• KSBB Journal
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• 제14권3호
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• pp.371-376
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• 1999

제조용 액체 크로마토그래피를 전산모사하는 것은 최적의 분리와 조건을 위해 필수적이다. 제조용 액체 크로마토그래피에서의 단일 성분과 2성분 용출 피크의 거동을 설명하는 수학적 모델을 equilibrium-dispersive model을 이용해 계산을 수행하였다. 모델의 전산모사를 통해 주입되는 시료부피, 시료속의 용질의 농도, 유속, 컬럼길이가 용출되는 피크에 미치는 영향을 확인해 보았다. Equilibrium-dispersive model로 전산모사된 data를 이용하여 단일 성분 시료 주입에 대해 주입 농도와 유속에 대한 영향을 알아본 결과 주입 농도를 증가시킬수록 용출이 일어나는 시점이 앞당겨져 평균 체류 시간이 감소하고 날카로운 피크를 얻을 수 있다. 유속을 증가시키면 시료의 용출 시점이 앞당겨지고, 고정상과의 충분한 상호 작용을 일으키기 전에 용출되기 때문에 피크가 날카로와 진다. 2성분 시료 주입에 대해 분리도값들을 계산한 결과 두번째로 용출되는 시료의 농도를 증가시킬수록 이 시료성분이 더 빨리 용출되므로 분리도값이 감소함을 알 수 있다. 칼럼길이가 분리도에 미치는 영향을 보면 칼럼길이를 길게 할수록 시료성분이 고정상과의 완전한 상호작용으로 분리가 잘되고, 시료의 주입량을 증가시키면 두 성분간의 용출 피크의 간격이 좁아져 분리도 값이 감소함을 알 수 있다. 이 내용을 바탕으로 다성분 분리에 대한 simulation 연구로 확장할 수 있다.

• Journal of Microbiology and Biotechnology
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• 제19권4호
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• pp.416-423
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• 2009

Hepatitis B core antigen(HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus(HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the "hold-up" period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test.

• 한국동물학회지
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• 제37권1호
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• pp.66-75
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• 1994

Juvenile hormone binding protein was identified in the hemolymph of fifth instar larvae and purified using column chromatography. Hemolymph was mixed with [3H] JH-III and electrophoresed on 691 NON-SDS gel, indicating that radioactivity peak appears at Rf value of 0.55. Gel filtration showed two radioactivity peaks equivalent to bound and free [3H]JH-III, respectively. JHBP was purified from hemolymph through gel filtration (Sephadex G-100), anion exchange chromatosraphv (DEAE Sepharose CL-6B), chromatofocusing chromatographv (PBE 94) and preparative electrophoresis.

• Journal of Microbiology
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• 제37권1호
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• pp.21-26
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• 1999

Two novel calcium-binding proteins, named CAB-I and CAB-II, have been isolated from Streptomyces coelicolor. Purification of the calcium-binding proteins involved heat treatment, fractionation with ammonium sulfate, acid treatment, anion exchange and hydrophobic interaction column chromatography, FPLC gel filtration, and preparative isoelectric focusing. A chelex competitive assay and 45Ca autoradiography verified the calcium-binding ability of the proteins. The major band CAB-II has an apparent molecular weight of 26,000 determined by SDS-polyacrylamide gel electrophoresis and 340,000 determined by gel filtration. The isoelectric point of this molecule showed the acidic nature of the molecule. N-terminal amino acid sequence analysis shows homology to rat Ca2+/calmodulin-dependent protein kinase-II (CAB-II) and yeast phosphoprotein phosphatase (CAB-I).

• Animal cells and systems
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• 제3권2호
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• pp.173-180
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• 1999

A blue biliprotein, insecticyanin (INS), has been purified from the last instar larval hemolymph of Agrius convolvuli by ultracentrifugation, Sephadex G-100 gel permeation chromatography, and preparative electrophoresis. The molecular mass of INS was estimated to be 26 kDa and the N-terminal sequence of INS revealed high similarity to that of Manduca sexta. Results of Western blotting and autoradiography indicated that INS is synthesized by the epidermis and released into the hemolymph. In contrast to the INS reported in other insects, Agrius convolvuli INS contained a small amount of lipid, predominately consisting of triacylglycerol. Subcellular localization of INS was determined using protein-A gold particles linked to secondary antibodies (anti-rabbit Ig). INS was heavily accumulated in the cytoplasmic inclusion body (CIB). CIBs showed a variety of shapes from rod to globule and generally surrounded the nucleus. They were mostly located near the basement membrane and especially abundant in the intersegmental membrane.

