Alcohol oxidation by Candida guilliermondii yeasts grown on hexadecanol

A number of enzyme systems involved in the first steps of hexadecane oxidation can be induced by hexadecanol, an intermediate product of hexadecane degradation. It has also been found that, in Candida guilliermondii cells and in their mitochondrial fraction, an oxidase system is induced when the cells are grown on hexadecanol. This system is similar to that in cells grown on hexadecane; it oxidises higher alcohols at a high rate and is not inhibited by the inhibitors of the man phosphorylating respiration chain. The membrane-bound alcohol dehydrogenase and aldehyde dehydrogenase activities resistant to pyrazole, an inhibitor of cytosol ethanol dehydrogenase, are induced together with the oxidase activity when the cells are grown on hexadecanol as well as on hexadecane. The oxidation of higher alcohols by whole cells is entirely inhibited by azide although their oxidation by mitochondria is resistant to the action of azide; apparently, azide inhibits the transport of alcohols into the cell.     

The alkane-hydroxylating enzyme system of the yeast Candida guilliermondii.

The growth of the investigated Candida guilliermondii strain on n-alkanes induces an alkane-hydroxylating enzyme system, which consists of a cytochrome P-450 and a NADPH-dependent reductase. The cytochrome P-450 was purified to 4 nmoles per mg protein. Long-chain alkanes, preferably hexadecane to octadecane, are hydroxylated to the corresponding primary alcohol by this enzyme system. The substrate induces a type I spectrum, other compounds checked type II spectra.     

Effect of polyene antibiotics and their perhydrovderivatives on intact cells and protoplasts of yeast Candida guilliermondii]

Perhydroderivatives of polyene antibiotics have a much lower activity against eukaryotic cells than the polyene antibiotics itself. Bacterial cells are normally resistant against most polyene antibiotics and their perhydroderivatives. In earlier experiments with wall less L-form cells of Escherichia coli we have shown that the bacterial cell wall may be responsible for the resistance of the intact bacterial cells against polyene antibiotics and their perhydroderivatives by masking internal target sites. In the present paper we studied the effect of polyene antibiotics and their perhydroderivatives on intact cells and protoplasts of Candida guilliermondii. Our experiments have shown that most of the perhydroderivatives studied had a lower activity against intact cells as well as protoplasts than the corresponding polyene antibiotics. This means that in the case of eukaryotic cells the cell wall as a penetration barrier cannot mainly be responsible for the low activity of perhydroderivatives. The results are compared with those obtained previously with intact cells and protoplast type L-form cells of E. coli.     

Cytochrome P-450 and alkane hydroxylase activity in Candida guilliermondii.

In the investigated Candida guilliermondii strain after growth on n-alkanes as the only carbon and energy source 5--10 nMol cytochrome P-450 per g cells (wet weight) could be detected. Cytochrome P-450 and alkane hydroxylase activity was found in the 100 000 xg pellet. Cofactor studies and inhibition experiments revealed the existence of a NADPH-dependent cytochrome P-450 alkane hydroxylase system.     

The antilysozyme activity of yeasts in the genus Candida

The fungi of the genus Candida isolated from patients with oral mucosa candidiasis and from candidiasis carriers have been studied for their antilysozyme activity. These fungi (Candida albicans, C. tropicalis, C. krusei, C. quilliermondii) are antilysozyme-active. A high antilysozyme activity of the fungi isolated from patients with oral mucosa candidiasis permits supposing that the presence of this trait may be one of the factors of microorganism pathogenicity. The effective antimycotic therapy (clotrimasole, sanguirhitrin) decreases the antilysozyme activity of fungi of the genus Candida.     

Capsaicin and its analogs inhibit the activity of NADH-coenzyme Q oxidoreductase of the mitochondrial respiratory chain

Capsaicin and its analogs with different acyl moieties were found to inhibit the electron-transfer activity of NADH-coenzyme Q oxidoreductase isolated from beef heart mitochondria. The inhibitory potency of capsaicin was lower than those of dihydrocapsaicin and analogs with heptanoyl, capryl, undecanoyl, and lauroyl moieties, but was higher than those of analogs with palmitoyl and stearoyl moieties. The analog with the lauroyl moiety showed the strongest inhibition. These results suggest that hydrophobicity and the appropriate carbon chain length of the acyl moiety are important for the binding of compounds to the enzyme. On the other hand, capsaicin and its analogs did not interrupt rotenone-insensitive electron transfer from NADH to menadione. Furthermore, these compounds had almost no effect on the spectral properties and EPR signals arising from iron-sulfur clusters of the NADH-treated enzyme. Kinetic analyses with double-reciprocal plots showed that these compounds were competitive inhibitors with respect to coenzyme Q1, an electron acceptor. These results strongly suggest that capsaicin and its analogs bind to the coenzyme Q1 binding site of the enzyme.     

