• Applied Biological Chemistry
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• v.42 no.1
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• pp.1-5
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• 1999

For effective utilization of apple pomace, protopectinase(PPase) produced from Bacillus subtilis IFO 12113 was used for pectin extraction from apple pomace. Optimal conditions for enzyme treatment were found at $60^{\circ}$, pH 7.8, 48 hours with 20 : 1 ratio of substrate to enzyme. The yield of extracted pectin from water-alcohol insoluble pectin by enzyme at optimal condition was 34.3%. The purity and methoxyl content of extracted pectin by PPase at optimal condition were measured as 52.9% and 2.75%, respectively. Intrinsic viscosity and average molecular weight of extracted pectin by enzymatic method were 0.178 ml/g and $4.9{\times}10^3$, respectively.

• Journal of The Korean Society of Agricultural Engineers
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• v.60 no.1
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• pp.1-10
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• 2018

This study analyzed pore water pressure, seepage and leakage quantity, height of seepage and critical hydraulic gradient in order to suggest the seepage characteristics of agricultural reservoir embankment considering filter interval. The seepage characteristics of a deteriorated reservoir embankments were conducted according to the horizontal filter intervals range using three- dimensional finite element analysis. The wider the horizontal filter interval, the higher the pore water pressure increased, and the pore water pressure ratio in the center of the core has a greater effect than the base part. The seepage and leakage quantity appeared largely in the two-dimensional analysis conditions (case 1), where the filter was constructed totally in the longitudinal direction of the embankment, the wider the horizontal filter interval was gradually reduced. The reasonable filter intervals to yield efficient seepage characteristics were within 30 m for the pore water pressure of the core and the height of the seepage line. The stability of the filter installation was able to evaluate the stability of the piping by the critical hydraulic gradient method. The deteriorated reservoir with no filters or decreased functionality can significantly reduce the possibility of piping by simply installing a filter on the downstream slope. In the future, the deteriorated reservoir embankment should be checked for the reservoir remodeling because the core and filter functions have been lost or decreased significantly. In the case of a new installation, the seepage characteristic behavior due to the core and filter changes should be applied to the field after obtaining a reasonable horizontal filter interval that satisfies the safety factor by a three-dimensional analysis.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.163-167
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• 1988

The conditions for the protoplast fusion of auxotrophic mutants of Penicillium verruculosum were determined. A preparation of commercial enzyme Novozym 234 was used to successfully isolate protoplast from the 20hr old mycelium of P. verruculosum. Under optimal condition, the protoplast yield ranged from 2.4$\times$10$^7$ to 3.0$\times$10$^7$ protoplasts from 400mg of damp mycelia of various auxotrophic mutant strains. The regeneration frequency ranged from 26.6 to 42.4% and the spontaneous reversion frequency of the protoplasts on the regeneration minimal medium was less than 10$^7$. The optimal concentration of PEG 6000 was 20%, and exposure of protoplasts to PEG for 10 min was found to be sufficient for protoplast fusion. Optimal pH of fusion mixture was deter-mined as 5.5 and l0mM of calcium chloride in fusion mixture effectively enhanced the protoplast fusion frequency. Under optimal condition, the fusion frequency between various auxotrophs ranged from 1.8$\times$10$^{-3}$ to 3.5$\times$0$^{-3}$.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.156-162
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• 1988

Optimal conditions for the formation and regeneration of protoplasts of the cellulolytic fungus Penicillium verruculosum were investigated. Among the various commercial cell wall lytic enzymes tested, 0.5%(w/ v) Novozym 234 was the most effective for protoplast formation. The highest yield of protoplast exceeding 4.5$\times$10$^6$/m$\ell$ obtained when 400mg of 20 hr-old mycelia was incubated with 0.5%(w/v) Novozym 234 at 3$0^{\circ}C$ for 1 hr. The best osmotic stabilizer for the isolation and re-generation of protoplasts was 0.7M sorbital (pH 5.6) and 0.6M MgSO$_4$(pH 5.6), respectively. When 0.6M MgSO$_4$was added as osmotic stabilizer to the complete medium, the maximum regeneration frequency obtained was 4.6-27.8%. Micromorphological change of giant protoplasts into hyphae was observed during incubation in the regeneration liquid medium.

