• Korean Journal of Microbiology
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• v.48 no.1
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• pp.37-41
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• 2012

In this study, we screened and evaluated possibility of wood rot fungi as biocatalyst for biotransformation of sesquiterpenes. Screening were performed to select the most promising microorganisms with ability to biotransformation the substrate trans,trans-fanesol. Trans,trans-farnesol which is synthesized precursor of sesquiterpenes was used for resistance test on 19 of wood rot fungi. From the 19 tested wood rot fungi, 5 were selected by resistance test on different concentration of trans,trans-fanesol. Biotransformation was performed with selected wood rot fungi on liquid culture. The metabolites detected by GC-MS analysis were nerolidol for Laetiporus sulphureus var. miniatus (jungh) Imaz and eicosane for Coriolus versicolor (L.Fr) Prlar and isoborneol for Fomitopsis pinicola and isocyclocitral for Lampteromyces japonicas. As the results, wood rot fungi could be potential biocatalyst for biotransformation of sesquiterpenes.

• Journal of the Korean Applied Science and Technology
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• v.17 no.4
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• pp.213-225
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• 2000

Sophorolipids were biosynthesized using a strain of yeast, Torulopsis bombicola ATCC22214. It has been reported that this yeast gives the highest yields for the production of biosurfactant sophorolipids. Hence, this yeast was used in this study. One of the objectives of this study is to increase the yield of the sophorolipid synthesis. To meet this end, basic culture medium was formulated on the basis of literature research to-date. When this medium was used, the increase in yield from 15% to 150% was observed compared to using the media in the literature. To examine how the interfacial characteristics of sophorolipids change with substrate, glucose (the first carbon source) was maintained in the media and after being cultured for three days, the second carbon sources such as alkanes, vegetable oils, alcohols or organic acids were added. The whole broth was extracted twice with ethyl acetate and the extract was analyzed by thin layer chromatograhy(TLC). After qualitative analyses by TLC, surface tensions of sophorolipids were measured by the Wilhelmy plate method and critical micelle concentration(CMC) was determined using these surface tension data. Also, interfacial tensions were measured by the spinning drop method and emulsions of the three-component water/decane/sophorolipid system were tested. Sophorolipids were effective and efficient in terms of surface tension reduction and CMC, but they were ineffective as emulsifiers because emulsions were separated within 30 minutes.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.624-631
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• 2007

Thermostable protease is very effective to improve the industrial processes in many fields. Two thermostable extracellular proteases from the culture supernatant of the thermophilic fungus Chaetomium thermophilum were purified to homogeneity by tractional ammonium sulfate precipitation, ion-exchange chromatography on DEAE-Sepharose, and Phenyl-Sepharose hydrophobic interaction chromatography. By SDS-PAGE, the molecular mass of the two purified enzymes was estimated to be 33 kDa and 63 kDa, respectively. The two proteases were found to be inhibited by PMSF, but not by iodoacetamide and EDTA. The 33 kDa protease (PRO33) exhibited maximal activity at pH 10.0 and the 63kDa protease (PRO63) at pH5.0. The optimum temperature for the two proteases was $65^{\circ}C$. The PRO33 had a $K_m$ value of 6.6mM and a $V_{max}$ value of $10.31{\mu}mol/l/min$, and PRO63 l7.6mM and $9.08{\mu}mol/l/min$, with casein as substrate. They were thermostable at $60^{\circ}C$. The protease activity of PRO33 and PRO63 remained at 67.2% and 17.31%, respectively, after incubation at $70^{\circ}C$ for 1h. The thermal stability of the two enzymes was significantly enhanced by $Ca^{2+}$. The residual activity of PRO33 and PRO63 at $70^{\circ}C$ after 60min was approximately 88.59% and 39.2%, respectively, when kept in the buffer containing $Ca^{2+}$. These properties make them applicable for many biotechnological purposes.