Effect of phosphate buffer concentration on the batch xylitol production by Candida guilliermondii

AIMS: To evaluate the effect of phosphate buffer concentration on growth and xylitol production by Candida guilliermondii FTI 20037. METHODS AND RESULTS: Fermentations runs were carried out in batch mode employing semisynthetic medium supplemented with phosphate buffer at different concentrations (from 200 to 600 mmol l(-1)). The xylitol yield (Y(P/S)) and volumetric productivity (Q(P)) were improved when the fermentation medium was supplemented with phosphate buffer at concentration of 600 mmol l(-1). Under this condition (Y(P/S)) and (Q(P)) values were 0.75 g g(-1) and 0.66 g l(-1) h(-1), respectively, whereas in the absence of the phosphate buffer these values decreased to 0.52 g g(-1) and 0.44 g l(-1)h(-1) respectively. CONCLUSIONS: The use of phosphate buffer at 600 mmol l(-1) promoted an easier pH control during shake flasks fermentation of C. guilliermondii. In addition the xylitol yield and productivity were significantly improved in response to the supplementation of potassium phosphate in the medium. The increase in these parameters could be related to both osmotic effect and pH control. SIGNIFICANCE AND IMPACT OF THE STUDY: This approach provided a method for improving the xylitol production from semisynthetic medium by C. guilliermondii, being possible their use as a simple strategy to achieve efficient fermentation processes employing complex medium such as lignocellulosic hydrolysates.     

Influence of the chain length of ubiquinones on their interaction with DPPC in mixed monolayers

The thermodynamic behavior of representative short (UQ2), middle (UQ4 and UQ6) and long-chain (UQ10) ubiquinones (UQ) mixed with dipalmitoyl-phosphatidylcholine (DPPC) was studied in monolayers at the air-water interface. The influence of isoprenoid chain-length of UQ on miscibility of both lipids was investigated by analysis of surface pressure-area isotherms and using fluorescence microscopy. Analysis of excess areas (A(ex)) and free energies of mixing (DeltaGm), calculated from compression isotherms in the full range of ubiquinones concentrations, has given evidences for UQ-rich constant-size (UQ6, UQ10) or less growth limited (UQ2, UQ4) microdomains formation within mixed films. Fluorescence microscopy observation revealed that ubiquinones are preferentially soluble in the expanded phase. When lateral pressure increased, concomitant evolutions of A(ex) and DeltaGm parameters, and composition dependence of collapse surface pressures, argue for an evolution towards a total segregation, never reached due to expulsion of ubiquinones from the film. The possible significance of these observations is discussed in relation to ubiquinones organization and similar chain length effects in membranes.     

Influence of the chain length of ubiquinones on their interaction with DPPC in mixed monolayers

The thermodynamic behavior of representative short (UQ2), middle (UQ4 and UQ6) and long-chain (UQ10) ubiquinones (UQ) mixed with dipalmitoyl-phosphatidylcholine (DPPC) was studied in monolayers at the air-water interface. The influence of isoprenoid chain-length of UQ on miscibility of both lipids was investigated by analysis of surface pressure-area isotherms and using fluorescence microscopy. Analysis of excess areas (A_ex) and free energies of mixing (ΔG_m), calculated from compression isotherms in the full range of ubiquinones concentrations, has given evidences for UQ-rich constant-size (UQ6, UQ10) or less growth limited (UQ2, UQ4) microdomains formation within mixed films. Fluorescence microscopy observation revealed that ubiquinones are preferentially soluble in the expanded phase. When lateral pressure increased, concomitant evolutions of A_ex and ΔG_m parameters, and composition dependence of collapse surface pressures, argue for an evolution towards a total segregation, never reached due to expulsion of ubiquinones from the film. The possible significance of these observations is discussed in relation to ubiquinones organization and similar chain length effects in membranes.     

Effect of beta-endorphin, enkephalins and their synthetic analogs on the neuronal electrical activity of the respiratory center in the medulla oblongata

It has been shown in experiments on conscious rabbits that beta-endorphine, enkephalins and their synthetic analogs as well as morphine suppress respiration depending on the dose. Naloxone completely reverses this effect of the drugs. While studying the mechanism of the suppressing action of morphine-like substances on respiration in experiments on anesthesized rabbits and cats, opioid peptides and morphine were applied microiontophoretically to the neurons of the bulbar respiratory center. These cells were found to be highly sensitive to the drugs (about 60% of both respiratory and reticular neurons were suppressed by microiontophoretic application of the drugs). Naloxone prevented the effects of opioids and morphine. It is assumed that the suppressing effect of endogenous opioid peptides and their synthetic analogs on respiration is determined to a considerable degree by direct influence of morphine-like substances on the neurons of the bulbar respiratory center.