Growth of Aureobasidium pullulans on straw hydrolysate.

Growth characteristics and cell properties of Aureobasidium (Pullularia) pullulans were studied. The organism grew well on an acid hydrolysate of ryegrass straw over a wide range of pH and temperature. The optimum temperature and pH for the growth of the organism were 32 degrees C and 5.5, respectively. A cell yield of 1.5 g/liter of straw hydrolysate was obtained. The dried cell mass contained 42.6% crude protein, 0.4% crude fat, and 6.4% nucleic acids. The essential amino acid profile of the microbial protein was comparable to that of Candida utilis. A rat feeding study indicated that the A. pullulans cells were not toxic and that the feed intake, weight gain, and protein efficiency ratio values were superior to those obtained with C. utilis. Once the question of mathogenicity is resolved, A. pullulans could be useful for production of single-cell protein from cellulosic wastes.     

Growth of Aureobasidium pullulans on straw hydrolysate.

Growth characteristics and cell properties of Aureobasidium (Pullularia) pullulans were studied. The organism grew well on an acid hydrolysate of ryegrass straw over a wide range of pH and temperature. The optimum temperature and pH for the growth of the organism were 32 degrees C and 5.5, respectively. A cell yield of 1.5 g/liter of straw hydrolysate was obtained. The dried cell mass contained 42.6% crude protein, 0.4% crude fat, and 6.4% nucleic acids. The essential amino acid profile of the microbial protein was comparable to that of Candida utilis. A rat feeding study indicated that the A. pullulans cells were not toxic and that the feed intake, weight gain, and protein efficiency ratio values were superior to those obtained with C. utilis. Once the question of mathogenicity is resolved, A. pullulans could be useful for production of single-cell protein from cellulosic wastes.     

Pullulan Elaboration by Aureobasidium pullulans Protoplasts

Protoplasts of Aureobasidium pullulans are capable of producing pullulan. Biosynthesis of the polymer pullulan required induction with kinetics similar to those of whole cells. The protoplasts also produced a heteropolysaccharide component containing mannose, glucose, and galactose. The relative proportions of the pullulan and heteropolysaccharide fractions were a function of glucose concentration, with the pullulan content of the total polysaccharide rising from 20% at 2.5 mM glucose to 45% at 20 mM glucose. Elaboration of pullulan by both cells and protoplasts was sensitive to 0.6 M KCl, which was present as the osmotic stabilizer in protoplast experiments. The presence of KCl resulted in a shift in the pH optimum to a more acidic value. The molecular weight of the protoplast-derived pullulan was sharply reduced from the molecular weight of the whole-cell-derived product. Exposure of the protoplasts to proteolytic enzymes had no effect on polysaccharide elaboration.     

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.     

Inorganic Sulfur Oxidation by Aureobasidium pullulans

Aureobasidium pullulans (de Bary) Arnaud isolated from the phylloplane of sycamore exposed to heavy atmospheric pollution oxidized S⁰ to S₂O₃²⁻, S₄O₆²⁻, and SO₄²⁻ in vitro. The intermediates S₂O₃²⁻ and S₄O₆²⁻ were also oxidized to SO₄²⁻. Cell-free extracts of A. pullulans also oxidized reduced forms of S, the oxidation increasing linearly with increasing protein concentration, showing that the process is enzymatic. The possible role of fungi in S oxidation in soils is discussed.     

出芽短梗霉胞外酸性漆酶

通过愈创木酚法平板检测10株出芽短梗霉,发现5株菌能够分泌胞外多酚氧化酶,反应最适pH在2.0左右,均属于酸性多酚氧化酶。菌株NG的酶活最高,达110 U/mL。添加H2O2、EDTA以及过氧化氢酶不显著影响菌株NG胞外酶活,表明NG分泌的多酚氧化酶中不含有锰过氧化物酶（MnP）和不依赖Mn2＋的过氧化物酶（MiP）,属于漆酶（Lac）。     

Growth and amylolytic activity of Aureobasidium pullulans in starch-limited culture

One strain of the yeast-like fungus Aureobasidium pullulans has been found which converts starch into biomass with a high yield (Y_starch = 0.590) and releases glucoamylase (1,4-α-d-glucan glucohydrolase EC 3.2.1.3) to a certain extent (ca. 2.2–2.3 U ml^?1) into the culture medium. The rate of starch hydrolysis seems to be high enough so as not to limit the specific growth rate.     