• Journal of Microbiology and Biotechnology
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• v.25 no.4
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• pp.496-502
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• 2015

Polyketide secondary metabolites share common precursor pools, acyl-CoA. Thus, the effects of engineering strategies for heterologous and native secondary metabolite production are often determined by the measurement of pikromycin in Streptomyces venezuelae. It is hard to compare the effectiveness of engineering targets among published data owing to the different pikromycin production media used from one study to the other. To determine the most important nutritional factor and establish optimal culture conditions, medium optimization of pikromycin from Streptomyces venezuelae ATCC 15439 was studied with a statistical method, Plackett-Burman design. Nine variables (glucose, sucrose, peptone, (NH₄)₂SO₄, K₂HPO₄, KH₂PO₄, NaCl, MgSO₄·7H₂O, and CaCO₃) were analyzed for their effects on a response, pikromycin. Glucose, K₂HPO₄, and CaCO₃ were determined to be the most significant factors. The path of the steepest ascent and response surface methodology about the three selected components were performed to study interactions among the three factors, and the fine-tune concentrations for maximized product yields. The significant variables and optimal concentrations were 139 g/1 sucrose, 5.29 g/l K₂HPO₄, and 0.081 g/l CaCO₃, with the maximal pikromycin yield of 35.5 mg/l. Increases of the antibiotics production by 1.45-fold, 1.3-fold, and 1.98-fold, compared with unoptimized medium and two other pikromycin production media SCM and SGGP, respectively, were achieved.

• Journal of Microbiology and Biotechnology
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• v.22 no.5
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• pp.628-636
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• 2012

Raw-starch-digesting enzyme (RSDA) was immobilized on Amberlite beads by conjugation of glutaraldehyde/polyglutaraldehyde (PG)-activated beads or by crosslinking. The effect of immobilization on enzyme stability and catalytic efficiency was evaluated. Immobilization conditions greatly influenced the immobilization efficiency. Optimum pH values shifted from pH 5 to 6 for spontaneous crosslinking and sequential crosslinking, to pH 6-8 for RSDA covalently attached on polyglutaraldehyde-activated Amberlite beads, and to pH 7 for RSDA on glutaraldehyde-activated Amberlite. RSDA on glutaraldehyde-activated Amberlite beads had no loss of activity after 2 h storage at pH 9; enzyme on PG-activated beads lost 9%, whereas soluble enzyme lost 65% of its initial activity. Soluble enzyme lost 50% initial activity after 3 h incubation at $60^{\circ}C$, whereas glutaraldehyde-activated derivative lost only 7.7% initial activity. RSDA derivatives retained over 90% activity after 10 batch reuse at $40^{\circ}C$. The apparent $K_m$ of the enzyme reduced from 0.35 mg/ml to 0.32 mg/ml for RSDA on glutaraldehyde-activated RSDA but increased to 0.42 mg/ml for the PG-activated RSDA derivative. Covalent immobilization on glutaraldehyde Amberlite beads was most stable and promises to address the instability and contamination issues that impede the industrial use of RSDAs. Moreover, the cheap, porous, and non-toxic nature of Amberlite, ease of immobilization, and high yield make it more interesting for the immobilization of this enzyme.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1605-1613
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• 2010

Twenty-nine P. polymyxa strains isolated from rhizospheres of various crops were clustered into five genotypic groups on the basis of BOX-PCR analysis. The characteristics of several plant growth-promoting factors among the isolates revealed the distinct attributes in each allocated group. Under gnotobiotic conditions, inoculation of pepper roots with P. polymyxa isolates significantly increased the biomass in 17 of total 29 treated plants with untreated plants. Experiments on induced systemic resistance (ISR) against bacterial spot pathogen Xanthomonas axonopodis pv. vesicatoria in pepper by P. polymyxa strains were conducted and only one isolate (KNUC265) was selected. Further studies into ISR mediation by the KNUC265 strain against the soft-rot pathogen Erwinia carotovora subsp. carotovora in tobacco demonstrated that the tobacco seedlings exposed to either bacterial volatiles or diffusible metabolites exhibited a reduction in disease severity. In conclusion, ISR and plant growth promotion triggered by P. polymyxa isolates were systemically investigated on pepper for the first time. The P. polymyxa KNUC265 strain, which elicited both ISR and plant growth promotion, could be potentially used in improving the yield of pepper and possibly of other crops.

