• Journal of Microbiology and Biotechnology
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• v.31 no.6
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• pp.823-832
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• 2021

Mushroom cultivation along with the palm oil industry in Malaysia have contributed to large volumes of accumulated lignocellulosic residues that cause serious environmental pollution when these agroresidues are burned. In this study, we illustrated the utilization of lignocellulolytic enzymes from the spent mushroom substrate of Pleurotus pulmonarius for the hydrolysis of palm oil mill effluent (POME). The hydrolysate was used for the production of biohydrogen gas and enzyme assays were carried out to determine the productivities/activities of lignin peroxidase, laccase, xylanase, endoglucanase and β-glucosidase in spent mushroom substrate. Further, the enzyme cocktails were concentrated for the hydrolysis of POME. Central composite design of response surface methodology was performed to examine the effects of enzyme loading, incubation time and pH on the reducing sugar yield. Productivities of the enzymes for xylanase, laccase, endoglucanase, lignin peroxidase and β-glucosidase were 2.3, 4.1, 14.6, 214.1, and 915.4 U g^-1, respectively. A maximum of 3.75 g/lof reducing sugar was obtained under optimized conditions of 15 h incubation time with 10% enzyme loading (v/v) at a pH of 4.8, which was consistent with the predicted reducing sugar concentration (3.76 g/l). The biohydrogen cumulative volume (302.78 ml H₂.L^-1 POME) and 83.52% biohydrogen gas were recorded using batch fermentation which indicated that the enzymes of spent mushroom substrate can be utilized for hydrolysis of POME.

• Journal of the Semiconductor & Display Technology
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• v.18 no.1
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• pp.32-37
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• 2019

Titanium dioxide ($TiO_2$) is a promising dielectric material in the semiconductor industry for its high dielectric constant. However, for utilization on Si substrate, $TiO_2$ film meets with a difficulty due to the large leakage currents caused by its small conduction band energy offset from Si substrate. In this study, we propose an in-situ plasma oxidation process in plasma-enhanced atomic layer deposition (PE-ALD) system to form an oxide barrier layer which can reduce the leakage currents from Si substrate to $TiO_2$ film. $TiO_2$ film depositions were followed by the plasma oxidation process using tetrakis(dimethylamino)titanium (TDMAT) as a Ti precursor. In our result, $SiO_2$ layer was successfully introduced by the plasma oxidation process and was used as a barrier layer between the Si substrate and $TiO_2$ film. Metal-oxide-semiconductor ($TiN/TiO_2/P-type$ Si substrate) capacitor with plasma oxidation barrier layer showed improved C-V and I-V characteristics compared to that without the plasma oxidation barrier layer.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.419-428
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• 2021

To efficiently recycle GH78 thermostable rhamnosidase (TpeRha) and easily separate it from the reaction mixture and furtherly improve the enzyme properties, the magnetic particle Fe3O4-SiO2-NH2-Cellu-ZIF8 (FSNcZ8) was prepared by modifying Fe3O4-NH2 with tetraethyl silicate (TEOS), microcrystalline cellulose and zinc nitrate hexahydrate. FSNcZ8 displayed better magnetic stability and higher-temperature stability than unmodified Fe3O4-NH2 (FN), and it was used to adsorb and immobilize TpeRha from Thermotoga petrophilea 13995. As for properties, FSNcZ8-TpeRha showed optimal reaction temperature and pH of 90℃ and 5.0, while its highest activity approached 714 U/g. In addition, FSNcZ8-TpeRha had better higher-temperature stability than FN. After incubation at 80℃ for 3 h, the residual enzyme activities of FSNcZ8-TpeRha, FN-TpeRha and free enzyme were 93.5%, 63.32%, and 62.77%, respectively. The organic solvent tolerance and the monosaccharides tolerance of FSNcZ8-TpeRha, compared with free TpeRha, were greatly improved. Using naringin (1 mmol/l) as the substrate, the optimal conversion conditions were as follows: FSNcZ8-TpeRha concentration was 6 U/ml; induction temperature was 80℃; the pH was 5.5; induction time was 30 min, and the yield of products was the same as free enzyme. After repeating the reaction 10 times, the conversion of naringin remained above 80%, showing great improvement of the catalytic efficiency and repeated utilization of the immobilized α-L-rhamnosidase.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.4
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• pp.275-287
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• 2000

Fermentation strategies for recombinant protein production in Pichia pastoris have been investigated and are reviewed here. Characteristics of the expression system, such as phenotypes and carbon utilization, are summarized. Recently reported results such as growth model establishment, app58lication of a methanol sensor, optimization of substrate feeding strategy, DOstat controller design, mixed feed technology, and perfusion and continuous culture are discussed in detail.

