Screening, purification, and characterization of an extracellular lipase from Aureobasidium pullulans isolated from stuffed buns steamers / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

An extracellular lipase from Aureobasidium pullulans was obtained and purified with a specific activity of 17.7 U/mg of protein using ultrafiltration and a DEAE-Sepharose Fast Flow column. Characterization of the lipase indicated that it is a novel finding from the species A. pullulans. The molecular weight of the lipase was 39.5 kDa, determined by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited its optimum activity at 40 °C and pH of 7. It also showed a remarkable stability in some organic solutions (30%, v/v) including n-propanol, isopropanol, dimethyl sulfoxide (DMSO), and hexane. The catalytic activity of the lipase was enhanced by Ca2+ and was slightly inhibited by Mn2+ and Zn2+ at a concentration of 10 mmol/L. The lipase was activated by the anionic surfactant SDS and the non-ionic surfactants Tween 20, Tween 80, and Triton X-100, but it was drastically inhibited by the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). Furthermore, the lipase was able to hydrolyze a wide variety of edible oils, such as peanut oil, corn oil, sunflower seed oil, sesame oil, and olive oil. Our study indicated that the lipase we obtained is a potential biocatalyst for industrial use.

Screening, purification, and characterization of an extracellular lipase from Aureobasidium pullulans isolated from stuffed buns steamers / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

An extracellular lipase from Aureobasidium pullulans was obtained and purified with a specific activity of 17.7 U/mg of protein using ultrafiltration and a DEAE-Sepharose Fast Flow column. Characterization of the lipase indicated that it is a novel finding from the species A. pullulans. The molecular weight of the lipase was 39.5 kDa, determined by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited its optimum activity at 40 °C and pH of 7. It also showed a remarkable stability in some organic solutions (30%, v/v) including n-propanol, isopropanol, dimethyl sulfoxide (DMSO), and hexane. The catalytic activity of the lipase was enhanced by Ca2+ and was slightly inhibited by Mn2+ and Zn2+ at a concentration of 10 mmol/L. The lipase was activated by the anionic surfactant SDS and the non-ionic surfactants Tween 20, Tween 80, and Triton X-100, but it was drastically inhibited by the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). Furthermore, the lipase was able to hydrolyze a wide variety of edible oils, such as peanut oil, corn oil, sunflower seed oil, sesame oil, and olive oil. Our study indicated that the lipase we obtained is a potential biocatalyst for industrial use.

Efficient production of pullulan by Aureobasidium pullulans grown on mixtures of potato starch hydrolysate and sucrose

Abstract Pullulan is a natural exopolysaccharide with many useful characteristics. However, pullulan is more costly than other exopolysaccharides, which limits its effective application. The purpose of this study was to adopt a novel mixed-sugar strategy for maximizing pullulan production, mainly using potato starch hydrolysate as a low-cost substrate for liquid-state fermentation by Aureobasidium pullulans. Based on fermentation kinetics evaluation of pullulan production by A. pullulans 201253, the pullulan production rate of A. pullulans with mixtures of potato starch hydrolysate and sucrose (potato starch hydrolysate:sucrose = 80:20) was 0.212 h−1, which was significantly higher than those of potato starch hydrolysate alone (0.146 h−1) and mixtures of potato starch hydrolysate, glucose, and fructose (potato starch hydrolysate:glucose:fructose = 80:10:10, 0.166 h−1) with 100 g L−1 total carbon source. The results suggest that mixtures of potato starch hydrolysate and sucrose could promote pullulan synthesis and possibly that a small amount of sucrose stimulated the enzyme responsible for pullulan synthesis and promoted effective potato starch hydrolysate conversion effectively. Thus, mixed sugars in potato starch hydrolysate and sucrose fermentation might be a promising alternative for the economical production of pullulan.

Characterization of pectinase activity for enology from yeasts occurring in Argentine Bonarda grape

Pectinolytic enzymes are greatly important in winemaking due to their ability to degrade pectic polymers from grape, contributing to enhance process efficiency and wine quality. This study aimed to analyze the occurrence of pectinolytic yeasts during spontaneous fermentation of Argentine Bonarda grape, to select yeasts that produce extracellular pectinases and to characterize their pectinolytic activity under wine-like conditions. Isolated yeasts were grouped using PCR-DGGE and identified by partial sequencing of 26S rRNA gene. Isolates comprised 7 genera, with Aureobasidium pullulans as the most predominant pectinolytic species, followed by Rhodotorula dairenensis and Cryptococcus saitoi. No pectinolytic activity was detected among ascomycetous yeasts isolated on grapes and during fermentation, suggesting a low occurrence of pectinolytic yeast species in wine fermentation ecosystem. This is the first study reporting R. dairenensis and Cr. saitoi species with pectinolytic activity. R. dairenensis GM-15 produced pectinases that proved to be highly active at grape pH, at 12 °C, and under ethanol and SO₂ concentrations usually found in vinifications (pectinase activity around 1.1 U/mL). This strain also produced cellulase activity at 12 °C and pH 3.5, but did not produce β-glucosidase activity under these conditions. The strain showed encouraging enological properties for its potential use in low-temperature winemaking.

Rsearch progress on breeding of pullulan high-yield strain without melanin / 中国生化药物杂志

Pullulan is a linear glucosic polysaccharide produced by the polymorphic fungus Aureobasidium Pullulans, which has long been applied for various applications in medical and food industry due to its security, stability and low adhesive ability.At present, the two problems in restricting pullulan industrial production are the low polysaccharide production and melanin secreted which is hard to erase completely, giving the following process some problem.As a starting point, this review article collects and analyzes the progress on the breeding of pullulan high-yield strain without melanin in recent years, in order to find more efficient strains breeding methods, laying a foundation for further breeding of pullulan high-yield strain without melanin.

