• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.344-357
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• 2020

Strain improvement and bioprocess development were undertaken to enhance hyaluronic acid(HA) production by Streptococcus zooepidemicus cells. Using a high-yielding mutant strain, statistical medium optimization was carried out in shake flask cultures, resulting in 52% increase in HA production (5.38 g/l) at the optimal medium composition relative to the parallel control cultures. For sufficient supply of dissolved oxygen (DO), which turned out to be crucial for enhanced production of HA, agitation system and speed were intensively investigated in 5 L bioreactor cultures. Increase in oxygen mass transfer coefficient (k_La) through increment of agitation speed (rpm) and 35% expansion of diameter of the newly-designed impellers showed significantly positive effects on HA production. By installing an expanded Rushton-turbine impeller for efficient break-down of sparged air, and an extended marine impeller above the Rushton-turbine impeller for efficient mixing of the air-born viscous fermentation broth, maximum amount of HA (9.79 g/l) was obtained at 450 rpm, 1.8 times higher level than that of the corresponding flask culture. Subsequently, the possibility of bioprocess scale-up to a 50 L bioreactor was investigated. Despite almost identical maximum HA production (9.11 vs 9.25 g/l), the average HA volumetric productivity (r_p) of the 50 L culture turned out only 74% compared to the corresponding 5 L culture during the exponential phase, possibly caused by shear damages imposed on the producing cells at the high stirring in the 50 L culture. The scale-up process could be successfully achieved if a scale-up criterion of constant oxygen mass transfer coefficient (k_La) is applied to the 50 L pilot-scale bioreactor system.

• Food Science and Preservation
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• v.10 no.1
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• pp.16-22
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• 2003

This study was conducted to find out effective MA (Modified Atmosphere) gas compositions on the oyster mushroom through statistical analysis of the respiration rate and MA storage for the various packaging materials. Under the various gas compositions, the oxygen consumption rate of oyster mushroom was from 28.9 to 161.4mgO$_2$/kg$.$hr and the carbon dioxide evolution rate was from 53.4 to 166.9 mgCO$_2$/kg$.$hr at 20$^{\circ}C$. The estimated MA condition of oyster mushroom were 2.5∼4.5%O$_2$and 11.5∼l3%CO$_2$by the RSREG(Response Surface Regression). The gas compositions of MA packaging are following that 0.03mm LDPE were 1.6∼3.0%O$_2$and 3.9∼5.3%CO$_2$,0.05mm LDPE were 1.2∼1.3%O$_2$and 9.0∼11.1%CO$_2$and Nylon+PE were 0.9∼1.2%O$_2$and 33.5∼39.6%CO$_2$. The weight loss increased at 0.03mm LDPE but has the lowest value at Nylon+PE. The hardness of pileus and stipe was decreased with storage periods. The $\Delta$E-value increased with storage period and seriously changed in early storage period at 12 and 20$^{\circ}C$. In the 0.05mm LDPE, the gas compositions of packaging were similar to estimated gas compositions from the RSREG and the storage quality was superior to the other packaging materials in weight loss, hardness, and color difference at 4, 12 and 20 $^{\circ}C$.

• Food Science and Preservation
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• v.17 no.2
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• pp.281-289
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• 2010

We determined the optimum ethanolic conditions for extraction of $\gamma$-oryzanol and other functional components from rice bran, using response surface methodology (RSM). A central composite design was used to investigate the effects of the independent variables of solvent ratio ($X_1$), extraction temperature ($X_2$), and extraction time ($X_3$), on dependent variables including yield ($Y_1$), total phenolic content ($Y_2$), electron-donating activity ($Y_3$), ferulic acid level ($Y_4$), and $\gamma$-oryzanol concentration ($Y_5$). Solvent ratio and extraction temperature were the most important factors in extraction. The maximum yield was at 22.56 mL/g ($X_1$), 78.19C ($X_2$), and 522.15 min ($X_3$), at the saddle point. Total phenolic levels were little affected by solvent ratio or extraction temperature. The maximum concentration of extracted total phenolics was 90.78mg GAE/100 g at 21.26 mL/g, $94.65^{\circ}C$, and 567.97 min. A maximum electron-donating ability of 54.72% was obtained with the parameters 20.20 mL/g,$81.89^{\circ}C$, and 701.87 min, at the highest point. The maximum level of ferulic acid components was 210.47 mg/100g at 5.22 mL/g, $79.66^{\circ}C$, and 575.24 min. In addition, the maximum $\gamma$-oryzanol concentration was 660.39 mg/100g at 5.10 mL/g, $81.83^{\circ}C$, and 587.39 min. The optimum extraction conditions were a solvent ratio of 10.45 mL/g, $80^{\circ}C$ extraction temperature, and 535 min extraction time. Predicted extraction levels under optimized conditions were in line with experimental values.