• Journal of the Korean Wood Science and Technology
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• v.17 no.3
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• pp.52-60
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• 1989

This study was carried out to know the possibility of simultaneous utilization of laccase from white-rot fungus with cellulase on enzymatic hydrolysis of cellulosic substrate from autohydrolyzed oak wood. Laccases from 3 white-rot fungi, Pleurotus ostreatus. Ganoderma lucidum, and Phanerochaete chrysosporium, were isolated, purified and measured their activities. The highest activity was shown in Pleurotus ostreatus and the lowest in Phanerochaete chrysosporium. Laccase from Pleurotus ostreatus has optimum pH of 5.94, Km value of 3.209 mM and appeared to be stable at relatively wide pH range, 4.7-8.72. Temperature stability showed that 60% activity was preserved after 40 minutes at $50^{\circ}C$. Laccase from Ganoderma lucidum reached to the maximum activity during 15-20 day incubation. This enzyme has optimum pH of 6.45, Km value of 6.71 mM and pH range of 5.0-9.0 for stabilization. 95% activity was preserved at $30^{\circ}C$ and 58% activity at $50^{\circ}C$. Concerned to the enzymatic hydrolysis of cellulosic substrate with both enzymes, cellulase and laccase, simultaneously, mixed culture filtrates and mycellium extracts were shown higher hydrolysis rates than those of Trichoderma viride. There were no significant differences in the extent of hydrolysis among various mixed culture filtrates and mycellium extracts.

• Microbiology and Biotechnology Letters
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• v.4 no.4
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• pp.152-158
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• 1976

Cultivation condition of yeast on the utilization of fermentable substrate from the cellulosic wastes such as rice hull, rice straw and corn starch cake was investigated. The results obtained were summarized as follows；1. Corn starch cake was respectively added to rice hull and rice straw in order to increase sugar concentration in the hydrolyzate, and then hydrolyzed. As the result, concentration of sugar in hydrolyzed solution of rice hull was 9.12％, in that of rice straw was 7.98％. 2. It was found that calcium carbonate as a neutralizer was the most effective to prepare the culture broth of yeast. 3. An optimal growth of Hansenula subpelliculosa GFY-2 was observed in the medium prepared by adding 0.3％ of ammonium sulfate, 0.4％ of potassium phosphate dibasic, 0.02％ of magnesium sulfate, sodium chloride and calcium chloride to hydrolyaed sugar solution, respectively. 4. Hansenula subpellicuiosa GFY-2 cultured in the substrate solution which of rice hull and rice straw added to corn starch cake was assimilated more than 90％ of sugar in the hydrolyzate within 48 hours. The yeast cells yielded in rice hull was 46.5％, and that of rice straw 45.4％ to utilized sugars.

• BMB Reports
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• v.45 no.2
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• pp.59-70
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• 2012

Carbon catabolite repression (CCR) is a key regulatory system found in most microorganisms that ensures preferential utilization of energy-efficient carbon sources. CCR helps microorganisms obtain a proper balance between their metabolic capacity and the maximum sugar uptake capability. It also constrains the deregulated utilization of a preferred cognate substrate, enabling microorganisms to survive and dominate in natural environments. On the other side of the same coin lies the tenacious bottleneck in microbial production of bioproducts that employs a combination of carbon sources in varied proportion, such as lignocellulose-derived sugar mixtures. Preferential sugar uptake combined with the transcriptional and/or enzymatic exclusion of less preferred sugars turns out one of the major barriers in increasing the yield and productivity of fermentation process. Accumulation of the unused substrate also complicates the downstream processes used to extract the desired product. To overcome this difficulty and to develop tailor-made strains for specific metabolic engineering goals, quantitative and systemic understanding of the molecular interaction map behind CCR is a prerequisite. Here we comparatively review the universal and strain-specific features of CCR circuitry and discuss the recent efforts in developing synthetic cell factories devoid of CCR particularly for lignocellulose-based biorefinery.

• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 2003.04a
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• pp.188-191
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• 2003

Sing]e-well-push-pull tests were developed for use in assessing the feasibility of in-situ aerobic cometabolism of chlorinated aliphatic hydrocarbons (CAHs). The series includes Transport tests, Biostimulation tests, and Activity tests. Transport tests are conducted to evaluate the mobility of solutes used in subsequent tests. These included bromide or chloride (conservative tracers), propane (growth substrate), ethylene, propylene (CAH surrogates), dissolved oxygen (electron acceptor) and nitrate (a minor nutrient). Tests were conducted at an experimental well field of Oregon State University. At this site, extraction phase breakthrough curves for all solutes were similar, indicating apparent conservative transport of the dissolved gases and nitrate prior to biostimulation. Biostimulation tests were conducted to stimulate propane-utilizing activity of indigenous microorganisms and consisted of sequential injections of site groundwater containing dissolved propane and oxygen. Biostimulation was detected by the increase in rates of propane and oxygen utilization after each injection. Activity tests were conducted to quantify rates of substrate utilization and to confirm that CAH-transforming activity had been stimulated. In particular, the transformation of injected CAH surrogates ethylene and propylene to the cometabolic byproducts ethylene oxide and propylene oxide provided evidence that activity of the monooxygenase enzyme system, responsible for aerobic cometabolic transformations of CAHs had been stimulated. Estimated zero-order transformation rates decreased in the order propane > ethylene > propylene. The series of push-pu3l tests developed and field tested in this study should prove useful for conducting rapid, low-cost feasibility assessments for in situ aerobic cometabolism of CAHs.

• Korean Journal of Food Science and Technology
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• v.52 no.2
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• pp.144-148
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• 2020

Corn silk (Okmi-su) was anciently adopted as a material for tea or beverage. Corn silk extracts (CSE) contain bioactive phytochemicals such as phenolic acid, flavonoids, ascorbic acid, tannins, and glycosides. Under the impact of these functional components, CSE has benefits for antioxidation, diuresis, anti-diabetes, and dyslipidemia recovery. Nonetheless, its role in whole-body adiposity was not investigated; therefore, the effects of CSE on obesity were evaluated in high-fat diet-induced obese mice. Mice were assigned to either group (n=12); 1) normal diet (18% kcal from fat), 2) high-fat diet (45% kcal from fat, the control), 3) high-fat diet with CSE (800 mg/kg diet), and 4) high-fat diet with orlistat (500 mg/kg diet, a comparable control for weight loss). Our results showed that body weight, adiposity, and energy expenditure in obese mice were not altered by CSE. Lean body mass tended to decrease by CSE, which can be explained by stimulation of diuresis (p=0.06). In conclusion, our results suggest that dietary consumption of CSE does not influence the adiposity and underlying substrate utilization in high-fat diet-induced obese mice.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.577-586
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• 2019

The engineered Aspergillus oryzae has a high NADPH demand for xylose utilization and overproduction of target metabolites. Glucose-6-phosphate dehydrogenase (G6PDH, E.C. 1.1.1.49) is one of two key enzymes in the oxidative part of the pentose phosphate pathway, and is also the main enzyme involved in NADPH regeneration. The open reading frame and cDNA of the putative A. oryzae G6PDH (AoG6PDH) were obtained, followed by heterogeneous expression in Escherichia coli and purification as a his6-tagged protein. The purified protein was characterized to be in possession of G6PDH activity with a molecular mass of 118.0 kDa. The enzyme displayed maximal activity at pH 7.5 and the optimal temperature was $50^{\circ}C$. This enzyme also had a half-life of 33.3 min at $40^{\circ}C$. Kinetics assay showed that AoG6PDH was strictly dependent on $NADP^+$ ($K_m=6.3{\mu}M$, $k_{cat}=1000.0s^{-1}$, $k_{cat}/K_m=158.7s^{-1}{\cdot}{\mu}M^{-1}$) as cofactor. The $K_m$ and $k_{cat}/K_m$ values of glucose-6-phosphate were $109.7s^{-1}{\cdot}{\mu}M^{-1}$ and $9.1s^{-1}{\cdot}{\mu}M^{-1}$ respectively. Initial velocity and product inhibition analyses indicated the catalytic reaction followed a two-substrate, steady-state, ordered BiBi mechanism, where $NADP^+$ was the first substrate bound to the enzyme and NADPH was the second product released from the catalytic complex. The established kinetic model could be applied in further regulation of the pentose phosphate pathway and NADPH regeneration of A. oryzae to improve its xylose utilization and yields of valued metabolites.