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C Ratledge

Publications and source records attributed to C Ratledge.

At least 91 records · Page 5Linked to original sources
PubMed1977

Pathways of glucose metabolism in Candida 107, a lipid-accumulating yeast.

Phosphofructokinase was not detected in extracts of Candida 107 prepared in a variety of ways but was highly active in cells treated with toluene. Disruption of these cells destroyed activity of phosphofructokinase indicating that the enzyme is extremely labile. As patterns of labelling from [I-14C]glucose and [6-14C]glucose showed that 60% of glucose was metabolized via the pentose cycle, augmentation of this cycle is necessary to account for the high molar growth yields of this yeast. Phosphoketolases, reacting with xylulose 5-phosphate and fructose 6-phosphate, were found but the extent to which they contribute to glucose metabolism was not assessed.

Aldehyde-Lyases↗
PubMed1977

Lipid accumulation in an oleaginous yeast (Candida 107) growing on glucose in single-stage continuous culture.

Lipid accumulation of Candida 107, grown at dilution rates from 0.03 to the maximum of 0.21/h, with carbon, nitrogen, phosphate, and magnesium limitations in a chemostat, was maximal at about 40% (wt/wt) with nitrogen-limited medium at a dilution rate of 0.06/h, giving an efficiency of substrate conversion of 22 g of lipid per g of glucose consumed. At higher dilution rates the lipid content decreased. With carbon-limited growth, the highest lipid content (14%, wt/wt) was at the maximum dilution rate. High lipid contents also occurred with phosphate + nitrogen as double limitations of growth, with the lipid content of the yeast (about 35%, wt/wt) continuing to be near maximum at dilution rates also near maximum (0.17/h), thus giving the highest specific rate of lipid formation of any growth conditions (0.59 g of lipid/g of yeast per h). However, the efficiency of substrate utilization was only 5.2 g of lipid formed per 100 g of glucose consumed. The composition of the fatty acyl residues within the lipid remained constant over many weeks if the steady-state conditions remained unchanged. With carbon-limited growth, the degree of unsaturation of the fatty acids markedly decreased as the dilution rate was increased, but with nitrogen limitation the reverse trend was seen. In all cases, linoleic and oleic acids were the principal fatty acyl residues affected, and their relative proportions always varied in opposite directions. When magnesium was a limiting nutrient, there was a considerable increase in the proportion of myristic acid produced within the lipid. Neutral lipids (predominantly triglycerides) varied from 66 to 92% of the total lipid from carbon- and nitrogen-limited growth; phospholipids (varying from 2 to 25%) were highest in nitrogen-limited growth. The fatty acyl residues within each lipid fraction showed the same variations with changing growth rates.

Candida↗
PubMed1977

Lipid accumulation in an oleaginous yeast (Candida 107) growing on glucose under various conditions in a one- and two-stage continuous culture.

Lipid accumulation and fatty acid composition in Candida 107 have been studied using a two-stage continuous culture system in which the first vessel was run under carbon-limited conditions and then the entire output was passed into a second vessel, where lipid accumulation was stimulated by adding only glucose. Maximum lipid accumulation (28% of yeast [dry weight]) occurred for a volume ratio of vessel 1 to vessel 2 of 3:5, with 30 g of glucose per liter being added to vessel 2 operated at 25 degrees C with an aeration rate of between 0.1 and 1.0 volume of air/volume of medium per min. Although the maximum specific rate of lipid formation (0.05 g of lipid/g of yeast per h) was higher than in a nitrogen-limited, single-stage system, the efficiency of lipid formation was much less and never exceeded 14 g of lipid produced per 100 g of glucose consumed. The fatty acid composition was not significantly altered in either the two-stage or single-stage culture (nitrogen-limited) systems by changes in growth temperature (from 19 to 33 degrees C) or aeration rates (0.05 to 1.0 volume of air/volume of medium per min); or, in the two-stage system, by changes in the residence time of the yeast in the second vessel (from 3.2 to 24.4 h), or, in the single-stage system, by changes in pH (from 3.5 to 7.5). Only when the concentration of glucose entering vessel 2 of the two-stage system was less than 30 g/liter did significant changes in the fatty acids occur. Thus, although a two-stage continuous culture system allows lipid accumulation to be separated from the growth phase, it offers no practical advantages over a single-stage system as a means of producing microbial oils and fats.

Journal Article↗
PubMed1977

Oxygen demand by lipid-accumulating yeasts in continuous culture.

Specific oxygen uptake rates [Q(02)] were highest when Candida 107 and Rhodotorula gracilis grew at their maximum dilution rates. In nitrogen-limited medium, which encourages lipid accumulation, Q(02) values of both yeasts decreased as the dilution rate was lowered.

Candida↗
PubMed1976

Isolation, properties and taxonomic relevance of lipid-soluble, iron-binding compounds (the nocobactins) from Nocardia.

Representative strains of the genus Nocardia, when grown on iron-deficient media, produce intracellular lipid-soluble, iron-binding compounds known as nocobactins. However, strains representing the 'rhodochrous' taxon fail to form such compounds. The formation of the nocobactins is completely repressed in bacteria grown on iron-sufficient media. Procedures for purifying the nocobactins are described. From their various properties (u.v. and visible spectra of the the ferri- and desferri-materials, mobility upon thin-layer chromatography, and dissociation patterns in HCl), they can be distinguished from the mycobactins (related compounds from mycobacteria) and divided into three main classes. The classes correspond to the three well-described species of the genus Norcardia: N. asteroides, N. brasiliensis and N. caviae. The correlation of the nocobactin data with previous results of conventional numerical taxonomy is high. Two strains of N. asteroides, however, did not produce nocobactins of the group type.

