• Journal of Life Science
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• v.20 no.5
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• pp.683-690
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• 2010

The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and $45^{\circ}C$, respectively, whereas those for the extracellular glucoamylase were 5.0 and $50^{\circ}C$. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

• Journal of Life Science
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• v.20 no.5
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• pp.662-669
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• 2010

Lipolytic enzyme-producing bacteria were isolated from edible oil mill effluents on tributyrin agar medium. The shake-flask-scale studies yielded a promising isolate and it was identified as Pseudomonas sp. An OME using various microbiological observations such as cultural, microscopic, and biochemical tests was undertaken and confirmed using PIBWIN bacterial identification software. Lipolytic enzyme production was screened with oils such as sunflower, caster, coconut, tributyrin, and olive. Amongst these, olive oil showed an increased lipase production 6.1 U/ml. In view of the highest lipolytic enzyme production with olive oil, further optimizations were carried out using olive oil as a carbon source. Lipolytic enzyme production was optimized by a conventional 'one variable at a time' approach and the significant factors were further analyzed statistically using response surface methodology (RSM). The effect of physical factors such as incubation time, temperature, initial medium pH, and nutritional factors such as concentration of olive oil and yeast extract were examined for lipase production. Lipolytic enzyme secretion was strongly affected by three variables (incubation time, concentration of yeast extract and olive oil). Therefore, the interaction of these three factors was further optimized using response surface methodology. The optimized conditions of lipase production using response surface methodology yielded a maximum of 9.62 U/ml with optimum conditions for incubation, yeast extract and olive oil concentrations were found to be 48 hr, 0.3 g. and 0.9 ml. respectively.

• Korean Journal of Microbiology
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• v.25 no.3
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• pp.221-228
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• 1987

The extracellular adenine deaminase from Nocardioides sp. J-275L was purified by the following techniques: ammonium sulfate fractionation, DEAE-Cellulose, DEAE-Sephadex A-50 column chromatography, and Sephacryl S-200 superfine gel filtration. The enzyme was partially purified about 3889.5-fold with about 5.2% yield by these procedures. The molecular weight of the enzyme was 39,000 by a calibrated Sephacryl S-200 superfine column chromatography. The enzyme was stable at pH 7.5 and up to $40^{\circ}C$. Glycerol was effective on the stabilization of the enzyme during storage. The optimum pH and temperature of the enzyme were around pH 7.5 and $40^{\circ}C$, respectively. The apparent Michaelis constant Km of the enzyme for adenine was $7.4\times 10^{-5}$M. The purine analogues, 6-chloropurine, 2,6-diaminopurine, 6-bromopurine, 4-aminopyrazolo [3.4-d]pyrimidine, and 8-azaadenine were substrates for the enzyme. 6-Dimethylaminopurine was a competitive inhibitor of the enzyme. The enzyme was inhibited by 1mM of $Cu^{2+}, Fe^{3+}, Pb^{2+}, Hg^{2+}$, and $Ag^{+}$, and 1mM of $\alpha$,$\alpha$'-dipyridyl, pentachlorophenol, and pCMB.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.223-229
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• 2006

To develop multifunctional microbial inoculant, an insluble phosphate-solubilizing bacterium with antifungal activity was isolated from plant rhizospheric soil. On the basis of its morphological, cultural and physiological characteristics and Biolog analysis, this bacterium was identified as Pseudomonas fluorescens RAF15. P. fluorescens RAF15 showed antifungal activities against phytopathogenic fungi Botrytis cinerea and Rhizoctonia solani. The optimal medium composition and cultural conditions for the solubilization of insoluble phosphate by P. fluorescens RAF15 were 1.5% of glucose, 0.005% of urea, 0.3% $MgCl_2{\cdot}6H_2\;0.01%\;of\;MgSO_4{\cdot}7H_2O\;0.01%,\;of\;CaCl_2{\cdot}2H_2O$, and 0.05% of NaCl along with initial pH 7.0 at $30^{\circ}C$. The soluble phosphate production under optimum condition was 863 mg/L after 5 days of cultivation. The solubilization of insoluble phosphates was associated with a drop in the pH of the culture medium. P. fluorescens RAF15 showed resistance against different environmental stresses like $10-35^{\circ}C$ temperature, 1-4% salt concentration and pH 2-11 range. The strain produced soluble phosphate to the culture broth with the concentrations of 971-1121 mg/L against $CaHPO_4$, 791-908 mg/L against $Ca_3(PO_4){_2}$, and 844 mg/L against hydroxyapatite, respectively. However, the strain produced soluble phosphate to the culture broth with the concentrations of 15 mg/L against $FePO_4$, and 5 mg/L against $AlPO_4$, respectively.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.339-345
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• 2009

Two lactic acid bacteria (LAB) with high ornithine-producing capacity were isolated from kimchi. Examination of the biochemical features using an API kit indicated that the strains belonged to the members of Weissella genus. They were gram positive, short rod-type bacteria, and able to grow anaerobically with $CO_2$ production. The isolates grew well on MRS broth at $25\sim37^{\circ}C$ and pH of 6.0~7.0. The optimum temperature and pH for growth are $30^{\circ}C$ and pH 6.5. The isolates fermented arabinose, ribose, xylose, glucose but not cellobiose, galactose, raffinose, or trehalsoe. The 16S rDNA sequences of isolates showed 99.6% and 99.7% homology with the Weissella koreensis S5623 16S rDNA (access no. AY035891). They were accordingly identified and named as Weissella koreensis OK1-4 and Weissella koreensis OK1-6, and could produce ornithine from MRS broth supplemented with 1% of arginine at a productivity of 27.01 and 31.41 mg/L/h, respectively. This is the first report on the production of ornithine by the genus Weissella isolated from kimchi.

