• 한국축산식품학회지
• /
• 제22권4호
• /
• pp.343-347
• /
• 2002

TGF-$\beta$l은 여러 가지 생리활성 기능을 가지고 있기에 기능성식품 및 의약품 소재로 이용될 수 있다. 본 연구에서는 bovine colostrum milk로부터 TGF-$\beta$l을 분리 정제하기 위해 Cel-filtration chromatography, AF-heparin column chromatography 및 AF-heparin column rechromatography를 수행하여 TGF-$\beta$l을 정제하였다. 정제된 TGF-$\beta$1은 비환원조건하에서 전기영동을 수행하여 표준 TGF-$\beta$l과 같은 위치에 단일 band가 나타남으로 TGF-$\beta$l임을 확인하였다. 또한 환원 조건하에서 Western blot을 수행한 결과 TGF-$\beta$l 단일클론 항체와 결합하는 monomer 형태의 밴드를 확인하였다. 정제 TGF-$\beta$1의 회수율은 21%였다.

• 한국미생물·생명공학회지
• /
• 제31권1호
• /
• pp.51-56
• /
• 2003

안정성이 확보된 식품에서 ACE저해 활성 물질을 검색하는 연구의 일환으로, 옥수수 글루텐을 Flavourzyme, Pescalase, 그리고 Thermolysine/Pescalase 등으로 가수분해하여 얻은 가수 분해물로부터 ACE 활성 저해 펩타이드를 다음과 같은 과정으로 분리, 정제하였다. 10% 에탄올로 평형화된 ODS chromatography를 이용 단백질 분획들을 얻고, Bio-Gel P-2 column과 reverse phase HPLC를 통해 5개의 ACE 저해 펩타이드를 분리, 정제하였다. 그 아미노산 서열은 LPF($IC_{50}$ = 40 $\mu$M), GPP($IC_{50}$ = 17.6 $\mu$M), PNPY($IC_{50}$ = 30.7 $\mu$M), SPPPFYL($IC_{50}$/ = 63 $\mu$M), and SQPP($IC_{50}$ = 17.2 $\mu$M)로 밝혀졌다. 이 펩타이드들은 경구투석 시 가수분해 효소에 대응하여 체내에서 안정성이 뛰어나고, 소장에서도 쉽게 흡수될 것으로 사료되어 상시 섭취하는 식품이나 음료에 첨가하여 이용한다면 그 유용성이 기대된다.

• 한국환경보건학회지
• /
• 제31권4호
• /
• pp.309-315
• /
• 2005

Volatile organic compounds (VOCs) and odor materials are major sources of air pollution in Ulsan city, where much chemical plants are located. Therefore, it is necessary to develop a new reactor which can remove VOCs and odor materials effectively and be equipped at the end of pipe easily. A modified reactor (bioscrubber trickling filter, BSTF), which have both characteristics of biofilter and bioscrubber, was developed and tested on its reactivity with several VOCs using two types of media, fiber and activated carbon 4- ceramic(A/C). It was observed that the removal efficiencies of several types of VOCs such as acetaldehyde, ethylalcohol, butanol, diethylamine and triethylamine were up to $95\%$ when they had about 100 ppm of initial concentration and 80 seconds of residence time. Good attachment of microorganisms to both media, where it is thought the reaction efficiency can be determined according to the amount of microorganisms attachment, observed with scanning electron microscopy(SEM). Initial pressure drops of the packed bed with both media were 229 $mmH_2O/m$ at A/C column and 670 $mmH_2O/m$, respectively. However, maximum pressure drop of fiber column during the operation was over 1,647 $mmH_2O/m$. Therefore, it was thought that the fiber material would not suitable to use in the local plant as a packed bed media.