• Journal of Microbiology and Biotechnology
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• v.23 no.3
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• pp.351-356
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• 2013

A high ${\beta}$-glucosidase (BGL)-producing strain, Stereum hirsutum, was identified and isolated and showed a maximum BGL activity (10.4 U/ml) when cultured with Avicel and tryptone as the carbon and nitrogen sources, respectively. In comparison with other BGLs, BGL obtained from S. hirsutum showed a higher level of activity to cellobiose ($V_{max}$ = 172 U/mg, and $k_{cat}$ = 281/s). Under the optimum conditions (600 rpm, $30^{\circ}C$, and pH 6.0), the maximum BGL activity of 10.4 U/ml with the overall productivity of 74.5 U/l/h was observed. BGL production was scaled up from a laboratory scale (7-L fermenter) to a pilot scale (70-L fermenter). When S. hirsutum was cultured in fed-batch culture with rice straw as the carbon source in a 70-L fermenter, a comparable productivity of 78.6 U/l/h was obtained. Furthermore, S. hirsutum showed high levels of activity of other lignocellulases (cellobiohydrolase, endoglucanase, xylanase, and laccase) that are involved in the saccharification of biomasses. Application of S. hirsutum lignocellulases in the hydrolysis of Pinus densiflora and Catalpa ovata showed saccharification yields of 49.7% and 43.0%, respectively, which were higher than the yield obtained using commercial enzymes.

• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.137-143
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• 2013

The diversity and distribution of methanogenic archaea in a four-compartment anaerobic baffled reactor (ABR) treating sugar refinery wastewater were investigated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). At an organic loading rate of 5.33 kg $COD/m^3{\cdot}day$, the ABR could perform steadily with the mean chemical oxygen demand (COD) removal of 94.8% and the specific $CH_4$ yield of 0.21 l/g $COD_{removed}$. The $CH_4$ content in the biogas was increased along the compartments, whereas the percentage of $H_2$ was decreased, indicating the distribution characteristics of the methanogens occurred longitudinally down the ABR. A high phylogenetic and ecological diversity of methanogens was found in the ABR, and all the detected methanogens were classified into six groups, including Methanomicrobiales, Methanosarcinales, Methanobacteriales, Crenarchaeota, Arc I, and Unidentified. Among the methanogenic population, the acid-tolerant hydrogenotrophic methanogens including Methanoregula and Methanosphaerula dominated the first two compartments. In the last two compartments, the dominant methanogenic population was Methanosaeta, which was the major acetate oxidizer under methanogenic conditions and could promote the formation of granular sludge. The distribution of the hydrogenotrophic (acid-tolerant) and acetotrophic methanogens in sequence along the compartments allowed the ABR to perform more efficiently and steadily.

• Journal of Korean Society of Environmental Engineers
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• v.36 no.10
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• pp.704-710
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• 2014

Biomass is considered an alternative energy which can solve an greenhouse gas problem like $CO_2$ which is a major contributor to global warming. The biomass can be converted to various energy sources through thermochemical conversion. In this study, a continuous gasifier was engineered for a wood biomass gasification. The biomass was used a waste wood. The experiments of $CO_2$ gasification were achieved as the gasification temperature, moisture content and input $CO_2$ concentration. The results showed that the yield of producer gas increased with an increasing the gasification temperature. The amount of the light tar increased due to the decomposition of gravimetric tar by the thermal cracking, and the char was confirmed pore development through the SEM analysis. The CO concentration was increased with an increased input $CO_2$ concentration from Boudouard reaction. Through the parametric screening studies, the hydrogen and carbon monoxide concentration were 32.91% and 48.33% at the optimal conditions of this test rig.