• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1267-1272
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• 2005

The effects of coexpression of GroEL/ES and DnaK/DnaJ/GrpE chaperones on the productivity of the soluble form of human granulocyte colony stimulating factor (hG-CSF) in E. coli were examined. Recombinant hG-CSF protein was coexpressed with DnaK/DnaJ/GrpE or GroEL/ES chaperones under the control of the araB or Pzt-1 promoter, respectively. The optimal concentration of L-arabinose for the expression of DnaK/DnaJ/GrpE was found to be 1 mg/ml. When L-arabinose was added at $OD_{600}$=0.2 (early-exponential phase), soluble hG-CSF production was greatly increased. In addition, it was observed that the DnaK/DnaJ/GrpE and GroEL/ES chaperones had no synergistic effects on preventing aggregation of hG-CSF protein. Consequently, by coexpression of the DnaK/DnaJ/GrpE chaperone, the signal intensity of the hG-CSF protein band in the soluble fraction of cell lysate was increased from $3.5\%\;to\;13.9\%$, and Western blot analysis also revealed about a 4-5-fold increase of production of soluble hG-CSF over the non-induction case of DnaK/DnaJ/GrpE.

• Journal of Korean Society on Water Environment
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• v.23 no.4
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• pp.498-503
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• 2007

The performance of elutriated acid fermentation to evaluate the effects of microwave irradiation and pH control as pretreatment was investigated. The MW pH 7 reactor which was used the pretreated primary sludge as microwave irradiation was operated at pH 7 and $35^{\circ}C$. The EAF pH 9 reactor was operated at pH 9 and $35^{\circ}C$ without pretreatment. The SCOD and VFAs production rate were 0.17 gSeOD/gVSrem. and 0.27 gVFAs as COD/gVSrem. in MW pH 7 reactor, 0.16 gSCOD/gVSrem. and 0.24 gVFAs as COD/gVSrem. in EAF pH 9 reactor, respectively. VS and Volume reduction were 54% and 48% in MW pH 7 reactor, 54.6% and 36% in EAF pH 9 reactor, respectively. A comparison of the microwave irradiation and controlled pH in elutriated acid fermentation showed that the former is more efficient in SCOD and VFAs production and it rises to slightly higher reduction in the volume of the sludge. In addition, E. coli. was not detected in the wasting sludge of MW pH 7 reactor. Based on the results, microwave irradiation appeared to be one of the viable options for generating class A sludge. According to the batch tests, sequencing batch test which was used the pretreated primary sludge as microwave was performed at pH 7 and $35^{\circ}C$, SCOD production was 0.16 gSCOD/gVSrem., VS reduction and volume reduction were 64% and 63%, respectively.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.1
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• pp.68-78
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• 1997

Laboratory and greenhouse studies were conducted to determine differential sensitivities on absorption of $^{14}$ C-oxyfluorfen and the anatomical responses in wheat and barley to protoporphyrinogen oxidase-inhibiting herbicides [oxyfluorfen (2-chloro-1- (3-ethoxy -nitrophen-oxy)-4-(trifluoromethyl) benzene, acifluorfen(5-[2-chloro-4-(trifluoromethyl) phenoxy]-2-nitro-benzoic acid), bifenox(methyl-5-(2, 4-dichlorophenoxy)-2-nitrobenzoate) and oxadiazon(5-tert-butyl-3-(2, 4-dichloro-5-isopropoxyphenyl)-1, 3, 4-oxadiazol-2-one)]. I$_{50}$ value of the tolerant wheat cultivars to oxyfluorfen was about 10$^{-4}$ , whereas that of the susceptible barley cultivars was about 10$^{-6}$ M, showing significant difference between the two groups. When foliage were applied with acifluorfen, bifenox or oxadiazon, the oxyfluorfen-tolerant wheat showed less decreased in shoot fresh weight and chlorophyll content than the susceptible barley. Also, when soil-applied with these herbicides test plants showed similar tendency in foliar application. Electrolyte leakage from the tissue treated with these compounds was the more influenced in the barley than the wheat. Malondialdehyde(MDA) production as index of lipid peroxidation was greater in the barley than the wheat by treatment of these compounds. Therefore, the differential sensitivities of wheat and barley to protoporphyrinogen oxidaseinhibiting herbicides was showed by our greenhouse and in vitro experiment. The absorption rates of $^{14}$ C-oxyfluorfen were higher in the barley than the wheat. And this tendency was showed appararitly difference by increase of treatment durations. After the oxfluorfen and oxadiazon treatment, the tolerant wheat did not show the structural damage in leaf surface, but the susceptible barley was damaged in the leaf waxy layer. However, the acifluorfen and bifenox treatment showed no difference between wheat and barley. The anatomical changes by these compounds treatment were not observed in the tolerant wheat but epidermal cell and mesophyll cell were highly broken in the susceptible barley.

