) and kcat using p-nitrophenyl-N-acetyl-beta-D-glucosaminide as substrate, 0.2 mM and 0.93 x 10(4) min-1, respectively; Ea, 30.5 kJ/mol; molecular mass, 131,000 Da; pI 4.4. The activity after heating for 15 min at 70, 75 and 80 degrees C was 70, 28 and 13% of that found at 65 degrees C, respectively. The enzyme was active in reaction mixtures containing glycerol, ethanol, methanol, propan-2-ol, acetone or dioxan. The presence of Sr2+ or Ca2+ enhanced the activity, while it was inhibited by Cu2+ and Fe3+. The enzyme was highly specific for p-nitrophenyl N-acetyl-beta-D-glucosaminide and no activity was found when p-nitrophenyl derivatives of N-acetyl-beta-D-galactosaminide, beta-D-galactopyranoside and beta-D-N,N'-diacetylchitobiose were tested as substrates. Due to its thermostability, specificity and resistance to different organic solvents, the enzyme might be a potentially useful tool for the analysis and production of oligosaccharides.


Assuntos
Acetilglucosaminidase/isolamento & purificação , Acetilglucosaminidase/metabolismo , Aspergillus niger/enzimologia , Acetilglucosaminidase/química , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Cinética , Peso Molecular , Especificidade por Substrato , Temperatura
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