Utilization of n-Alkanes by Pullularia pullulans

Pullularia pullulans was tested for its ability to utilize a series of n-alkanes for growth. It utilized hydrocarbons containing higher C-numbers (13, 14, 16, and 18) to a greater degree than those containing lower numbers; in addition, an induction phenomenon was noted. Gas-liquid and thin-layer chromatography of ether extracts of the growth media revealed that oleic and palmitic acids were formed from tridecane, tetradecane, hexadecane, and octadecane.     

The Degradation of Gallic Acid and its Alkyl Esters by Pullularia pullulans

By elective techniques an isolate of Pullularia pullulans was obtained which possessed the ability to utilize gallic acid (3,4,5-trihydroxybenzoic acid) and its alkyl esters together with other aromatic substrates as the sole carbon source for growth in a mineral salts medium. Quantitative measurements were made of the disappearance of the gallates from culture media and of their metabolism by washed suspensions by the Warburg technique. The pattern of oxygen uptake with various substrates by suspensions adapted to gallic acid was examined in the light of Stanier's simultaneous adaptation postulates in an attempt to identify intermediates of gallic acid metabolism. As a result, β-ketoadipic acid was suspected of being an intermediate. A keto compound was isolated as its 2,4-dinitrophenylhydrazone during gallic acid metabolism but it was not β-ketoadipic acid or levulinic acid.     

Glycogen Synthetase from Pullularia pullulans

3,4-Di-O-acetyl-2,6-dideoxy-2-(2′,4′-dinitroanilino)-6-phthalimido-α-d-glucopyranosyl bromide (I) was prepared in a good yield from glucosamine hydrochloride. A modified Königs-Knorr condensation of this bromide with a 2-deoxysteptamine derivative afforded a neamine derivative (IX) and its diastereomer (X). These compounds, (IX) and (X), were identified by PMR spectroscopy after conversion into the corresponding N-acetyl derivatives, (XI) and (XII).     

Plasmalogens in the yeast Pullularia pullulans

Choline and ethanolamine phosphoglycerides have been found in plasmalogen form in P. pullilans. Plasmalogens had not been described in yeasts up to now.     

Lipids of Aureobasidium (Pullularia) pullulans]

Fractional composition of free and bound lipids was studied in Aureobasidium (Pullularia) pullulans 8 by preparative TLC on Silufol. Bound lipids contained a fraction (27.76 +/- 0.5%) of dark brown colour, similar to melanin. The composition of fatty acids was studied by GLC. The following fatty acids were identified and determined quantitatively: C12:0, C14:0, C15:0, C16:0, C18:0, C18:1+C15:2. The following fatty acids predominated in free and bound lipids: C16:0, C18:1+C18:2. The ratio between unsaturated and saturated fatty acids in all fractions of free and bound lipids was more than unity. The following parameters were determined for lipids; ester number (173.89 and 178.53); iodine number (44.1 and 33.10), and saponification number (181.17 and 206.03) (the values are given for free and bound lipids, respectively).     

Uptake of Glucose-1-14C by Pullularia pullulans

Pullularia pullulans cells were grown in a medium containing yeast extract, malt extract, glucose, and nutrient salts, in addition to glucose-1-(14)C. The lipids extracted from the cells were fractionated by use of a single column packed with silicic acid. Of the total radioactive carbon added to the culture medium, the neutral lipid fractions contained 24.8%, whereas the phospholipid portions contained only 2.1%. The largest amount (16.5%) of (14)C among the neutral lipids was found in the fraction containing the free sterols. Among the phospholipids, the largest amount (1.1%) was found in phosphatidylserine and phosphatidylethanolamine. The second largest amount (9.6%) of the total (14)C used was found in trehalose, followed by carbon dioxide (7.3%).     

Isolation and identification of trehalase from Pullularia pullulans

Trehalase has been isolated from Pullularia pullulans. The enzyme, which is specific for trehalose, was purified approximately 800-fold. The optimal pH was found to be 4.0 and the Michaelis dissociation constant, K(m), was determined to be 3.2 x 10(-3)m.     

Structural Uniformity of Pullulan Produced by Several Strains of Pullularia pullulans

Extracellular polysaccharides produced by 3 strains of Pullularia pullulans were fractionated by treating with cetyl trimethyl ammonium hydroxide into soluble and insoluble fractions, and the structure of the former fraction, i.e., pullulan, was studied. The yield and the ratio of 2 fractions varied widely according to the strains. But the structure of pullulan was found to be uniform irrespective of the strains used. All 3 samples of pullulan gave only glucose on complete acid hydrolysis, and 93~95% maltotriose and 5~7% maltotetraose after isoamylase (pullulanase) action. The ratio of α-1,4- to α-1,6-glucosidic linkages calculated from periodate oxidation data coincided very well with the value expected from the ratio of maltotriose to maltotetraose units. An evidence for the complete absence of branch structure in pullulan was presented from the results of hydrolysis by pullulan 4-glucanohydrolase.     

