• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1232-1238
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• 2004

To investigate the production and characteristics of thermostable levan sucrase from Rahnella aquatilis ATCC 33071, the levan sucrase gene from R. aquatilis was cloned and expressed in Escherichia coli without induction system. Expression of levansucrase gene in E. coli had no notable or detrimental effect on the growth of host strain, and the recombinant levan sucrase exhibited levan synthesis activity. Levansucrase was secreted to the periplasm in E. coli, and addition of $0.5\%$ glycine yielded further secretion of levansucrase to the growth medium and resulted in an increase of total levansucrase activity. Furthermore, the cellular levansucrase was evaluated for the production of levan by using toluene­permeabilized whole-cells. The levansucrase was thermostable at $37^{\circ}C$. The molecular size oflevan was $1{\times}\;10^{6}$ Da, as determined by HPLC, and the degree of polymerization of levan varied with incubation temperatures: Low incubation temperature was preferable for the production of high-molecular size levan. The present study demonstrated that the mass production of levan and levan oligosaccharides can be achieved by glycine supplementation to the growth medium or by toluene­permeabilized whole-cells.

• 한국수산과학회지
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• 제53권4호
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• pp.506-514
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• 2020

We investigated seed management and feeding regime of larval red spotted grouper Epinephelus akaara using live food organisms. Fertilized eggs were produced in March 2020 by manipulating photoperiod and water temperature. Broodstock (F0) produced approximately 0.30×10⁶ eggs, of which 0.25×10⁶ were fertilized; first-generation broodstock (F1) produced 2.00×10⁶ eggs, of which 1.90×10⁶ were fertilized. Larvae were reared at temperatures of 21.0-24.5℃. The feeding regime consisted of rotifers Artemia, and commercial pellet feed. From 5-8 days after hatching (dah), rotifers collected from high-density incubation tanks were fed to grouper larvae at a density of 10-20 individuals/mL. From 9-30 dah, rotifers collected from low-density incubation tanks were fed to larvae at a density of 10-20 individuals/mL. Newly hatched Artemia were provided from 20-24 dah and enriched Artemia from 25-50 dah. Commercial pellet feed was given starting at 13 dah. The average mouth size in hatchlings was 263.4±3.7 ㎛ at 5 dah, 406.7±21.0 ㎛ at 11 dah, and 856.9±136.6 ㎛ at 21 dah. At 1 dah, larvae had a mean total length of 1.95±0.06 mm. At 66 dah, the mean TL was 4.0 cm. The final number of surviving red spotted grouper larvae was 218,380 fishes.

• 한국축산식품학회지
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• 제36권6호
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• pp.791-798
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• 2016

Free radicals may attack cells or tissue, leading to chronic diseases, and antioxidant consumption is potentially useful for removing free radicals. Egg proteins may be used as potential sources of antioxidant considering their ability of scavenging free radicals to apply for food or cosmetics industry. In this study, we obtained a natural antioxidant protein from fertilized eggs, which was a dietary supplement in some Asian countries. Meanwhile, antioxidant activities of these proteins were evaluated using different oxidation systems. With increasing incubation time, the antioxidant activity of these proteins increased during 15 d of incubation. The samples on day 15 were performed for isolation of antioxidant protein. The protein, named P4-1 (MW, 45 kDa), was isolated and purified by consecutive chromatographic methods. P4-1 contained 17 amino acids, which was determined by liquid chromatography-mass spectrometry and Amino Acid Analyzer. Moreover, the amino acid sequence was highly similar to that of ovalbumin. Differential scanning calorimetry showed that the denaturation temperature of P4-1 was $57.16^{\circ}C$. Furthermore, P4-1 suggested high oxygen radical-absorbance activity in ${\cdot}OH$ assays, and its antioxidant activity was stable at $30-50^{\circ}C$ in acidic and neutral pH. Thus, these results revealed that P4-1 may be a potential resource as a natural antioxidant.

