Lenzites betulinus, known as gilled polypore belongs to Basidiomycota was isolated from fruiting body on broadleaf dead trees. It was found that the mycelia of white rot fungus Lenzites betulinus IUM 5468 produced ethanol from various sugars, including glucose, mannose, galactose, and cellobiose with a yield of 0.38, 0.26, 0.07, and 0.26 g of ethanol per gram of sugar consumed, respectively. This fungus relatively exhibited a good ethanol production from xylose at 0.26 g of ethanol per gram of sugar consumed. However, the ethanol conversion rate of arabinose was relatively low (at 0.07 g of ethanol per gram sugar). L. betulinus was capable of producing ethanol directly from rice straw and corn stalks at 0.22 g and 0.16 g of ethanol per gram of substrates, respectively, when this fungus was cultured in a basal medium containing 20 g/L rice straw or corn stalks. These results indicate that L. betulinus can produce ethanol efficiently from glucose, mannose, and cellobiose and produce ethanol very poorly from galactose and arabinose. Therefore, it is suggested that this fungus can ferment ethanol from various sugars and hydrolyze cellulosic materials to sugars and convert them to ethanol simultaneously.
Animals ; Arabinose ; Basidiomycota ; Biomass* ; Carbohydrates* ; Cellobiose ; Ethanol* ; Fruit ; Fungi* ; Galactose ; Gills ; Glucose ; Mannose ; Trees ; Xylose ; Zea mays

beta-Glucosidase, which hydrolyzes cellobiose into two glucoses, plays an important role in the process of saccharification of the lignocellulosic biomass. In this study, we optimized the activity of beta-glucosidase of brown-rot fungus Fomitopsis pinicola KCTC 6208 using the response surface methodology (RSM) with various concentrations of glucose, yeast extract and ascorbic acid, which are the most significant nutrients for activity of beta-glucosidase. The highest activity of beta-glucosidase was achieved 3.02% of glucose, 4.35% of yeast extract, and 7.41% ascorbic acid where ascorbic acid was most effective. The maximum activity of beta-glucosidase predicted by the RSM was 15.34 U/mg, which was similar to the experimental value 14.90 U/mg at the 16th day of incubation. This optimized activity of beta-glucosidase was 23.6 times higher than the preliminary activity value, 0.63 U/mg, and was also much higher than previous values reported in other fungi strains. Therefore, a simplified medium supplemented with a cheap vitamin source, such as ascorbic acid, could be a cost effective mean of increasing beta-glucosidase activity.
Ascorbic Acid ; beta-Glucosidase* ; Biomass ; Cellobiose ; Coriolaceae* ; Fungi* ; Glucose ; Vitamins ; Yeasts

A beta-glucosidase from Penicillium italicum was purified with a specific activity of 61.8 U/mg, using a chromatography system. The native form of the enzyme was an 88.5-kDa tetramer with a molecular mass of 354 kDa. Optimum activity was observed at pH 4.5 and 60degrees C, and the half-lives were 1,737, 330, 34, and 1 hr at 50, 55, 60, and 65degrees C, respectively. Its activity was inhibited by 47% by 5 mM Ni2+. The enzyme exhibited hydrolytic activity for p-nitrophenyl-beta-D-glucopyranoside (pNP-Glu), p-nitrophenyl-beta-D-cellobioside, p-nitrophenyl-beta-D-xyloside, and cellobiose, however, no activity was observed for p-nitrophenyl-beta-D-lactopyranoside, p-nitrophenyl-beta-D-galactopyranoside, carboxymetyl cellulose, xylan, and cellulose, indicating that the enzyme was a beta-glucosidase. The kcat/Km (s-1 mM-1) values for pNP-Glu and cellobiose were 15,770.4 mM and 6,361.4 mM, respectively. These values were the highest reported for beta-glucosidases. Non-competitive inhibition of the enzyme by both glucose (Ki = 8.9 mM) and glucono-delta-lactone (Ki = 11.3 mM) was observed when pNP-Glu was used as the substrate. This is the first report of non-competitive inhibition of beta-glucosidase by glucose and glucono-delta-lactone.
beta-Glucosidase ; Cellobiose ; Cellulases ; Cellulose ; Chromatography ; Citrus ; Fungi ; Glucose ; Glucosides ; Hydrogen-Ion Concentration ; Penicillium