Azepines↗
PubMed1975

The effect of p-aminosalicyclic acid on iron transport and assimilation in mycobacteria.

p-Aminosalicylic acid inhibits growth of Mycobacterium bovis BCG and Mycobacterium smegmatis more effectively if cells are growing with a sufficiency of iron (more than 1 mu g Fe/ml) in the medium than if cells are deficient in iron (smaller than 0.1 mu g Fe/ml). In iron-deficient cultures formation of mycobactin, an ionophore for iron transport, is strongly inhibited by p-aminosalicylic acid. Uptake of iron into cell suspensions is also inhibited and the activity of several iron-containing enzymes declines in cells exposed to p-aminosalicylic acid during their growth. p-Aminosalicylic acid is about 50 times more effective towards a mutant of M. smegmatis which required mycobactin under iron-deficient growth conditions than towards the wild-type parent. p-Aminosalicylate is taken up into cells by an active process independent of the salicylate uptake system, possibly by the route used for assimilation of p-aminobenzoate. (This could account for why p-aminobenzoic acid, but not salicylic acid, antagonizes the action of p-aminosalicylic acid.) With iron-deficient cells, salicylate assimilation is about 50 times greater than either p-aminosalicylate or p-aminobenzoate but with iron-sufficient cells and with the mycobactin mutant salicylate uptake is negligible whereas p-aminobenzoate and p-aminosalicylate uptakes are unaffected. p-Aminosalicylic acid at 3.3 mM (500 mu g/ml) partially inhibits the uptake of both p-aminobenzoate and, if it is occurring, that of salicylate as well. As p-aminosalicylic acid is always more effective when the intracellular concentration of salicylic acid is low, it probably acts as an anti-metabolite of salicylic acid, not, however, by inhibiting the conversion of salicylic acid to mycobactin, but probably somewhere along the metabolic pathway of iron uptake.

Aminobenzoates↗
PubMed1975

An analysis of intermediary metabolism and its control in a fat-synthesizing yeast (Candida 107) growing on glucose or alkanes.

Enzymes of glycolysis, pentose phosphate pathway, gluconeogenesis, tricarboxylate acid cycle, glyoxylate by-pass and fatty-acid biosynthesis were assayed in extracts from Candida 107 grown continuously on glucose under carbon limitation, nitrogen limitation and on n-alkanes. The yeast was therefore either in a lipogenic or lipolytic state. Phosphofructokinase was absent under all conditions whereas enzymes of gluconeogenesis, including fructose 1,6-bisphosphatase and the pentose phosphate cycle, were all present. Glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were specific for NADP+ and were inhibited in a non-competitive manner by NADPH and NADH. Phosphoenolpyruvate, citrate, ATP and acetyl CoA had no inhibitory effects. Thus glucose metabolism appears to be by the pentose phosphate pathway which will rapidly produce NADPH. This can readily be consumed during fatty-acid biosynthesis and, as there appears to be no inhibition of the flow of carbon from glucose to acetyl CoA, fatty-acid synthesis can continue for as long as there is a supply of glucose. These results help to explain the probable causes of fat build-up to high concentrations (about 40% of the cell dry weight) in this and other organisms. In alkane-grown cells, lipogenesis is repressed and carbon is able to flow from the alkanes via acetyl CoA, oxaloacetate and pyruvate into pentoses and hexoses in a unidirectional manner, because of the strong repression of pyruvate kinase and the increased activities of phosphoenolpyruvate kinase and fructose 1,6-biosphosphatase under these conditions. Although there was little change in the total activity of the TCA cycle enzymes under the various growth conditions, isocitrate lyase was induced under lipolytic conditions.

Acetyl Coenzyme A↗
PubMed1975

Extracellular iron acquisition by mycobacteria: role of the exochelins and evidence against the participation of mycobactin.

Mycobacterium bovis var. BCG was grown under iron-deficient conditions in the presence and absence of 1% Tween 80. Mycobactin, the iron iron ionophore of mycobacteria, was found solely within the bacteria grown in the absence of Tween, but low concentrations (0.75 mug/ml) of it appeared in the medium in the presence of the surfactant. Both types of medium contain agents, named exochelins, which could solubilize iron. 55Fe added to spent culture media was recovered only chelated to these compounds. Two exochelins were detected, isolated, and purified. Neither were precursors or breakdown products of mycobactin. In the desferri-form, exochelin MB-2, the major component, reversed the inhibitory effect of serum on the growth of BCG, and in their ferri-forms exochelins MB-1, MB-2, and MS (from Mycobacterium smegmatis) stimulated the growth of their producing organism in the presence of serum. Exochelin MB-2 could physically remove iron from ferritin, and BCG used ferritin as a source of iron during growth even when ferritin was separated from the bacteria by a dialysis membrane. As solutions of the exochelins were freely dialyzable, whereas solutions of mycobactin, even in Tween, were not, only exochelin could have been active in this experiment. The exochelins are proposed as the functional extracellular iron-binding agents of BCG and other mycobacteria, the role of mycobactin being confined to that of a cell wall iron transporter.

BCG Vaccine↗
PubMed1974

Isolation and structure of nocobactin NA, a lipid-soluble iron-binding compound from Nocardia asteroides.

Nocobactin NA, a lipid-soluble iron-chelating product with an unusual and characteristic u.v.-absorption spectrum, was isolated from Nocardia asteroides grown under conditions of iron deficiency. Its structure was determined by physical methods and by synthesis of one of its degradation products. Nocobactin NA was obtained as a homologous mixture of compounds with side chains of differing length, and resembles mycobactin M in structure except that it has an oxazole ring in place of an oxazoline ring, and the side chains in the cobactin fragment are considerably shorter.

Chemical Phenomena↗