Regulation of the central metabolism in relation to citric acid production in Saccharomycopsis lipolytica

The mechanism of the massive extracellular production of citric and isocitric acids by Saccharomycopsis lipolytica grown on n-paraffins has been studied. When growth stops, because of nitrogen limitation, the intracellular concentration of ATP sharply rises whereas that of AMP and ADP decreases to a low level. At the same time production of acids begins. The activity of the NAD-dependent isocitrate dehydrogenase which requires AMP for activity becomes very low and prevents the oxidative function of the citric acid cycle whereas isocitrate lyase is not inhibited. As citrate synthase inhibition by ATP appears to be insufficient to stop n-paraffin degradation, citric and isocitric acids accumulation can take place. Massive excretion of these acids, however, probably still involves other physiological changes brought about by nitrogen limitation, possibly some permeabilization of the cell to these acids.This work is a part of a Doctorat de Spécialité Thesis submitted by R. Marchal to the University of Nancy (1975)     

Spermatozoal morphologies and fructose and citric acid concentrations in agouti (Dasyprocta leporina) semen

This study was conducted to identify the levels of fructose and citric acid, and sperm morphologies in agouti (Dasyprocta leporina) semen. These parameters may be important in identifying highly fertile semen from the agouti. The objectives were: (1) to investigate spermatozoal abnormalities in agouti semen and (2) to determine the concentrations of seminal fructose and citric acid in agouti semen samples. Semen samples were collected from 16 anaesthetised male agouti by electro-ejaculation. Fructose and citric acid concentrations were 256.86+/-63.54 mg/dl and 1877+/-147 mg/dl, respectively, measured with ELISA kits. Sperm morphologies, examined using eosin-negrosin staining, showed 11 morphologies. The most abundant (68.5%) sperm morphology (M1) showed no known sperm defects. Means for head, mid piece, tail and total length of the agouti spermatozoa was 5.23+/-0.04 microm, 5.18+/-0.08 microm, 37.52+/-0.24 microm and 47.96+/-0.25 microm, respectively for M1 sperm. The means of spermatozoa head and mid piece width and semen volume were 3.26+/-0.04 microm, 0.70+/-0.02 microm and 0.47+/-0.16 ml, respectively. It was concluded that as the fructose concentration in agouti ejaculate increased the percentage of spermatozoa with known spermatozoa defects increased (r=0.506; P<0.037; n=32). It is suggested that the M1 sperm could be the most competitive spermatozoa in agouti ejaculate. In conclusion standards for identifying fertile agouti semen were established.     

Studies on some enzymes relevant to citric acid accumulation by Aspergillus niger

Enzymatic studies have been performed on a local strain of Aspergillus niger to find a correlation with citric acid accumulation. The activity of aconitase [aconitate hydratase, citrate(isocitrate) hydrolyase, EC 4.2.1.3] and isocitrate dehydrogenase (NADP⁺) [threo-d_s-isocitrate:NADP⁺ oxidoreductase (decarboxylating) EC 1.1.1.42] decreased after 4 days whereas that of citrate synthase [citrate oxaloacetate-lyase (pro-3S-CH₂COO^?→acetylCoA), EC 4.1.3.7] did so after 8 days, when citric acid accumulation in the medium reached a maximum (45.9 mg ml^?1). In vitro studies with mycelial cell-free extracts demonstrated inhibition of citrate synthase activity by sodium azide and potassium ferricyanide on both the 4th and 8th days. Aconitase was inhibited by sodium arsenate, sodium fluoride, iodoacetic acid and potassium ferricyanide only on the 4th day. Isocitrate dehydrogenase (NADP⁺) activity on the 4th and 8th days was inhibited by iodoacetic acid but was stimulated by potassium ferricyanide. The possible existence of isozyme species of these enzymes is discussed.