During an investigation of fungi from an elm tree infested with bark beetles in Korea, one isolate, DUCC401, was isolated from elm wood. Based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer and 28S rDNA (large subunit) sequences, the isolate, DUCC401, was identified as Mariannaea samuelsii. Mycelia of the fungus grew faster on malt extract agar than on potato dextrose agar and oatmeal agar media. Temperature and pH for optimal growth of fungal mycelia were 25degrees C and pH 7.0, respectively. The fungus demonstrated the capacity to degrade cellobiose, starch, and xylan. This is the first report on isolation of Mariannaea samuelsii in Korea.
Agar ; Beetles ; Cellobiose ; DNA, Ribosomal ; Fungi ; Glucose ; Hydrogen-Ion Concentration ; Korea ; Solanum tuberosum ; Starch ; Ulmus ; Wood

A total of 106 Penicillium species were tested to examine their ability of degrading cellobiose, pectin and xylan. The activity of beta-glucosidase was generally strong in all the Penicillium species tested. P. citrinum, P. charlesii, P. manginii and P. aurantiacum showed the higher ability of producing beta-glucosidase than other tested species. Pectinase activity was detected in 24 Penicillium species. P. paracanescens, P. sizovae, P. sartoryi, P. chrysogenum, and P. claviforme showed strong pectinase activity. In xylanase assay, 84 Penicillium species showed activity. Strong xylanase activity was detected from P. megasporum, P. sartoryi, P. chrysogenum, P. glandicola, P. discolor, and P. coprophilum. Overall, most of the Penicillium species tested showed strong beta-glucosidase activity. The degree of pectinase and xylanase activity varied depending on Penicillium species.
beta-Glucosidase ; Cellobiose ; Penicillium* ; Polygalacturonase

Thirty seven species of Fusarium were evaluated for their ability of producing extracellular enzymes using chromogenic medium containing substrates such as starch, cellobiose, CM-cellulose, xylan, and pectin. Among the tested species Fusarium mesoamericanum, F. graminearum, F. asiaticum, and F. acuminatum showed high beta-glucosidase acitivity. Xylanase activity was strongly detected in F. proliferatum and F. oxysporum. Strong pectinase activity was also found in F. oxysporum and F. proliferatum. Amylase activity was apparent in F. oxysporum. No clear activity in cellulase was found from all the Fusarium species tested.
Amylases ; beta-Glucosidase ; Cellobiose ; Cellulase ; Fusarium* ; Polygalacturonase ; Starch

In the present study we first report in Korea the identification and characterization of Fusarium oxysporum isolated from rotten stems and roots of paprika (Capsicum annuum var. grossum) at Masan, Kyungsangnamdo in 2006. The fungal species produced white aerial mycelia accompanying with dark violet pigment on PDA. The optimal temperature and pH for the growth of the species was 25degrees C and pH 7, respectively. Microscopic observation of one of isolates of the species shows that its conidiophores are unbranched and monophialides, its microconidia have oval-ellipsoidal shape with no septate and are of 3.0~11 x 1.5~3.5 microm sizes, its macroconidia are of 15~20 x 2.0~3.5 microm sizes and have slightly curved or slender shape with 2~3 septate. The results of molecular analysis show that the ITS rDNA of F. oxysporum from paprika shares 100% sequence identity with that of known F. oxysporum isolates. The identified species proved it's pathogenicity by causing rotting symptom when it was inoculated on paprika fruits. The growth of F. oxysporum from paprika was suppressed on PDA by agrochemicals such as benomyl, tebuconazole and azoxystrobin. The identified species has the ability of producing extracelluar enzymes that degrade cellobiose and pectin.
Agrochemicals ; Benomyl ; Capsicum* ; Cellobiose ; DNA, Ribosomal ; Fruit ; Fusarium* ; Gyeongsangnam-do ; Hydrogen-Ion Concentration ; Korea* ; Viola ; Virulence