• Korean Chemical Engineering Research
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• v.45 no.3
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• pp.258-263
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• 2007

Ammonia water was investigated as a new absorbent of the chemical absorption process for the removal of $CO_2$ in flue gas. The suitable range of ammonia water concentration and $CO_2$ loading ($mol\;CO_2/mol\;NH_3$) were decided in the point of view of $CO_2$ absorption capacity and $NH_4HCO_3$ precipitation. The absorption capacity of $CO_2$ and the precipitation of $NH_4HCO_3$ in liquid phase were calculated by the Pitzer model for electrolyte solution. The $CO_2$ absorption capacity of the ammonia water over $5\;molNH_3/kgH_2O$ was higher than that of conventional amine absorbent. The $CO_2$ loadings where precipitation occurred were decided at various absorbent concentrations. Theses values were higher than 0.5 in the concentration range of $5-14\;molNH_3/kgH_2O$ at 293, 313 K. The absorber for the removal of $CO_2$ in flue gas could be operated without $NH_4HCO_3$ precipitation by using high concentration of ammonia water below these $CO_2$ loading values. The optimum temperature of the ammonia water absorbent for removal of $CO_2$ in flue gas was 297-312 K depending on the concentration of ammonia water.

• Korean Chemical Engineering Research
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• v.56 no.2
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• pp.229-239
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• 2018

The purpose of this study was to effectively produce the biosugar from cell wall of lipid extracted microalgae (LEA) by using microwave-assisted pretreatment without enzymatic hydrolysis process. Response surface methodology (RSM) was applied to optimization of microwave-assisted pretreatment conditions for the production of biosugar based on enzyme-free process from LEA. Microwave power (198~702 W), extraction time (39~241 sec), and sulfuric acid (0~1.0 mol) were used as independent variables for central composite design (CCD) in order to predict optimum pretreatment conditions. It was noted that the pretreatment variables that affect the production of glucose (C6) and xylose (C5) significantly have been identified as the microwave power and extraction time. Additionally, the increase in microwave power and time had led to an increase in biosugar production. The superimposed contour plot for maximizing dependent variables showed the maximum C6 (hexose) and C5 (pentose) yields of 92.7 and 74.5% were estimated by the predicted model under pretreatment condition of 700 w, 185.7 sec, and 0.48 mol, and the yields of C6 and C5 were confirmed as 94.2 and 71.8% by experimental validation, respectively. This study showed that microwave-assisted pretreatment under low temperature below $100^{\circ}C$ with short pretreatment time was verified to be an effective enzyme free pretreatment process for the production of biosugar from LEA compared to conventional pretreatment methods.

• Korean Chemical Engineering Research
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• v.56 no.1
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• pp.119-124
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• 2018

The production of methyl N-phenyl carbamate by an oxidative carbonylation method of aniline and methanol is of great interest as an environmentally friendly process that can replace the monomer production process of a polymer produce using conventional phosgene. In this study, heterogeneous catalysts were prepared by using Y-zeolite, $SiO_2$, $Al_2O_3$ as support, and oxidative carbonylation continuous operation from aniline and methanol was attempted using the prepared heterogeneous catalyst. Batch reactor was used to determine the support, and various reaction conditions such as reaction temperature, reaction pressure, and effect of promoter were established using palladium catalyst. A reaction kinetics study was conducted under optimum reaction conditions. The basic data for carbamate process development were obtained by performing continuous operation for a long time under established reaction condition.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.6
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• pp.742-749
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• 2009

In order to establish the basic scientific data on the fermentation characteristics of chonggak kimchi (CGK), we studied with CGKs prepared at different seasons and stored at various temperatures ($5{\sim}23^{\circ}C$). Changes in pH and acidity of CGK fermented at different temperatures were typical. However, acidity of summer CGK stored under $15^{\circ}C$ did not reach 1.0% since the amount of reducing sugar in the summer radish was lower by over 1% than the ones from the other seasons. Growths for Leuconostoc and Lactobacillus spp. over 8.0 logCFU/mL were observed in CGKs fermented above $10^{\circ}C$. Changes in Max G, cutting force, of CGK from different seasons were apparent. Max Gs for spring and summer CGK were decreased immediately right after storage while Max Gs for winter CGKs were increased slowly during storage. According to the results of sensory evaluation, CGKs revealed the highest overall acceptability when acidity of kimchi reached $0.7{\pm}0.1%$ under the given fermentation condition. However, CGK prepared with radish from winter or summer season tasted better than the one made in the spring. But the edible periods for winter CGKs were longer, compared with summer CGKs fermented at the same temperature. In conclusion, CGK fermented at $10^{\circ}C$ in the winter season gives a better taste with longer edible duration than the other CGKs.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.1-7
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• 2002

Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.