Biosynthesis of novel exopolymers by Aureobasidium pullulans

Aureobasidium pullulans ATCC 42023 was cultured under aerobic conditions with glucose, mannose, and glucose analogs as energy sources. The exopolymer extracts produced under these conditions were composed of glucose and mannose. The molar ratio of glucose to mannose in the exopolymer extract and the molecular weight of the exopolymer varied depending on the energy source and culture time. The glucose content of exopolymer extracts formed with glucose and mannose as the carbon sources was between 91 and 87%. The molecular weight decreased from 3.5 x 10(6) to 2.12 x 10(6) to 0.85 x 10(6) to 0.77 x 10(6) with culture time. As the culture time increased, the glucose content of the exopolymer extract formed with glucosamine decreased from 55 +/- 3 to 29 +/- 2 mol%, and the molecular weight increased from 2.73 x 10(6) to 4.86 x 10(6). There was no evidence that glucosamine was directly incorporated into exopolymers. The molar ratios of glucose to mannose in exopolymer extracts ranged from 87 +/- 3:13 +/- 3 to 28 +/- 2:72 +/- 2 and were affected by the energy source added. On the basis of the results of an enzyme hydrolysis analysis of the exopolymer extracts and the compositional changes observed, mannose (a repeating unit) was substituted for glucose, which gave rise to a new family of exopolymer analogs.     

Biosynthesis of Novel Exopolymers by Aureobasidium pullulans

Aureobasidium pullulans ATCC 42023 was cultured under aerobic conditions with glucose, mannose, and glucose analogs as energy sources. The exopolymer extracts produced under these conditions were composed of glucose and mannose. The molar ratio of glucose to mannose in the exopolymer extract and the molecular weight of the exopolymer varied depending on the energy source and culture time. The glucose content of exopolymer extracts formed with glucose and mannose as the carbon sources was between 91 and 87%. The molecular weight decreased from 3.5 × 10⁶ to 2.12 × 10⁶ to 0.85 × 10⁶ to 0.77 × 10⁶ with culture time. As the culture time increased, the glucose content of the exopolymer extract formed with glucosamine decreased from 55 ± 3 to 29 ± 2 mol%, and the molecular weight increased from 2.73 × 10⁶ to 4.86 × 10⁶. There was no evidence that glucosamine was directly incorporated into exopolymers. The molar ratios of glucose to mannose in exopolymer extracts ranged from 87 ± 3:13 ± 3 to 28 ± 2:72 ± 2 and were affected by the energy source added. On the basis of the results of an enzyme hydrolysis analysis of the exopolymer extracts and the compositional changes observed, mannose (a repeating unit) was substituted for glucose, which gave rise to a new family of exopolymer analogs.     

Pullulan production from deproteinized whey by Aureobasidium pullulans

Aureobasidium pullulans P56 was investigated using an adaptation technique and a mixed culture system. The adaptation of A. pullulans and the mixed cultures of A. pullulans and/or Lactobacillus brevisX20, Debaryomyces hansenii 194 and Aspergillus niger did not increase the production of polysaccharide. Enzymic hydrolysis of lactose in deproteinized whey gave a higher polysaccharide concentration and polysaccharide yield than acidic hydrolysed lactose. Maximum polysaccharide concentration (11.0 ± 0.5 g L⁻¹), biomass dry weight (10.5 ± 0.4 g L⁻¹), polysaccharide yield (47.2 ± 1.8%) and sugar utilization (93.2 ± 2.8%) were achieved using enzyme-hydrolysed whey (pH 6.5) containing 25 g L⁻¹ lactose and supplemented with K₂HPO₄ 0.5%, L-glutamic acid 1%, olive oil 2.5%, and Tween 80 0.5%. In this case the pullulan content of the crude polysaccharide was 40%. Received 16 December 1997/ Accepted in revised form 12 March 1999     

Pullulan production from brewery wastes by Aureobasidium pullulans

The production of pullulan from brewery wastes by Aureobasidium pullulans in shake flask culture was investigated. The maximum pullulan concentration (6.0g/l) was obtained after 72h of fermentation. The external addition of nutrients into the spent grain liquor improved significantly the production of pullulan. In this case, the highest values of pullulan concentration (11.0±0.5g/l), pullulan yield (48.2±1.5%), and sugar utilization (99.0±0.5%) were obtained in the medium (pH 6.5–7.5) supplemented with K₂HPO₄ 0.5%, l-glutamic acid 1%, olive oil 2.5%, and Tween 800.5%.     

Color Variants of Aureobasidium pullulans Overproduce Xylanase with Extremely High Specific Activity

Xylanase activity from naturally occurring color variants of Aureobasidium pullulans was associated with extracellular monomeric proteins of 20 to 21 kilodaltons. Xylanase represented nearly half the total extracellular protein, with a yield of up to 0.3 g of xylanase per liter. The specific activity of partially purified xylanase exceeded 2,000 IU/mg. Xylanase from typically pigmented strains appeared similar to that from color variants with respect to molecular weight, pH and temperature optima, and specific activity of purified (but not crude) enzyme. However, xylanase from typical strains made up only about 1.0% of total extracellular protein. Xylanase from strains of Cryptococcus albidus was associated with abundant proteins of about 43 kilodaltons and showed much lower specific activity.     