To evaluate which dye is effective in a plate assay for detecting extracellular cellulase activity produced by fungi, four chromogenic dyes including remazol brilliant blue, phenol red, congo red, and tryphan blue, were compared using chromagenic media. For the comparison, 19 fungal species belonging to three phyla, ascomycota, basidiomycota, and zygomycota were inoculated onto yeast nitrogen-based media containing different carbon substrates such as cellulose (carboxylmethyl and avicel types) and cellobiose labeled with each of the four dyes. Overall, the formation of clear zone on agar media resulting from the degradation of the substrates by the enzymes secreted from the test fungi was most apparent with media containing congo red. The detection frequency of cellulase activity was also most high on congo red-supplemented media. The results of this study showed that congo red is better dye than other three dyes in a plate assay for fungal enzyme detection.
Agar ; Ascomycota ; Basidiomycota ; beta-Glucosidase ; Carbon ; Cellobiose ; Cellulase ; Cellulases* ; Cellulose ; Coloring Agents* ; Congo ; Congo Red ; Fungi* ; Phenolsulfonphthalein ; Yeasts

Cellulosic material is the most abundant renewable carbon source in the world. Cellulose may be hydrolyzed using cellulase to produce glucose, which can be used for production of ethanol, organic acids, and other chemicals. Cellulase is a complex enzyme containing endoglucanase (EC 3.2.1.4), exoglucanase (EC 3.2.1.91) and cellobiase (EC 3.2.1.21). The hydrolysis of natural cellulose to glucose depends on the synergism of these three components. The mostly used cellulase produced by Trichoderma reesei has high activity of endoglucanase and exoglucanase, but the activity of cellobiase is relatively low. Therefore, improving the activity of cellobiase in cellulase reaction system is the key to enhance the sacchrification yield of cellulosic resources. Aspergillus niger LORRE 012 was a high productivity strain for cellobiase production. It was found that the spores of this strain were rich in cellobiase. In this work, the cellobiase was immobilized efficiently by simply entrapping the spores into calcium alginate gels instead of immobilizing the pure cellobiase proteins. The immobilized cellobiase was quite stable, and its half-life was 38 days under pH 4.8, 50 degrees C. The thermal stability of the immobilized cellobiase was improved, and it was stable below 70 degrees C. The suitable pH range of the immobilized cellobiase was pH 3.0 - 5.0, with the optimal pH value 4.8. The Km and Vmax value of the immobilized cellobiase were 6.01 mmol/L and 7.06 mmol/min x L, respectively. In repeated batch hydrolysis processes, 50 mL of substrate (10 g/L cellobiose) and 10 mL of immobilized beads containing cellobiase were added into a 150 ml flask. After reacting at pH 4.8, 50 degrees C for several hours, the hydrolysate was harvested for assay, and the immobilized beads were used for the next batch hydrolysis with the fresh substrate. This process was repeated, and the yield of enzymatic hydrolysis kept higher than 97% during 10 batches. The continuous hydrolysis process was carried out in a column reactor (inside diameter 2.8 cm, inside height 40 cm) packed with the immobilized beads. Using 10 g/L cellobiose as substrate, the hydrolysis yield reached 98% under 0.4 h (-1) dilution rate and pH 4.8, 50 degrees C. After corncob was treated by 1% dilute acid, the cellulosic residue (100 g/L) was used as substrate, and hydrolyzed by the cellulase (15 IFPU/g substrate) from Trichoderma reesei, at pH 4.8, 50 degrees C for 48 h. The concentration of reducing sugar in the hydrolysate was only 48.50 g/L (hydrolysis yield 69.5%). When the hydrolysate was further treated by the immobilized cellobiase, the cellobiose was hydrolyzed into glucose, and the feedback inhibition caused by the cellobiose accumulation disappeared sharply. By the synergism of immobilized cellobiase and the cellulase from T. reesei left in the hydrolysate, other oligosaccharides were mostly converted to monosaccharides. At 48 h, the reducing sugar concentration was increased to 58.78 g/L, the hydrolysis yield of the corncob residue was improved to 84.2%, and the ratio of the glucose in the total reducing sugar was increased from 53.6% to 89.5%. The reducing sugars converted from corncob could be used further in the fermentation of valuable industrial products. This research results were meaningful in the conversion and utilization of renewable biomass.
Aspergillus niger ; enzymology ; Biotechnology ; Cellobiose ; metabolism ; Enzyme Stability ; Hydrogen-Ion Concentration ; Kinetics ; Temperature ; beta-Glucosidase ; chemistry ; metabolism