• Journal of the Mineralogical Society of Korea
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• v.27 no.4
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• pp.301-310
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• 2014

To investigate the role of fault gauge in the behavior of heavy metals caused by the acid rock drainage in the area of pyrite-rich andesite, XRD, pH measurement, XRF, SEM-EDS, ICP, and sequential extraction method were used. Bed rock consists of quartz, pyrophyllite, pyrite, illite, and topaz, but the brown-colored fault gouge is composed of quartz, illite, chlorite, smectite, goethite, and cacoxenite. The mineral composition of bed rock suggests that it is heavily altered by hydrothermal activity. The concentrations of heavy metals in the bed rock are as follows, Zn > As > Cu > Pb > Cr > Ni > Cd, and those in fault gouge are As > Zn > Pb > Cr > Cu > Ni > Cd. The concentrations of the heavy metals in the fault gouge are generally higher than those in the bed rock, especially for Pb, As, and Cr, which were more than twice as those in the bed rock. It is believed that the difference in the amount of heavy metals between the bed rock and the fault gouge is mainly due to the existence of goethite which is the main mineral composition in the fault gouge and can play important role in sequestering these metals by coprecipitation and adsorption. The low pH, caused by oxidation of pyrite, also plays significant role in fixation of those metals. It is confirmed that the fractions of labile (step 1) and acid-soluble (step 2), which can be easily released into the environment, were higher in the bed rock. Those fractions were relatively low in fault gauge, suggesting that fault gauge can play important role as a sink of heavy metals to prevent those ones from being released in the area where the acid rock drainage can have an influence.

• Journal of the Mineralogical Society of Korea
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• v.22 no.1
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• pp.23-34
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• 2009

The adsorption of iodide on untreated bentonite and bentonites modified with organic cation (i.e., hexadecylpyridinium chloride monohydrate ($HDP^+$)) was investigated, and the organobentonites were characterized using uptake measurements, ${\mu}$-XRD, and electrophoretic mobilities measurement. Uptake measurements indicate that bentonite has a high affinity for $HDP^+$. Our ${\mu}$-XRD study indicates that organobentonites significantly expanded in basal spacing and organic cations were substantially intercalated into the interlayer spaces of bentonite. The electrophoretic mobility indicates that organobentonite tht is modified with organic cations in excess of the CEC of bentonite is completely different from untreated bentonite in the surface charge distribution. We found significant differences in adsorption capacities of iodide depending on the bentonite properties as follows: iodide adsorption capacities were 439 mmol/kg for the bentonite modified with $HDP^+$ at an equivalent amount corresponding to 200% of the CEC of bentonite whereas no adsorption of iodide was observed for the untreated bentonite. The molecular environments of iodine adsorbed on organobentonites were further studied using I K-edge and $L_{III}$-edge x-ray absorption spectroscopy (XAS). The X-ray absorption near-edge structure (XANES) of iodine spectra from organobentonites was similar to that of KI reference solution. Linear combination fitting of EXAFS data suggests the fraction of iodine reacted with the organic compound increased with increasing loading of the organic compound on organobentonites. In this study, we observed significant differences in the adsorption environments of iodide depending on the modified property of bentonite and suggest that an organobentonite has potential as reactive barrier material around a nuclear waste repository containing anionic radioactive iodide.

• Korean Journal of Materials Research
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• v.10 no.1
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• pp.97-106
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• 2000

In these respects the purpose of this research is manufacturing Co-W-P alloy thin film on the corning glass 2948 by electroless plating method using $NaH_2PO_2H_2O$ (sodium hypophosphite) as a reductant, and analyzing deposition rate, alloy composition, microstructure, and magnetic characteristics at various pH's and temperatures. For Co-P alloy thin film, the reductive deposition reaction occurred only in basic condition, not in acidic condition. The deposition rate increased as the pH and temperature increased, and the optimum condition was found at the pH of 10 and the temperature of 8$0^{\circ}C$. Also magnetic characteristics was found to be most excellent at the pH of 9 and the temperature of 7$0^{\circ}C$, resulting in the coercive force of 870Oe and the squareness of 0.78. At this condition, the contents of P was 2.54% and the thickness of the film was 0.216$\mu\textrm{m}$. For crystal orientation, we could not observe fcc for $\beta$-Co. On the other hand, (1010), (0002), (1011) orientation of hcp for $\alpha$-Co was observed. We could confirm the formation of longitudinal magnetization from dominant (1010) and (1011) orientation of Co-P alloy. For Co-W-P alloy thin film, coercive force was 500Oe and squareness was 0.6. For crystal orientation, (0002) orientation of $\alpha$-Co was dominatly found. Then we could confirm the formation of perpendicular magnetization. The content of P was constant at 0.8$\pm$0.2% and the content of W increased as the concentration of Na$_2$WO$_4$increased. When the concentration of Na$_2$WO$_4$was 0.1mol/L, the composition of W was 20%. We observed the changes of magnetic characteristics and microstructure of thin film depositions of Co-W-P by the heat treatment. For heat treatment, the temperature was increased step by step to 10$0^{\circ}C$, 20$0^{\circ}C$, 30$0^{\circ}C$, and 40$0^{\circ}C$ and it took 1 hour at each step in the reductive condition of hydrogen gas. By the heat treatment, flatness of surface was improved, but there were no changes on the magnetic characteristics and the microstructures.