• Journal of Microbiology and Biotechnology
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• v.25 no.1
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• pp.57-65
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• 2015

β-Glucosidase production by the white rot fungus Flammulina velutipes CFK 3111 was evaluated using different carbon and nitrogen sources under submerged fermentation. Maximal extracellular enzyme production was 1.6 U/ml, corresponding to a culture grown in sucrose 40 g/land asparagine 10 g/l. High production yield was also obtained with glucose 10 g/land asparagine 4 g/l medium (0.5 U/ml). Parameters affecting the enzyme activity were studied using p-nitrophenyl-β-_D-glucopyranoside as the substrate. Optimal activity was found at 50℃ and pHs 5.0 to 6.0. Under these conditions, β-glucosidase retained 25% of its initial activity after 12 h of incubation and exhibited a half-life of 5 h. The addition of MgCl₂, urea, and ethanol enhanced the β-glucosidase activity up to 47%, whereas FeCl₂, CuSO₄, Cd(NO₃)₂, and cetyltrimethylammonium bromide inflicted a strong inhibitory effect. Glucose and cellobiose also showed an inhibitory effect on the β-glucosidase activity in a concentration-dependent manner. The enzyme had an estimated molecular mass of 75 kDa. To the best of our knowledge, F. velutipes CFK 3111 β-glucosidase production is amongst the highest reported to date, in a basidiomycetous fungus.

• Journal of the Korean Wood Science and Technology
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• v.33 no.2 s.130
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• pp.65-71
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• 2005

This study was performed to understand the interaction between Ophiostomataceae and basidiomycetes fungi during cultures, and whether the basidiomycetes fungi inhibit the growth and decolorize dark pigments of blue staining fungi. The conjoint cultivation was studied on 2% malt extract agar. The ability of basidial cultures to decolorize dark pigments of ophiostomatoid fungi was the main characteristics estimated during this study. More than half of basidial cultures were characterized by deadlock interaction with blue staining fungi. In the dual cultures, where basidial partners were presented by Agaricus bisporus(64), Laetiporus sulphureus(L01/89), Trametes versicolor(09) and unknown fungus(02), antagonism was found at the phase of primary contact of colonies. Replacement interaction resulted usually in decreasing dark colour of substrate was observed for 11 basidial cultures that were belonging mainly to white-rot fungi. Among them Abortiporus biennis(123), Antrodiella hoehnelii(S28/91), Bjerkandera fumosa (137), and Gleophyllum odoratum(124) were characterized by the absence of deadlock-phase: they began to grow over dark colonies of their partners just after primary contact. Basidiomycetes did not affect strongly the pigments of Ceratocystis spp. and Leptographium sibirica isolates, but completely decolorized colonies of Ophiostoma ips and to a smaller degree Ophiostoma minus. Antrodiella hoehnelii(S28/91), Bjerkandera fumosa(137), Gleophyllum odoratum(124) and Trametes versicolor(B18/91) cultures were found to be the most active in decreasing dark color of blue staining fungi colonies. The cultures were recommended for further development as agents of biopulping of wood chips and bio-control of blue stain in woods.

• Korean Journal of Microbiology
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• v.14 no.2
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• pp.65-74
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• 1976

Atternaria sp. was isolated from soil and crude cellulases were prepared from wheat bran culture of the fungus. The activities of the crude enzyme were studied on five different subvstrates and some phsical properties were also examined, crude enzymes were purified by column chromatography on DEAE Sephadex and Sephadex, Isozymes were separated some of which were active specifically on DEAE cellulose and some were primarily active on cellulose and CM-cellulose. The optimal points of pH and temperature for the crude enzyme were varied depending on the substrates ; On cellulose they were at pH 6.0 and 40.deg.C, on CM-cellulose at pH's 4.0 and 6.0 and 60.deg.C, and on DEAW-cellulose at pH 5.0 and 50.deg.C. Two active fractions, F-1 and F-II on Na-CMC was used as substrate the Km values of crude enzyme, F-I and F-II were calculated to be $4{\times}10^{-5}$ , 1.1 * 10$^{-4}$ , and $1.25{\times}10^{-4}mN$ resepctively. The Ki value of $Cu^{++}$ for crude enzyme was$4{\times}^{-4}mN$ , while that of $Nm^{++}$ while in the same concentration of $Mn^{++}$ it reached to 91%. Some 57% activity of F-1 was inhibited in s mN $Cu^{++}$, whereas it was inhibited as much as 81% in the same concentration above the concentration of 0.3 mM with tis activity reaching up to 137% in 2 mM. On the other hand the F-11 was inhibited by the presence of M $n^{++}$ and some 67% activity was inhibited at 2mM.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.199-202
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• 2008

A white rot fungus Irpex lacteus produced lignin degrading enzymes, which showed degrading activity against various recalcitrant compounds. However, laccase, one of the lignin degrading enzymes, was too low to be assayed by spectrophotometry using o-tolidine as the chromogenic substrate in this fungus under various culture conditions. A laccase expression vector was constructed using a cDNA from Phlebia tremellosa with the constitutively expressed promoter of glyceraldehydes-3-phosphate dehydrogenase gene, and introduced into I. lacteus by the restriction enzyme mediated integration transformation through the protoplast-$CaCl_2$ procedure. Two transformants showed highly increased laccase activities at the early growth phase in the minimal liquid medium, and they not only degraded bisphenol A, a notorious endocrine disrupting chemical, but also removed the estrogenic activity effectively.