• Journal of Microbiology and Biotechnology
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• 제29권2호
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• pp.235-243
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• 2019

A special eosinophilic agarase exo-type ${\beta}$-agarase gene, AgaDL6, was cloned from a marine agar-degrading bacterium, Flammeovirga sp. HQM9. The gene comprised 1,383-bp nucleotides encoding a putative agarase AgaDL6 of 461 amino acids with a calculated molecular mass of 52.8 kDa. Sequence analysis revealed a ${\beta}$-agarase domain that belongs to the glycoside hydrolase family (GH) 16 and a carbohydrate-binding module (CBM_4_9) unique to agarases. AgaDL6 was heterologously expressed in Escherichia coli BL21 (DE3). Enzyme activity analysis of the purified protein showed that the optimal temperature and pH of AgaDL6 were $50^{\circ}C$ and 3.0, respectively. AgaDL6 showed thermal stability by retaining more than 98% of activity after incubation for 2 h at $50^{\circ}C$, a feature quite different from other agarases. AgaDL6 also exhibited outstanding acid stability, retaining 100% of activity after incubation for 24 h at pH 2.0 to 5.0, a property distinct from other agarases. This is the first agarase characterized to have such high acid stability. In addition, we observed no obvious stimulation or inhibition of AgaDL6 in the presence of various metal ions and denaturants. AgaDL6 is an exo-type ${\beta}$-1,4 agarase that cleaved agarose into neoagarotetraose and neoagarohexaose as the final products. These characteristics make AgaDL6 a potentially valuable enzyme in the cosmetic, food, and pharmaceutical industries.

• 한국응용과학기술학회지
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• 제32권2호
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• pp.202-214
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• 2015

Stimuli-responsive biomaterials that alter their function through sensing local molecular cues may enable technological advances in the fields of drug delivery, gene delivery, actuators, biosensors, and tissue engineering. In this research, pH-responsive hydrogel which is comprised of dimethylaminoethyl methacylate (DMAEMA) and 2-hydroxyethyl methacrylate (HEMA) was synthesized for the effective delivery of doxorubicin (Dox) to breast cancer cells. Cancer and tumor tissues show a lower extracellular pH than normal tissues. DMAEMA/HEMA hydrogels showed significant sensitivity by small pH changes and each formulation of hydrogels was examined by scanning electron microscopy, mechanical test, equilibrium mass swelling, controlled Dox release, and cytotoxicity. High swelling ratios and Dox release were obtained at low pH buffer condition, low cross-linker concentration, and high content of DMAEMA. Dox release was accelerated to 67.3% at pH 5.5 for 6-h incubation at $37^{\circ}C$, while it was limited to 13.8% at pH7.4 at the same time and temperature. Cell toxicity results to breast cancer cells indicate that pH-responsive DMAEMA/HEMA hydrogels may be used as an efficient matrix for anti-cancer drug delivery with various transporting manners. Also, pH-responsive DMAEMA/HEMA hydrogels may be useful in therapeutic treatment which is required a triggered release at low pH range such as gene delivery, ischemia, and diabetic ketoacidosis.

• Composites Research
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• 제29권4호
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• pp.132-137
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• 2016

Based on mussel-inspired polydopamine (PDA), a novel technique to fabricate carbon nanowire (CNW) arrays is presented for a possible use of porous carbon electrode in electrochemical energy storage applications. PDA can give more porosity and nitrogen-doping effect to carbon electrodes, since it has high graphitic carbon yield characteristic and rich amine functionalities. Using such outstanding properties, the applicability of PDA for electrochemical energy storage devices was investigated. To achieve this, the decoration of the CNW arrays on carbon fiber surface was performed to increase the surface area for storage of electrical charge and the chemical active sites. Here, zinc oxide (ZnO) nanowire (NW) arrays were hydrothermally grown on the carbon fiber surface and then, PDA was coated on ZnO NWs. Finally, high temperature annealing was performed to carbonize PDA coating layers. For higher energy density, manganese oxide ($MnO_x$) nanoparticles (NPs), were deposited on the carbonized PDA NW arrays. The enlarged surface area induced by carbon nanowire arrays led to a 4.7-fold enhancement in areal capacitance compared to that of bare carbon fibers. The capacitance of nanowire-decorated electrodes reached up to $105.7mF/cm^2$, which is 59 times higher than that of pristine carbon fibers.

• 생약학회지
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• 제46권4호
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• pp.279-282
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• 2015

In order to quantify compound K(CK), anticancer component of Panax ginseng C. A. Meyer, high titer rabbit polyclonal antibodies (pAbs) were raised against a conjugate of CK and bovine serum albumin coupled by a periodate oxidation method. Coating antigen (CK-OVA) was also prepared by the same method with OVA. As a result of optimization of antiserum dilution (2,000 fold), coating antigen ($25{\mu}g/ml$) and other condition (incubation time, temperature and washing method), ELISA method for the determination of CK was established. The measuring range extended from 0.5 ng/ml to 25 ng/ml of CK. The antibodies exhibited minor or even no cross reactivities with protopanaxatriol (1.56%) and other tested ginsenosides, $GRb_1$ (0.11%), $GRg_1$ (0.07%) except protopanaxadiol (87.2%) from the structural similarity. And the antibody showed good correlation (r=0.987) between the assay values obtained by this ELISA method and HPLC. Therefore, the ELISA method could be very useful tools for the determination of CK in biological fluids because of their high sensitivity and specificity.