• KSBB Journal
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• v.25 no.2
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• pp.142-154
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• 2010

The protein-bound innerpolysaccharides (IPS) produced by suspended mycelial cultures of Inonotus obliquus have promising potentials as an effective antidiabetic as well as an immunostimulating agents. To enhance IPS production, intensive strain improvement process should be carried out using large amount of UV-mutated protoplasts. During the whole strain-screening process, the stage of solid growth-culture was found to be the most time-requiring step, thus preventing rapid screening of high-yielding producers. In order to reduce the cell growth period in the solid growth-stage, therefore, solid growth-medium was optimized using the statistical methods such as (i) Plackett-Burman and fractional factorial designs (FFD) for selecting positive medium components, and (ii) steepest ascent (SAM) and response surface (RSM) methods for determining optimum concentrations of the selected components. By adopting the medium composition recommended by the SAM experiment, significantly higher growth rate was obtained in the solid growth-cultures, as represented by about 41% larger diameter of the cell growth circle and higher mycelial density. Sequential optimization process performed using the RSM experiments finally recommended the medium composition as follows: glucose 25.61g/L, brown rice 12.53 g/L, soytone peptone 12.53 g/L, $MgSO_4$ 5.53 g/L, and agar 20 g/L. It should be noted that this composition was almost similar to the medium combinations determined by the SAM experiment, demonstrating that the SAM was very helpful in finding out the final optimum concentrations. Through the use of this optimized medium, the period for the solid growth-culture could be successfully reduced to about 8 days from the previous 15~20 days, thus enabling large and mass screening of high producers in a relatively short period.

• KSBB Journal
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• v.22 no.5
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• pp.305-312
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• 2007

It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.

• KSBB Journal
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• v.22 no.5
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• pp.297-304
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• 2007

For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

• The Journal of Korean Academy of Prosthodontics
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• v.48 no.3
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• pp.209-214
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• 2010

Purpose: The purpose of this study was to investigate whether the re-osseointegration of the implants that had mechanical unscrewing possibly occurred or not. Furthermore, if it happened, the degree of re-osseointegration was evaluated by comparing with previous osseointegration. Materials and methods: The smooth implant (commercial pure titanium 99%) specimens, whose diameter and length was 3.75 mm, 4 mm, respectively were produced. Two implants were inserted into each tibia of 7 New Zealand female white rabbits weighing at least 3.0 kg. The torque removal force for each implant after 6 weeks of implants placement was measured and included in group I. The torque removal forces were assessed after the fixtures were re-screwed to original position and the subjects were allowed to have 4 more weeks for healing and included in group II. One rabbit was sacrificed after first measurement and produced 4 slide specimens in group I, and two rabbits were sacrificed after 2nd measurement, 7 slide specimens, in group II for histomorphologic investigations. All slide specimens were assessed based on the proportion of BIC (bone-implant contact) as well as CBa (Bone area in the cortical passage) value produced by counting the screw threads embedded in the compact bones under the optical microscopic analysis (${\times}20$). Statistical analysis was conducted to evaluate the torque removal force, BIC and CBa between group I and II. Results: As for the torque removal force, the result was $10.8{\pm}3.6$ Ncm for group I and $20.2{\pm}9.7$ Ncm for group II. Furthermore, the torque removal force of group II increased by 98.1% in average compared to group I (P<.05). On the other hand, histomorphologic analysis displayed that there was no statistical significance in BIC and CBa values between group I and the group II (P>.05), and RT/BIC and RT/CBa between group I and group II were statistically significant (P<.05). Conclusion: It is possible to obtain more substantial re-osseointegration within shorter periods than the period needed for the initial osseointegration in case of iatrogenically unscrewed implants.