• Applied Biological Chemistry
• /
• 제41권7호
• /
• pp.522-526
• /
• 1998

콩 유용성분 탐색의 일환으로 그리고 향후 콩 ferritin 항체 및 유전자 확보를 목표로 발아된 콩으로부터 ferritin을 분리 정제하여 그 몇가지 특성을 조사하였다. 72시간 발아된 콩으로부터 ammonium sulfate 침전(0.55 saturation), DEAE-cellulose, Sephacryl S-300, Bio-Scale Q2 column chromatographies를 통하여 ferritin을 분리하였다. 정제된 콩 ferritin은 SDS-PAGE 분석에서 21 kDa의 크기를 나타냈으며, Sephacry S-300을 통한 겔거르기 chromatography와 non-denaturing 폴리아크릴아마이드 전기영동 분석에서 $510{\sim}560\;kDa$의 크기로 측정 되었다. 또한, immunodiffusion test에서 anti-soybean ferritin antiserum과 상호 반응하였다. 원자흡광광도계와 표준 철 용액을 이용한 정제된 콩 ferritin 중의 철 함유량은 833 mol Fe/mol protein 이었으며, 이는 호박씨로부터 분리한 ferritin보다 31배 더 많은 양의 철 함유량 이었다. 정제된 콩 ferritin중의 철은 horse spleen ferritin 중의 철과 유사하게 iron staining 되었다.

• 한국식품과학회지
• /
• 제28권1호
• /
• pp.99-104
• /
• 1996

탄수화물을 갖고 있지 않은 caseinomacropeptide (CMP)를 감성유청분말로부터 12% TCA법에 의해 분리한 결과 100 g의 감성유청분말로 부터 2.7 g을 얻을 수 있었다. 분리된 CMP는 전기영동과 아미노산 조성을 분석한 결과 매우 순도가 높은 CMP로 나타나 12% TCA침전법은 감성유청분말로 부터 CMP를 분리하는 효과적인 방법임을 알 수 있었다. 트립신을 pore glass beads에 carbodiimide (EDC) 방법으로 amide결합에 의해 고정화시켰으며 고정화된 트립신의 양은 1 g glass beads당 20 mg이었다. 수용성 트립신에 의한 CMP 가수분해는 24시간이 지나면 거의 완료되었으며 고정화 트립신은 24시간이 지난 후에도 가수분해가 진행되고 있었다. 가수분해물을 Bio-Gel P4 column에 의해 분획한 후 혈소판 응집 저해작용을 측정한 결과 고정화 트립신에 의해 24시간 가수분해시킨 CMP 분해물이 가장 높은 혈소판 응집 억제능을 보였다.

• 한국식품과학회지
• /
• 제28권1호
• /
• pp.184-189
• /
• 1996

냉이(Capsella bursa-pastoris)로부터 산소독성에 영향을 미치는 superoxide anion radical에 대하여 소거작용을 하는 물질을 분리하고 이 물질의 이화학적 성질을 검토하였다. 냉이를 에탄올로 추출한 후 용매분획과 각종 column chromatographies (Amberlite XAD-2, Sephadex LH-20, Bio gel P-2, ODS)를 통해 정제하였고, 최종적으로 FPLC (Fast protein liquid chromatography)를 사용하여 활성의 단일물질을 얻었다. 냉이 에탄올추출물 100 g으로부터 0.25 g의 정제물질을 얻었으며, superoxide anion radical을 50% 소거하는 농도$(IC_{50})$는 $0.58\;{\mu}g$이었다. 이 물질에 대해 각종 이화학적 성질을 검토한 결과 phenol성 화합물의 배당체로 추정하였다.

• Journal of Biomedical and Translational Research
• /
• 제19권4호
• /
• pp.130-139
• /
• 2018

This study aimed to analyze a high-performance liquid chromatography (HPLC) separation using a pentafluorophenyl column of parent drug hydroxychloroquine (HCQ) and its active metabolite, desethylhydroxchloroquine (DHCQ) applying to determine bioequivalence of two different formulations administered to patients. A rapid, simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for bioanalysis of HCQ and its metabolite DHCQ in human whole blood using deuterium derivative $hydroxychloroquine-D_4$ as an internal standard (IS). A triple-quadrupole mass spectrometer was operated using electrospray ionization in multiple reaction monitoring (MRM) mode. Sample preparation involves a two-step precipitation of protein techniques. The removed protein blood samples were chromatographed on a pentafluorophenyl (PFP) column ($50mm{\times}4.6mm$, $2.6{\mu}m$) with a mobile phase (ammonium formate solution containing dilute formic acid) in an isocratic mode at a flow rate of 0.45 mL/min. The standard curves were found to be linear in the range of 2 - 500 ng/mL for HCQ; 2 - 2,000 ng/mL for DHCQ in spite of lacking a highly sensitive MS spectrometry system. Results of intra- and inter-day precision and accuracy were within acceptable limits. A run time of 2.2 min for HCQ and 2.03 min for DHCQ in blood sample facilitated the analysis of more than 300 human whole blood samples per day. Taken together, we concluded that the assay developed herein represents a highly qualified technology for the quantification of HCQ in human whole blood for a parallel design bioequivalence study in a healthy male.