• Journal of Life Science
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• v.22 no.4
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• pp.508-515
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• 2012

In this study, DEAE-cotton [derivatized by 2-(diethylamino)ethyl chloride hydrochloride (DEAE HCl)] was prepared as a carrier for immobilized $Pichia$ $stipitis$ for ethanol production. When cotton was derivatized with 0.5 M DEAE HCl, the yeast cell suspension was adsorbed at 100% of the initial cell $OD_{600}$. The adsorbed yeast cells were estimated to be 101.8 mg-dry cells/g-DEAE-cotton. In particular, when a flask culture using the immobilized yeast cells was conducted in a glucose and xylose-containing medium, the yeast cells on the DEAE-cotton gradually produced ethanol, according to glucose and xylose consumption; the ethanol yield was approximately 0.33 g-ethanol/g-monosaccharide. Because DEAE-cotton was successfully used as a carrier for ethanol production from a glucose and xylose-containing medium, we expect that this bioethanol production process may be used for the bioethanol production process from the hydrolysate of lignocellulosic biomass. All the results of DEAE-cotton were compared with those of DEAE-cellulose as a carrier for immobilization.

• Journal of Microbiology and Biotechnology
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• v.21 no.12
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• pp.1264-1269
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• 2011

Interleukin-2 (IL-2) is a vital cytokine secreted by activated T lymphocytes, and plays an important role in the regulation of cellular functions and immunity of animals. In this study, the recombinant duck IL-2 (rduIL-2) was secretory expressed in Pichia pastoris (P. pastoris). The recombinant P. pastoris strain was cultured in shake flasks and then scaled up in a 5.0-l bioreactor. The result showed that the maximal fresh-cell-weight of 594.1 g/l and the maximal $OD_{600}$ of 408 were achieved in the bioreactor. The rduIL-2 was purified by two steps of purification procedures, and approximately 311 mg of rduIL-2/L fermentation supernatant was obtained. SDS-PAGE showed that the purified rduIL-2 constituted a homogeneous band of ~16 kDa or ~14 kDa corresponding to the glycosylated or non-glycosylated duIL-2 protein in size, respectively. The bioactivity of rduIL-2 was determined by lymphocyte proliferation assay. The result indicated that the rduIL-2 greatly promoted the proliferation of ConA-stimulated lymphocytes in vitro. The P. pastoris expression system described here could provide promising, inexpensive, and large-scale production of the rduIL-2, which lays the foundation for development of novel immunoadjuvants to enhance both the immunity of ducks against various infectious pathogens and vaccine efficacy.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.344-349
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• 2004

