• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.740-746
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• 2021

Efficient cellulolytic enzyme production is important for the development of lignocellulose-degrading enzyme mixtures. However, purification of cellulases from their native hosts is time- and labor-consuming. In this study, a constitutive expression system was developed in Penicillium oxalicum for the secreted production of proteins. Using a constitutive polyubiquitin gene promoter and cultivating with glucose as the sole carbon source, nine cellulolytic enzymes of different origins with relatively high purity were produced within 48 h. When supplemented to a commercial cellulase preparation, cellobiohydrolase I from P. funiculosum and cellobiohydrolase II from Talaromyces verruculosus showed remarkable enhancing effects on the hydrolysis of steam-exploded corn stover. Additionally, a synergistic effect was observed for these two cellobiohydrolases during the hydrolysis. Taken together, the constitutive expression system provides a convenient tool for the production of cellulolytic enzymes, which is expected to be useful in the development of highly efficient lignocellulose-degrading enzyme mixtures.

• Microbiology and Biotechnology Letters
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• v.10 no.4
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• pp.267-273
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• 1982

Among 12 kinds of ginsenosides in ginseng saponin, ginsenoside-Rb$_1$was contained the most abundantly. But ginsenoside-Rd which is similar to ginsenoside-Rb$_1$in structure, was known to be superior to ginsenoside-Rb$_1$pharmaceutically. In order to convert ginsenoside-Rb$_1$into ginsenoside-Rd by microbial enzyme treatment, a Rhizopus sp. was selected among various strais of molds found in rotten ginseng roots. Enzyme was prepared from the extract of wheat bran koji culture by ammonium sulfate precipitation (1.0 sat'd) and succeeding ammonium sulfate fractionation method (0.6-0.9 sat'd). For the purpose of use as substrate, saponins were purified by the several purification steps from alcohol extract of red ginseng roots. We obtained the total saponin which was composed of 36.5% of ginsenoside Rb$_1$, 12.2% of ginsenoside-Rd and other ginsenosides. For increase of ginsenoside-Rb$_1$ component ratio, we also obtained further purified ginsenoside-Rb group saponin containing 54.5% of ginsenoside-Rb$_1$, 1.1% of ginsenoside- Rd and other ginsenosides from purified the total saponin. In the enzymatic reaction system including the total saponin or the ginsenoside-Rb group saponin, we confirmed the specific conversion of ginsenoside-Rb$_1$to ginsenoside-Rd proportionally and no change of any other ginsenoside patterns by thin layer chromatography and high performance liquid chromatography.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.46 no.4
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• pp.349-360
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• 2020

Bio-conversion manufacturing technology has been developed to produce ginsenoside Rd which is increasingly in demand as a cosmetic material due to various possibilities related to improving skin function. In order to convert ginsenoside Rb1 which is contained in red ginseng saponin (RGS) into Rd, several commercial enzymes were tested. Viscoflow MG was found to be the most efficient. In order to optimize the conversion of RGS to ginsenoside Rd by enzymatic transition was carried out using response surface methodology (RSM) based on Box-Behnken design (BBD). The main independent variables were RGS concentration, enzyme concentration, and reaction time. Conversion of ginsenoside Rd was performed under 17 conditions selected according to BBD model and optimization conditions were analyzed. The concentration of the converted ginsenoside Rd ranged from 0.3113 g/L to 0.5277 g/L, and the highest production volume was obtained under condition of reacting 2% RGS and 1.25% enzyme for 13.5 hours. Consequently, RGS concentration, enzyme concentration which is 0.05 less than p-value and among the interactions between the independent variables, the interaction between enzyme concentration and reaction time was confirmed to be the most influential.