出芽短梗霉发酵产生普罗蓝糖的研究

对一株野生型的出芽短梗霉(Aureobasidium pullulans)Ft1和从Ft1出发经原生质体再生筛选出的菌株R45进行了摇瓶发酵产普罗蓝糖的比较研究,结果表明R45无论从形态,菌体生长情况,还是从普罗蓝糖的产量,黑色素的产生等方面都与亲株Ft1有明显的区别,说明R45是一株具有一定生产价值的变异菌株.     

Utilization of Carbon Sources by Pullularia pullulans for the Elaboration of Extracellular Polysaccharides

The yeastlike fungus Pullularia pullulans utilizes simple mono- and disaccharides both in the production of cell mass and the elaboration of extracellular polysaccharide. The utilization pattern of these sugars and the effect obtained by varying the pH of the medium are studied, and the ability of the organism to utilize and elaborate extracellular polysaccharides from noncarbohydrate sources is explored.     

Flocculant and Chemical Properties of a Polysaccharide from Pullularia pullulans

An extracellular polysaccharide, PP-floc, was synthesized from glucose by Pullularia pullulans (or Aureobasidium pullulans) in a pilot plant batch fermentor containing 175 liters of culture medium. At 58 h of fermentation, the concentration of PP-floc was 1.03 g/100 ml, giving a 25.8% conversion of initial glucose to polysaccharide. The flocculant activity of the culture medium increased during the fermentation process and reached its maximum at 50 h of culture age. Less PP-floc (0.33 lb/ton of slimes [approximately 149.7 g/0.907 t]) was required to give the same flocculant activity as a synthetic polymer of acrylamide, Separan NP-10 (0.5 lb/ton of slimes [approximately 226.8 g/0.907 t]), at all temperatures from 25 to 100 C. The degree of inactivation of PP-floc and Separan NP-10 at elevated temperatures was almost identical, and they were completely inactivated at about the same temperature (80 C). PP-floc also gave better compaction of slimes than Separan NP-10 at all temperatures tested. PP-floc was soluble in water and its specific optical rotation was [alpha](D) (25) + 194 degrees in water (c, 0.4). PP-floc contained 83.3% carbohydrate, 3.2% protein, and 8.1% water. Glucose was found to be the principal sugar monomer with traces (>5%) of galactose and mannose present. Structural studies on the fractions of purified polysaccharide by methylation and by periodate oxidation techniques prove that PP-floc is linear and composed of alpha-(1 --> 4) and alpha-(1 --> 6) glucopyranosyl units in the approximate ratio of 2:1. The action of pullulanase on crude PP-floc suggested the ordered arrangement of two consecutive alpha-(1 --> 4)-linked glucopyranosyl units flanked by alpha-(1 --> 6)-linked glucopyranose residues.     

Variability of cultures of Pullularia pullulans with different ploidy levels

As the level of ploidy rises in Pullularia pullulans, this causes an increase in the frequency of spontaneous and UV-induced auxotrophic mutants as well as mutants with a modified respiration activity while the frequency of morphological mutants decreases. The latter can arise as a result of recessive and dominant mutations. A higher frequency of morphological mutants in the haploid may be result of recessive mutations. It is likely that the frequency of dominant mutations increases in cultures with a higher level of ploidy since, as the difference between the frequency of UV-induced mutants and the frequency of spontaneous morphological mutants increases.     

Effects of Topical Anesthetics on Pullularia pullulans and Debaryomyces hansenii

The inhibitory effects of three topical anesthetics of various concentrations on the growth of Pullularia pullulans, Debaryomyces hansenii, and on pigment production by P. pullulans were investigated. The topical anesthetics were benoxinate hydrochloride, proparacaine hydrochloride, and tetracaine hydrochloride. In decreasing order, the inhibiting effects of the drugs on growth were benoxinate, tetracaine, and proparacaine for P. pullulans, and tetracaine, benoxinate, and proparacaine for D. hansenii. The pigment formation in P. pullulans was inhibited by the three drugs.     

Induction and catabolite repression of L-rhamnose dehydrogenase in Pullularia pullulans.

The growth of Pullularia pullulans on L-rhamnose (6-deoxy-L-mannose) as the sole carbon source induces the synthesis of L-rhamnose dehydrogenase, a nicotinamide adenine dinucleotide-dependent enzyme that catalyzes the oxidation of the deoxy sugar to L-rhamnonolactone. The enzyme induction is inhibited by cycloheximide, suggesting de novo synthesis. The presence of d-glucose (0.2%) or D-galactose (0.2%) simultaneously with the inducer in the induction medium produced 50% repression of dehydrogenase synthesis, but no effect was detected with D-fructose and D-mannose at the same concentration. High levels of D-glucose (2%), under maximal catabolite repression conditions, produced a complete inhibition of enzyme synthesis.