• Journal of Life Science
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• v.9 no.1
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• pp.26-34
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• 1999

A thermostable pullulanase has been isolated and purified from Thermus caldophilus GK-24 to a homogeneity by gel-filtration and ion-exchange chromatography. The specific activity of the purified enzyme was 431-fold increase from the crude culture broth with a recovery of 11.4%. The purified enzyme showed $M_{r}$ of 65 kDa on denaturated and natural conditions. The pI of the enzyme was 6.1 and Schiff staining was negative, suggesting that the enzyme is not a glycoprotein. The enzyme was most active at pH 5.5. The activity was maximal at $75^{\cire}C$ and stable up to $95^{\cire}C$ for 30 min at pH 5.5. The enzyme was stable to incubation from pH 3.5 to pH 8.0 at $4^{\cire}C$ for 24hr. The presence of pullulan protected the enzyme from heat inactivation, the extent depending upon the substrate concentration. The activity of the enzyme was simulated by $Mn^{2+}$ ion, }$Ni^{2+}$, $Ca^{2+}$, $Co^{2+}$ ions. The enzyme hydrolyzed the ${\alpha}$-1,6-linkages of amylopectin, glycogens, ${\alpha}$, ${\beta}$-limited dextrin, and pullulan. The enzyme caused the complete hydrolysis of pullulan to maltotriose and the activity was inhibited by $\alpha$, $\beta$, or $\gamma$-cyclodextrins. The $NH_{2}$-terminal amino acid sequence [(Ala-Pro-Gln-(Asp of Tyr)-Asn-Leu-Leu-Xaa-ILe-Gly-Ala(Ser)] was compared with known sequences of various sources and that was compared with known sequences of various sources and that was different from those of bacterial and plant enzymes, suggesting that the enzymes are structurally different.

• Food Science and Biotechnology
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• v.18 no.2
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• pp.500-505
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• 2009

The fibrinolytic enzyme, subtilisin D5, was purified from the culture supernatant of the isolated Bacillus amyloliquefaciens DJ-5. The molecular weight of subtilisin D5 was estimated to be 30 kDa. Subtilisin D5 was optimally active at pH 10.0 and $45^{\circ}C$. Subtilisin D5 had high degrading activity for the A$\alpha$-chain of human fibrinogen and hydrolyzed the $B{\beta}$-chain slowly, but did not affect the $\gamma$-chain, indicating that it is an $\alpha$-fibrinogenase. Subtilisin D5 was completely inhibited by phenylmethylsulfonyl fluoride, indicating that it belongs to the serine protease. The specific activity (F/C, fibrinolytic/caseinolytic activity) of subtilisin D5 was 2.37 and 3.52 times higher than those of subtilisin BPN' and Carlsberg, respectively. Subtilisin D5 exhibited high specificity for Meo-Suc-Arg-Pro-Tyr-pNA (S-2586), a synthetic chromogenic substrate for chymotrypsin. The first 15 amino acid residues of the N-terminal sequence of subtilisin D5 are AQSVPYGISQIKAPA; this sequence is identical to that of subtilisin NAT and subtilisin E.

• Korean Journal of Food Science and Technology
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• v.3 no.3
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• pp.185-188
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• 1971

This experiment was conducted to study on the productive conditions of cellulase by Aspergillus saitoi in the shaking culture medium. The results were as follows: 1. The production of enzyme required higher concentration of corn steep liquor than that of dextrin. 2. The concentration of 1.0% $NH_4H_2SO_4$ produced the enzyme excellently than 3.0%. 3. The cellulase was produced very slowly by adding $(NH_4)_2SO_4$, but the final concentration of the enzyme was higher than control. The production was suppressed by addition of $CaCO_3$. 4. The addition of $1.0{\sim}2.0%$ substrate caused an increase or stimulation in cellulase production.