• Journal of fish pathology
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• v.6 no.2
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• pp.133-142
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• 1993

Vitamin B6(pyridoxine, pyridoxamine. and pyridoxal) is a dietary requirement in relatively small quantities for growth, health, and function in animals and fish. The metabolically active B6 is pyridoxal-5-phosphate(PLP). It does function as a coenzyme in number of enzymes(PLP-dependent enzymes) in which amino acids are metabolized, including decarboxylases, aminotransferases, sulfhydrases, tryptophanase, and hydroxylases. Vitamin B6 requirement is higher for fish because fish are fed much higher protein diet than land animals. B6 is also involved in metabolism of carbohydrates and lipids and essential for the synthesis of heme and serotonin. Deficiency signs in fish develop quickly, in cluding nervous disorders, convulsions, poor swimming coordination, skin lesions, edema, exophthalmos, and tetany. The conversion of vitamin B6 to metabolically active form(PLP) is catalyzed by pyridoxal kinase and pridoxine(pyridoxamine) oxidase. In this review, we summarized in detail the enzymatic studies on vitamin B6 metabolism and about the mechanisms and properties of a PLP-dependent enzyme.

• Journal of the Korean Wood Science and Technology
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• v.43 no.3
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• pp.344-354
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• 2015

To investigate the effects of ionic liquid pretreatment on biomass, giant miscanthus was treated with 1-ethyl-3-methylimidazolium acetate ([Emim][OAc]) and 1-butyl-3-methylimidazolium acetate ([Bmim][OAc]) at three temperature conditions ($90^{\circ}C$, $110^{\circ}C$, and $130^{\circ}C$). As temperature condition increased, yield of the cellulose-rich product (CP) was reduced from 87.2% to 67.6%, while yield of the ionic liquid lignin (ILL) increased from 2.2% to 9.9%. Compared to the ILL, CP had lower carbon contents and higher oxygen contents. Enzymatic hydrolysis of CPs showed that conversion ratio of CP treated with [Emim][OAc] at $110^{\circ}C$ was 56.7%, the highest digestibility. Thermogravimetric analysis indicated that the maximum degradation rate decreased as temperature condition increased. In addition, maximum degradation temperature of ILL treated with [Emim][OAc] ranged from 274 to $279^{\circ}C$ which was lower than that of ILL treated with [Bmim][OAc]. Analytical date for ${\beta}$-O-4 linkage frequency in the ILL revealed that ${\beta}$-O-4 linkage frequency in the ILL decreased as the temperature rose. Furthermore, the highest S/G ratio of the ILL was determined to ca. 1.2 obtained from [Bmim][OAc] treatment at $110^{\circ}C$.

• Journal of the Korean Wood Science and Technology
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• v.39 no.1
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• pp.95-103
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• 2011

We have investigated pretreatment of waste wood using milling refinery combined with poping method, which can save energy for pretreatment and enzyme loading for enzymatic hydrolysis. The chemical analysis of holocellulose of non and popping treated waste wood showed 65.9% and 58.8%, and the lignin, organic extracts and ash were increased by 3%, 4% and 0.7% after pretreatment, respectively. The reducing sugar yields of pretreated waste wood were increased four times more than non-pretreated one and the synergistic effect of cellulase and xylanase were evaluated compare with individual enzyme treatment. Especially, enzyme cocktail (cellulase 50 U and xylanase 50 U) treatment was very efficient in 1% substrate (50 mg). Also, glucose and xylose conversion rate of pretreated waste wood by GC analysis were 45.9% and 38.7%, respectively.

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.44-49
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• 1996

By using the ${\beta}$-tyrosinase of Citrobacter freundii KCT2006, which was cloned and overexpressed in Escherichia coli, 3,4-dihydroxy phenyl-L-alanine (L-DOPA) was synthesized efficiently from pyrocatechol, sodium pyruvate, and ammonium acetate. Optimal temperature and pH for the reaction were determined to be about 18$^{\circ}C$ and 8.5, respectively. The effects of substrate concentrations were also examined at different concentrations of ammonium acetate, sodium pyruvate, and pyrocatechol. Ammoniumacetate and sodium pyruvate increased the reaction rate until the concentrations reached to 300mM and 50mM, respectively. Although pyrocatechol showed the optimal concentration at 20mM, it was controlled between 20mM and 50mM to avoid the depletion of substrate during the enzymatic synthesis. Meanwhile the synthetic rate was improved about 20% when ethanol was included in the reaction solution. Based on above results, a reaction medium for the productin of L-DOPA was prepared and incubated with 1 unit/ml of ${\beta}$-tyrosinase. Pyrocatechol and sodium pyruvate was added to the reaction solutin intermittently to avoid the substrate depletion during the enzymatic reaction. After 24 hour of reaction, 31.6g/l of L-DOPA was accumulated in the reaction solution as soluble and precipitated ones and the conversion yield was about 85.2%.