Aureobasidium pullulans infection in a patient with chronic lymphocytic leukemia

Saprophytic fungi are being increasingly recognized as etiologic agents of mycoses in immunosuppressed patients. We report a case of subcutaneous infiltration by Aureobasidium pullulans, likely due to traumatic inoculation, in a neutropenic patient during chemotherapy for chronic lymphocytic leukemia. The patient was treated with amphotericin B deoxycholate but was subsequently switched to itraconazole, which improved the lesion. This case highlights the importance of considering unusual fungal infections in critically ill patients such as those who are immunosuppressed due to chemotherapy. Diagnostic techniques and effective antifungal therapy have improved the prognosis of these cases.

Production of pullulan by a thermotolerant Aureobasidium pullulans strain in non-stirred fed batch fermentation process

Total 95 isolates of Aureobasidium pullulans were isolated from different flowers and leaves samples, out of which 11 thermotolerant strains produced pullulan. One thermotolerant non-melanin pullulan producing strain, designated as RG-5, produced highest pullulan (37.1±1.0 g/l) at 42ºC, pH 5.5 in 48h of incubation with 3% sucrose and 0.5% ammonium sulphate in a non-stirred fed batch fermentor of 6 liters capacity. The two liters of initial volume of fermentation medium was further fed with the 2 liters in two successive batches at 5 h interval into the fermentor. The sterile air was supplied only for 10h at the rate of 0.5 vvm.

Properties of alkaline protease genetically engineered on cell surface of the yeast Yarrowia lipolytica.

ALP2 gene encoding alkaline protease cloned from Aureobasidium pullulans HN2-3 was ligated into the surface display plasmid and expressed in the cells of the yeast Yarrowia lipolytica. The expressed alkaline protease was immobilized on the yeast cells. The activity of the immobilized enzyme with 6 His tag was found to be significantly higher than that of without 6 His tag. The immobilized enzyme showed lower optimal temperature and a lower affinity for azocasein than the free enzyme purified from A. pullulans HN2-3. The thermal stability of the immobilized enzyme enhanced and the pH stability decreased, compared to that of the free enzyme.

Aureobasidium Pullulans Sepsis Developing in Patient, Who Received Kidney Transplantation in China / 대한이식학회지

Aureobasidium Pullulans (AP) is a fungus known as a "black fungus" characterized by production of melanin pigment. It is infected mainly by subcutaneous pathway, but causes a very rare disease in human. There was a very few report of AP infection in human all around the world and no report in Korea, yet. We reported the first Korean case of AP sepsis patient who received the second kidney transplantation in China. The patient was a 61-year old male who underwent the first kidney transplantation at Severance Hospital in July 1992 and had experienced chronic renal graft dysfunction for the last several years. He went to China and underwent the 2nd kidney transplantation from acute brain-injury donor on December 31, 2002. He discharged and came back to Korea at POD #14 and admitted to our department. At admission, there was no specific symptom or sign of infection and the function of allograft kidney was good with serum creatinine of 1.2 mg/dL. He was on tacrolimus 4 mg bid, deltacortef 10mg bid and MMF 1.0 gm bid. During the hospitalization, the dosage of tacrolimus was controlled by adjusting serum tacrolimus level around 10~15 ng/mL, and reduced the dosage of deltacortef to 5 mg bid and MMF to 500 mg bid. Since Zenapax was already administered during the hospitalization in China, we added additional injection of Zenapax twice with 2 weeks interval. On POD #22, he developed skin rash and edema compatible to cellulites on the intravenous puncture site of left upper arm during his hospital stay in China. MMF was stopped and broad spectrum antibiotic therapy was started immediately. On POD #23, he developed acute myocardiac infarction and he undertook PTCA with arterial stent insertion. He was transported to intensive care unit due to acute respiratory failure on POD #27, and the left arm color was changed to black on POD #30. The empirical intravenous amphotericin therapy was started at POD #35, but the patientdied due to multiple organ failure caused by fungal sepsis. After his death, we received positive culture report of AP from his blood and skin lesion specimens collected on POD #29 and 35.

A Case of Onychomycosis due to Aureobasidium pullulans after Trauma / 대한의진균학회지

Aureobasidium pullulans is classified among dematiaceous fungi, characterized by the production of melanin pigments. It is widely distributed throughout the environment. It has occasionally been infected on the skin and nails of humans. We report a case of onychomycosis due to Aureobasidium pullulans developed in an 49-year-old female after trauma.

THE EFFECTS OF A CONTROLLED pH ENVIRONMENT ON POLYSACCHARIDE SYNTHESIS BY AUREOBASIDIUM PULLULANS / 微生物学通报

The effect of pH on the extracellularpolysaccharide synthesis by Aureobasidium pullulans was studied by addding CaCO 3and HCl.Cultivated in P2 liquid medium for 24h,pH dropped to 3.6 because of strongly producing acid.Under this low pH environment,further fermentation for 120h,only 5.9g/L of polysacharide was obtained.When grown in MP2 medium contaning0.5% CaCO 3,the pH was kept above 5.0 during 144 hours,production of polysaccharide increased to 31g/L. The detailed information of effects of controlled pH on polysaccharide production showed an optimal pH value 5.0 must be maintained through the fermentative period.