• 한국미생물·생명공학회지
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• 제21권1호
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• pp.18-22
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• 1993

Oligofructo당 생산에 이용될 수있는 endoinulinase 분석방법은 그 균주가 endo- 및 exoinulinase를 함께 내는 경우, 일반적인 환원당 분석법으로는 정량하기가 어려워진다. 이 실험에서는 endoinulinase만을 선택적으로 정량하기 위한 하나의 방법으로써 항체 분석법을 이용하였다. Aspergillus niger ATCC 16882 조효소액을 CM-DEAE ion exchange chromatography, pI 2.5-5에서 preparative isoelectric focusing, HPLC gel filtration을 통해 순수하게 endoinulinase에 대한 항체를 얻었다.DEAE-ion exchange 및 protein A에서 정제된 이항체는 immunoassay 한 결과, exoinulinase 가 아닌 endoinulinase와 만 특이하게 반응하였고, immuno affinity chromatography 결과들은 배양액 중의 다른 단백질과 반응하지 않음이 확인되었다. 이눌린을 유일한 탄소원으로 한 배지에서 자란 1200여개의 야생균주들로부터 배양 특성이 우수한 균주를 1차로 선별하고 이 균주들의 endoinulinase 함량을 rocket immunoassay를 통하여 조사하였다. 이 중 1개의 균주는 Novozyme의 ATCC 1688와 비교할만한 정도의 endoinulinase를 배양액 중에 분비함을 확임할 수 있었다.

• Journal of Microbiology
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• 제33권2호
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• pp.103-108
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• 1995

Streptomyces fradiae protease inhibitor (SFI) was purified effectively by preparative isoelectric focusing and hydroxyapatite chromatography. The molecular weight of SFI was estimated to be 1.7 kDa by SDS-PAGE and 1.8 kDa by molecular sieving HPLC. One hundred and sixty amino acid residues were determined from which molecular weight of SFI was calculated to be 17.054 Da and carbohydrate residue was not detected. SFI was calculated to be 17,064 Da and carbohydrate residue was not detected. SFI was a monomeric protein with two reactive sits, of which isoelectric point was pH 4.1. N-terminal amino acid sequence of SFI had homology with SSI (Streptomyces subsilisin inhibitor) and other protease inhibitors produced by Streptomyces.

• Journal of Dairy Science and Biotechnology
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• 제39권1호
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• pp.36-50
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• 2021

This study aimed to evaluate the biological potential of functional food, i.e., specific peptides obtained from the hydrolysis of milk protein, by assessing their antioxidant and antibacterial properties. For the preparation of casein hydrolysates, commercial enzymes were added to 10% casein solution in a 1:200 (w/v) ratio, and samples were collected each hour. Based on the assessment of the degree of hydrolysis (DH) of casein hydrolysates, it was observed that the concentration of all enzymatic hydrolysates increased rapidly from 30 to 40 minutes. However, no change was observed in their concentrations after 150 minutes. Protamex® and Neutrase® exhibited the highest DH when compared to other enzymes. Furthermore, SDS-PAGE was performed for analyzing the proteolytic pattern of each enzyme, except for Flavourzyme®, and peptides in the size range of 20-25 kDa were identified. Subsequently, peptides produced by two enzymes were isolated using a preparative liquid chromatography system. Overall, NF3, NF4, PF5, and PF6 showed higher antioxidant potential than other peptide fractions. Moreover, NF7 and PF3 exhibited the highest antibacterial activity. In this study, we evaluated the biological potential of novel casein-derived peptides that may find application in the food and healthcare industry.

• 생약학회지
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• 제45권3호
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• pp.209-213
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• 2014

The anti-melanogenic substance was isolated from the root of Angelica gigas Nakai by silica gel column chromatography, preparative HPLC and TLC. As a result of the structure analysis by mass, $^1H$-NMR, and $^{13}C$-NMR spectrometry, the compound was identified as demethylsuberosin. Demethylsuberosin reduced melanin contents of B16F1 melanoma cells in a dose-dependent manner and decreased to about 74% at a concentration $5{\mu}g/ml$. Demethylsuberosin inhibited the expression in microphthalmia associated transcription factor (MITF), tyrosinase, tyrosinase related protein-1 (TRP-1), and tyrosinase related protein-2 (TRP-2) in melanocytes. These results suggest that the whitening activity of demethylsuberosin may be due to the inhibition of the melanin synthesis by down-regulation of MITF, tyrosinase, TRP-1 and TRP-2 expression. Thus, our results provide evidence that demethylsuberosin might be useful as a potential skin-whitening agent.

• 한국식품영양과학회지
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• 제15권3호
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• pp.276-285
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• 1986

능이 [Sarcodon aspratus (Berk.) S. Ito]로부터 단백질가수분해(蛋白質加水分解) 효소(酵素)를 분리 정제하여 그 특성을 조사하였다. 본 효소는 Tri-acryl CM-cellulose, Ultrogel AcA 54, Hydroxy apatite column chromatography와 preparative isoelectric focusing 방법으로 순차 정제되었고, 정제된 효소는 PAGE상에서 단일 band를 나타내었으며 활성(活性)은 62.5O.D/mg.protein으로 조효소(粗酵素)보다 8.01배 증가하였다. 작용최적(作用最適) pH는 10.1로 alkaline protease였으며 작용최적(作用最適) 온도(溫度)는 $57^{\circ}C$부근이었다. $50^{\circ}C$ 이하의 온도(溫度)와 pH $4.0{\sim}10.8$ 범위에서 안정(安定)하였으나, $60^{\circ}C$에서 30분후 26%, $65^{\circ}C$에서는 65%가 실활(失活)되었다. $Mn^{++}$은 효소의 활성(活性)을 증가시켰으나 $Hg^{++}$와 $Cu^{++}$는 현저히 저해시켰다. SDS-PAGE에 의해 분자량(分子量)은 30,100으로 측정되었고, monomer임이 확인되었다. 등전점(等電点)은 9.80이었다.