• Microbiology and Biotechnology Letters
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• v.26 no.1
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• pp.15-22
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• 1998

The dnrF gene, responsible for conversion of aklavinone to $\varepsilon$-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin gene cluster of Streptomyces peucetius subsp. caesius ATCC 27952, close to drrAB, one of the anthracycline resistance genes. To characterize the enzymatic properties of the aklavinone 11-hydroxylase, the dnrF gene was overexpressed in Escherchia coli. The pET-22(+) plasmid which has the T7 promoter under the control of lacUV5 gene was used for the overexpression of the dnrF gene, and the recombinant plasmid pET213 that contains the dnrF gene linked to the T7 promoter of pET-22b(+) was introduced into the E. coli BL2l. When the expression of the dnrF gene was induced by IPTG at the final concentration of 1 mM, the induced protein could be detected in SDS-PAGE only in insoluble precipitate. The insoluble protein was electroeluted from the gel and used for the preparation of antiserum in mice. Various culture conditions were tested to maximize the expression of the aklavinone 11-hydroxylase in soluble form. The enzymatic activity was checked by the bioconversion experiment, and the protein was confirmed by the SDS-PAGE and the Western blot analysis. From the analysis of the data, it was concluded that the culture induced with IPTG at the final concentration of 0.02 mM at 37$^{\circ}C$ yielded the best productivity of active form of enzyme.

• Applied Biological Chemistry
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• v.14 no.1
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• pp.19-27
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• 1971

This experiment was conducted as a preliminary test to invent the quick ripening process for soysauces. The bran koji extract of 2 kinds of mold (Asp. niger and Asp. sojae) was used as enzyme sources in order to improve the rate of enzymatic conversion of raw materials and shorten the ripening period of soysauces. Soybean koji (inoculated Asp. sojae) and steamed soybean were enzymatically decomposed for 8 hours at $40^{\circ}C$, pH 4-6 with stirring at a rate of 1500 rpm. After the enzymatic treatment, soysauces were fermented by 4 kinds of processes with the above-mentioned salt tolerant microorganisms inoculated. After 4 weeks, sensory evolution for these soysauces products and analysis of chemical components were carried out. The results obtained were summarized as follows. 1. Soysauces prepared from decomposed koji by Asp. niger enzyme complex showed higher extractables, such as total nitrogen, amino nitrogen, pure extract and reducing sugar, than any other procedure. Extracted amounts of each components of soysauces prepared from this process during 1 month were similar to those of soysauces made from general procedure (control) for 5 to 6 months. 2. According to sensory evaluation, soysauce prepared from decomposed koji also showed best taste and were similar to that of soysauces prepared by general method for 6 months.

• Korean Journal of Food Science and Technology
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• v.26 no.5
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• pp.567-572
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• 1994

Diglyceride was prepared by reaction of hydrogenated beef tallow and glycerol (GL) in the presence of a Pseudomonas lipase. Both substrates were mixed at the ratio of GL/Triglyceride of 0.5 which is the stoichiometric molar ratio for the complete conversion of triglyceride (TG) to diglyceride (DG). DG can be obtained by solid phase-glycerolysis of hydrogenated beef tallow without use of organic solvents or emulsifiers by careful control of reaction temperature. Optimized reaction temperature condition was as follows: An initial incubation at$60^{\circ}C$ for 2h followed by the first temperature shift down to $55^{\circ}C$ for 4h, and then the second shift down to $50^{\circ}C$ for up to 3 days. There was a large decrease in the content of TG during the first $60^{\circ}C$ incubation for 2h. Even a prolonged incubation at $60^{\circ}C$ could not make a change of the composition of the reaction mixture at liquid state. By controlling the temperature lower than $60^{\circ}C$, reaction mixtures were solidified. The reaction temperature at $50^{\circ}C$ below the melting temperature of hydrogenaed beef tallow gave an 71% optimum yield of DG after 72h enzymatic glycerolysis and about 73% of total DG was 1,3-DG.