A gene encoding inulin-degrading enzyme of Bacillus sp. snu-7 with ORF of 1536 nucleotides was cloned. And it was overexpressed as His-tagged protein in E. coli BL21(DE3) pLysS using pRSET B vector containing mature enzyme sequence. Maximum enzyme production was achieved by IPTG (0.1 mM) induction at $OD_{600}$ 1.2 and $30^{\circ}C$ followed by 6 h incubation. The expressed protein purified through immobilized metal affinity chromatography showed molecular mass of 60 kDa on SDS-PAGE. Results of thin-layer chromatography using inulin as a substrate showed the enzyme to be an exotype inulinase capable of producing only monomeric fructose as a product. $K_m$ and $k_{cat}$, for the hydrolyses of inulin and sucrose were $2.28\pm0.08$ mM and 358.05$\pm$20.38 $min^{-l}$, and 22.02$\pm$0.41 mM and 4619.11$\pm$215.12 $$min^{-1}, respectively. Optimal activity of the exoinulinase occurred at pH 7.0 and $50^{\circ}C$.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 1994.10a
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• pp.449-454
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• 1994

The Stewart Platform is one example of a motion simulator which generater 6DOF motion in space by six actuators in parallel. The presented control methrol of 6DOF motion simulator is generally classified into two types, one is SISO and the other is MIMO control type. The SISO control can't compensate for external load variation and different dynamic behavior of 6DOF motion, trerefore this type don's control motion precisely. On the other hand, the MIMO control compensates for a interference of 6DOF motion because MIMO controller is designed with 6DOF motion simulator synamics. But MIMO control of motion simulator has a complexity of 6DOF displacement feedback, because in oder to obtain feedback value we must solve the forward kinematics using measurement of cylinder length or design a state estimator, unless measurement of 6DOF displacement is possible. In this paper, a multivariable controller using H .inf. optimal control theory is designed to consider a interference of 6DOF motion and to obtain robust,precise control of system. Also in order to solve the mentioned problem of MIMO control, this paper presents a modified MIMO control model which control 6DOF motion by using feedback of measurement od cylinder length.

• BMB Reports
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• v.38 no.3
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• pp.294-299
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• 2005

In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were successfully expressed upon methanol induction. The absorbance ($OD_{600}$) of the broth can reach 350~500 at the end of fed-batch phase. After the induction, the expression level of the recombinant Fab (soluble) reached 420~458 mg/L. The recombinant Fab fragment was purified from the crude culture supernatant by ion exchange chromatography and the purity of the recombinant Fab fragment was over 95%. The affinity activities of the crude fermentation supernatant and the purified Fab were analyzed by indirect ELISA, which showed that the purified recombinant Fab fragment had high affinity activity with hepatitis B surface antigen.

• Journal of Apiculture
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• v.33 no.4
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• pp.277-282
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• 2018

Ethanol extracted propolis (EEP) was mixed with honey (honeypolis) to dissolve well in water and in vitro skin irritation test was conducted. In vitro method is designed to predict and classify the skin irritation potential of a chemical by assessment of its effect on $EpiDerm^{TM}$, a reconstituted three-dimensional human epidermis model. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT after a 60 min exposure period. In this study under the given conditions honeypolis showed no irritant effects. Honeypolis meets acceptance criteria if: mean absolute OD 570 nm of the three negative control tissues is ${\geq}0.8$ and ${\leq}2.8$, mean relative tissue viability of the three positive control tissues is ${\leq}20%$, standard deviation of relative tissue viability obtained from each three concurrently tested tissues is ${\leq}18%$. Honeypolis is therefore classified as "non-irritant" in accordance with UN GHS "No Category".

• Journal of fish pathology
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• v.34 no.1
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• pp.111-115
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• 2021

Immunoglobulin M (IgM) of sevenband grouper (Epinephelus septemfasciatus) was purified by mannan-binding protein (MBP) affinity column. The purified IgM had an apparent molecular weights of 76 (heavy chain) and 28 (light chain) kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Eight hybridoma clones secreting monoclonal antibodies (mAbs) against sevenband grouper IgM were established. Antibody detection enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the 8 mAbs revealed that optical density (OD) values were clearly different between sera from BSA-immunization and non-immunization of sevenband grouper. These results suggest that the produced mAbs in this study are specifically reacted with IgM of sevenband grouper.