Purification and biochemical characterization of polygalacturonases produced by Aureobasidium pullulans

The extracellular polygalacturonases produced by Aureobasidium pullulans isolated from waters of the Danube river were partially purified and characterized. The pH optima of polygalacturonases produced in the first phases of cultivation (48 h) and after 10 d as well as their optima of temperature, thermal stabilities, molecular masses, isoelectric points, action pattern and ability to cleave polymeric and oligomeric substrates were compared. Polygalacturonases with a random action pattern (random cleavage of pectate forming a mixture of galactosiduronides with a lower degree of polymerization) [EC 3.2.1.15] were produced only in the first phases of growth, while exopolygalacturonases [EC 3.2.1.67] with a terminal action pattern (cleavage of pectate from the nonreducing end forming D-galactopyranuronic acid as a product) were found during the whole growth. The main enzyme form with a random action pattern was glycosylated and its active site had the arrangement described previously for the active site of polygalacturonase of phytopathogenic fungi.     

Biocontrol and Plant Growth Promotion Characterization of Bacillus Species Isolated from Calendula officinalis Rhizosphere

The phenotypic and genotypic diversity of the plant growth promoting Bacillus genus have been widely investigated in the rhizosphere of various agricultural crops. However, to our knowledge this is the first report on the Bacillus species isolated from the rhizosphere of Calendula officinalis. 15 % of the isolated bacteria were screened for their important antifungal activity against Fusarium oxysporum, Botrytis cinerea, Aspergillus niger, Cladosporium cucumerinium and Alternaria alternata. The bacteria identification based on 16S r-RNA and gyrase-A genes analysis, revealed strains closely related to Bacillus amyloliquefaciens, B. velezensis, B. subtilis sub sp spizezenii and Paenibacillus polymyxa species. The electro-spray mass spectrometry coupled to liquid chromatography (ESI-LC MS) analysis showed that most of the Bacillus isolates produced the three lipopeptides families. However, the P. polymyxa (18SRTS) didn’t produce any type of lipopeptides. All the tested Bacillus isolates produced cellulase but the protease activity was observed only in the B. amyloliquefaciens species (9SRTS). The Salkowsky colorimetric test showed that the screened bacteria synthesized 6–52 μg/ml of indole 3 acetic acid. These bacteria produced siderophores with more than 10 mm wide orange zones on chromazurol S. The greenhouse experiment using a naturally infested soil with Sclerotonia sclerotiorum showed that the B. amyloliquefaciens (9SRTS) had no significant (P > 0.05) effect on the pre-germination of the chickpea seeds. However, it increased the size of the chickpea plants and reduced the stem rot disease (P < 0.05).These results suggested that the Bacillus strains isolated in this work may be further used as bioinoculants to improve the production of C. officinalis and other crop systems.     

Exopolysaccharide biosynthesis by a fast-producing strain of Aureobasidium pullulans

Summary The mutant strain Aureobasidium pullulans ICCF-68 was able to produce in batch fermentation on a glucose medium of 80 g/l, exopolysaccharide at high volumetric productivity and final concentration (1.05 g/l.h and 50.2 g/l, respectively). A specific pH pattern and very high oxygen requirement were shown.     

Production of fructo-oligosaccharides from molasses by Aureobasidium pullulans cells

Three different strains of Aureobasidium pullulans were grown in batch cultures to compare their abilities for the production of fructo-oligosaccharides. Specific intracellular enzyme activity was the highest with strain KCCM 12017 and enzyme production was closely coupled to growth. Using A. pullulans cells, 166 g/l fructo-oligosaccharides was produced from 360 g/l molasses sugar as sucrose equivalent at 55 degrees C and pH 5.5 after 24 h incubation.     

Oxidation and reduction of lignin-related aromatic compounds by Aureobasidium pullulans

Summary A yeast-like fungus, identified as Aureobasidium pullulans, was isolated from a kraft mill settling pond by enrichment culture on 1-(4-hydroxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-propane-1,3-diol (I). The fungus was also able to use the following aromatic acids as sole carbon source: Benzoic, p-hydroxybenzoic, vanillic, syringic, ferulic and protocatechuic acids. Various aromatic alcohols were oxidised to their corresponding aldehydes and acids during aerobic culture, while aromatic aldehydes were both oxidised and reduced. However, the aromatic acids were not reduced, but were slowly metabolised. Dimer I was cleaved at the alkyl-phenyl linkage to give glycerol-2-guaiacyl ether in high yield. The identity of the latter was determined by mass spectrometry and proton nmr. The dimers 1-(3,4-dimethoxyphenyl)-2-(2-methoxyphenoxy)-propane-1,3-diol (II), 3,4-dimethoxy--(2,6-dimethoxy-4-carboxyphenoxy)-acetophenone (III) and 5-carboxy-7-methoxy-2-(4-hydroxy-3-methoxyphenyl)-3-methyl-2,3-dihydrobenzo-[b]-furan (IV) were not metabolised. It is concluded that the fungus resembles Fusarium in many of its metabolic properties, and could be considered as a potential lignin degrader.