• Journal of Microbiology and Biotechnology
• /
• 제17권4호
• /
• pp.638-643
• /
• 2007

The full encoding sequence for human type II hexokinase (HXK II) was cloned into the E. coli expression vector pET 21b and expressed as a C-terminally hexahistidine-tagged protein in the BL2l (DE3) strain. The IPTG-induced HXK II approximately accounted for 17% of the total E. coli proteins, and 81% of HXK $II_{6{\times}His}$ existed in inclusion bodies. To improve the production of soluble recombinant HXK II protein, in the functionally active form, we used low temperature, and the osmotic stress expression method. When expressed at $18^{\circ}C$, about 83% of HXK $II_{6{\times}His}$ existed in the soluble fraction, which amounted to a 4.1-fold yield over that expressed at $37^{\circ}C$. The soluble form of HXK $II_{6{\times}His}$ was also highly produced in the presence of 1M sorbitol under the standard condition $(37^{\circ}C)$, which indicated that temperature downshift and low water potentials were required to improve the yield of active recombinant HXK II protein. The expressed protein was purified by metal chelate affinity chromatography performed in an IDA Excellose column charged with $Ni^{2+}$ ions, resulting in about 40mg recombinant HXK II protein obtained with purity over 89% from 51 of E. coli culture. The identity of HXK $II_{6{\times}His}$ was confirmed by Western blotting analysis. Taken together, using the stress-governed expression described in this study, human active HXK II can be purified in sufficient amounts for biochemical and biomedical studies.

• 한국토양비료학회지
• /
• 제47권2호
• /
• pp.113-119
• /
• 2014

Arsenic and its compounds which is one of the most toxic elements that can be found naturally on earth in small concentrations are used in the production of pesticides, herbicides, and insecticides. Most arsenic that cannot be mobilized easily when it is immobile is also found in conjunction with sulfur in minerals such as arsenopyrite (AsFeS), realgar, orpiment and enargite. In this investigation we observed the leaching of arsenic in soils amended with several levels of gravel size of arsenopyrite collected from a road construction site. Soil and gravel size of arsenopyrite were characterized by chemical and mineralogical analyses. Results of XRF analysis of arsenopyrite indicated that the proportion of arsenate was 0.075% (wt $wt^{-1}$) while the maximum amount of arsenic in soil samples was 251.3 mg $kg^{-1}$. Cumulative amounts of effluent collected from the bottom of the soil column for different mixing rate of the gravel were gradually increased where proportion of the gravel mixed was greater than 70% whereas the effluent was stabilized to the maximum after approximately 45 pore volumes of effluent or greater were collected. The arsenic in the effluent was recovered from the soil columns in which the proportion of arsenopyrite gravel was 60% or greater. The total amount of arsenic recovered as effluent was increased with increasing proportion of gravel in a soil, indicating that the arsenic in the effluent was closely related with gravel fraction of arsenopyrite.

• 한국축산식품학회지
• /
• 제29권2호
• /
• pp.157-163
• /
• 2009

This study was conducted for the isolation, purification, and characterization of a protease from Korean pear, to see its proteolytic activity on chicken actomyosin and to find the optimum pH and temperature of activity on chicken actomyosin. The protease was isolated from crude extract of Korean pear by ammonium sulfate precipitation. Further purification was done by DEAE-Sepharose ion-exchange chromatography, Mono-Q and Mini-Q column chromatography. The purified enzyme gave a single protein band on SDS polyacrylamide gel electrophoresis and the molecular weight was found to be 38 kDa. The specific activity of purified enzyme was 34,907 unit/mg with 25 fold purification and the yield was 2%. The purified enzyme incubated with chicken actomyosin showed high activity. The optimum pH and temperature for enzyme activity on chicken actomyosin were 6.5 and $70^{\circ}C$, respectively. A protease was purified from Korean pear for the first time and characterized. It was found to be promising for meat tenderization.