Efecto de metales traza sobre la morfología celular, actividad enzimßtica y producción de ácido cítrico en una cepa de Aspergillus wentii / Effect of trace metals on cell morphology, enzyme activation, and production of citric acid in a strain of Aspergillus wentii

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+.

Atividade da isocitrato liase em Mucor rouxii / Activity of isocitrate liase in mucor rouxii

O ficomiceto dimórfico Mucor rouxii foi cultivado em meio que continha glucose a 2% (YPG), tendo-se verificado um crescimento bifásico característico. A isocitrato liase, enzima do ciclo do glioxilato, apareceu sob estas condiçöes somente após a glucose do meio ter sido totalmente esgotada. Altos níveis de atividade da isocitrato liase foram encontrados durante o metabolismo gluconeogenético. Isso mostra a participaçäo do ciclo do glioxilato como fonte de succinato e de oxalacetato para as vias biossintéticas do fungo. Em meio complexo sem glucose (YP), estritamente gluconeogenético, foram encontradas altas atividades de isocitrato liase