• 한국작물학회지
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• 제29권1호
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• pp.46-49
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• 1984

맥류의 생육과 Ethylene 대사와의 관계를 측정한 몇가지 결과를 소개하면 다음과 같다. 1. 보리 엽의 Ethylene 생성량은 하위엽이 상위엽보다 많고 답압을 한 것이 하지 않은 것보다 많았다. 2. 엽신절단후 시간이 경과함에 따라 Ethylene 생성량은 증가하였는데 pot에서 보다 포장에서 재배한 것이 많았고 절엽후 Incubation의 온도가 높을수록 많았으며 주간이 야간보다 많은 경향이었다. 3. 밀의 Ethylene 생성량은 초형이 직립형인 품종 보다 포복형인 품종일수록 많고 엽록색 함량이 적은 품종보다 많은 품종일수록 증가하였다. 4. 맥류엽의 Ethylene 생성량은 복합비료보다 퇴비를 시용한 것이 많았고 담수를 한 것은 하지 않은 것보다 저하하는 경향을 보였다.

• 미생물학회지
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• 제39권3호
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• pp.155-160
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• 2003

Streptomyces viridochromogenes 야생형과 기중균사 형성 변이주들의 지질과 지방산의 조성을 배양온도와 배양시간에 따라 조사하였다. Triacylglycerol은 기중균사가 형성되는 경우에만 감소되어 기중균사 형성과 연관되어 있음을 나타내었다. 인지질의 경우에는 배양조건에 따른 특이적 변화를 볼 수 없었으나, 기중균사 형성능이 상실된 변이주 BR2가 $20^{\circ}C$에서 배양되었을 매 phosphatidylethanolamine 조성이 특이하게 높게 나타났다. 그리고 변이주 BR2는$20^{\circ}C$및 $30^{\circ}C$ 배양 시 다양한 $R_{f}$ 간의 동정되지 않은 아미노지질들을 함유하였다. 모든 균주에서 아미노지질의 일종인 ornithinolipid는 배양시간이 경과되면서 증가되는 경향을 나타내었다. 지방산조성은 온도에 민감하여 배양온도가 높을수록 불포화지방산 함량이 낮아졌다. 기중균사 형성은 직쇄형 포화지방산의 함량 감소를 수반하였으며, 기중균사 생성능이 우수한 변이주 M13은 배양초기의 직쇄형 포화지방산 함량이 가장 높았다. 변이주 BR2는 iso형 홀수탄소 지방산의 함량이 특이적으로 높았다.

• 핵의학기술
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• 제21권1호
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• pp.39-43
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• 2017

Thyroglobulin (Tg)은 갑상선에서 특이적으로 생성되는 요오드단백질로써 분화갑상선 암(Differentiate Thyroid Carcinoma, DTC) 의 생물학적 표지자이다. 본 검사실의 Tg 검사법은 반응시간이 길어 진료과와 환자들의 결과보고 요청사항을 만족시키지 못하고 있는 상황이다. 이를 만족시키기 위해 Tg 검사의 반응시간을 단축하고자 현재 사용중인 B사 Kit와 타 병원에서 사용중인 C사 Kit의 반응시간을 단축시켜 실험을 진행하였다. 본원에 의뢰된 Tg 약 65명의 환자 검체를 대상으로 B사 Kit의 반응시간을 $37^{\circ}C$ shaking 2hr/2hr, 실온 shaking 3hr/2hr, $37^{\circ}C$ shaking 1hr/1hr으로, C사 Kit의 원법인 실온 shaking 3hr법과 반응시간을 단축한 실온 shaking 2hr, $37^{\circ}C$ shaking 2hr 방법으로 단축하여 기존에 시행하던 overnight법과 상관성검사를 실시하여 비교하였다. 그 결과 여섯가지 방법 모두 1.0 ng/mL 이상의 값에서는 높은 상관관계를 보였으나 1.0 ng/mL 이하의 값에서는 다소 흔들리는 경향이 있었다. 하지만 그 중 상대적으로 안정적이었던 B사 Kit $37^{\circ}C$ shaking 1hr/1hr 법과 C사 Kit 실온 shaking 2hr 법에 대해 다른 조건들을 재설정하여 적합한 방법을 찾는다면 진료과와 환자들의 요구 사항 충족은 물론, EIA법과의 경쟁력 면에서도 유용할 것이다.