BACKGROUND: Vibrio(V.) vulnificus is a halophilic, gram-negative bacillus that causes a fatal sepsis in patients with underlying chronic disease such as liver cirrhosis and alcoholic abuse. Because V. vulnificus infection has a fulminant course and high mortality rate, early recognition and rapid diagnosis with prompt therapy are necessary to improve survival rate. OBJECTIVE: The purpose of this study was to develop a new selective medium for rapid identification of V. vulnificus through color change of medium according to pH from patients suspected of having V. vulnificus sepsis. METHODS: Rapid isolation and identification of V. vulnificus can be possible by modifying the component of PNC(5% peptone, 1% NaCl, and 0.08% cellobiose [pH 8.0]) broth medium. From this PNC broth, a basal broth(5% peptone+1% NaCl+cellobiose) was prepared and used to evaluate additional medium supplements(cellobiose concentration [0.08, 0.2, 0.1%], pH [6.8, 7.5, 8.0] and pH indicator dye [bromthymol blue, thymol blue, phenol red, bromcresol purple, crystal violet, cresol red, and neutral red]). To examine the rapid identification and selectivity of this basal medium according to various conditions, V. vulnificus was tested by using saline and normal human blood containing these bacteria(1, 000 bacteria/ml), respectively at 37degrees C. A positive reaction(V. vulnificus growth) appeared as color change. The selectivity and identification capacity of this new broth was tested by using other 6 Vibrio species and 14 strains of other bacteria. RESULTS: Color change appeared only in the medium including bromthymol blue and thymol blue as a pH indicator dye. It was called the basal medium containing blue dyes as PNCB(peptone, NaCl, cellobiose and blue dye) medium. It took an average time of 4.8hr for becoming aware of yellow color change in PNCB broth after cultivating with saline mixed with V. vulnificus and 6hr in PNCB broth after cultivating with blood mixed with V. vulnificus. One Vibrio species and another 3 bacteria produced color change. So we confirmed that the final composition and pH of PNCB broth medium was 5% peptone, 1% NaCl, 0.2% cellobiose, 0.0004% bromthymol blue and 0.0004% thymol blue [pH 7.5] CONCLUSIONS: PNCB broth could be used as a selective and differential medium for rapid isolation and identification of V. vulnificus in patients with V. vulnificus sepsis.
Alcoholics ; Bacillus ; Bacteria ; Bromcresol Purple ; Bromthymol Blue ; Cellobiose ; Chronic Disease ; Coloring Agents ; Diagnosis ; Gentian Violet ; Humans ; Hydrogen-Ion Concentration ; Liver Cirrhosis ; Mortality ; Peptones ; Phenolsulfonphthalein ; Sepsis* ; Survival Rate ; Thymol ; Vibrio vulnificus* ; Vibrio*