• 멤브레인
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• 제31권2호
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• pp.145-152
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• 2021

현재 바이오 분야에서 분리에 대한 수요가 급증함에 따라, 투과율 및 결합능 측면에서 높은 성능을 띠는 막크로마토그래피가 수지 크로마토그래피의 대체 분리 공정으로 부상하고 있다. 실증을 기반으로 하여 막 소재가 결정되는 기존 분리막 공정과 달리, 막크로마토그래피의 경우 분리하고자 하는 목표 물질에 적합한 분리 메커니즘 이해 그리고 이를 기반한 공정 설계가 필요하다. 본 논문에서는 생특이성을 활용하여 선택적으로 거대 분자를 포집하는 친화성 작용, 전하를 활용하여 생분자와 결합하는 이온 교환 작용 그리고 소수성을 활용하여 생분자와 결합하는 소수성 작용과 같은 막크로마토그래피 주요 분리 메커니즘들에 대해 다루고자 한다. 또한 본 논문에서는 단계적 측면에서 또는 소재 측면에 막크로마토그래피 기술설계 시 고려해야할 변인들에 대해서 다루고자 한다.

• 한국미생물·생명공학회지
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• 제31권2호
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• pp.201-204
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• 2003

Glycopeptide antibiotics, teicoplanin was purified from a mutant strain of Actinoplanes teichomyceticus ATCC31121, A. teichomyceticus MSL2211. We developed a simple procedure to separate and purify the teicoplanin from the fermentation broth. Teicoplanin was purified by two-step purification system, hydrophobic adsorption and sugar affinity chromatography in combination with HPLC analysis based on the properties of hydrophobic acyl chain and sugar moiety in teicoplanin. Teicoplanin was separated from the culture broth by Diaion HP-20 and further purified by concanavalin A affinity column chromatography. As an adsorbent resin, Diaion HP-20 in broth eliminated toxic effects on growth, reduced feedback repression of teicoplanin production, and assisted In rapid recovery of teicoplanin. The teicoplanin displayed the final yield of 80% and 95% of purity.

• 한국미생물·생명공학회지
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• 제26권5호
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• pp.413-419
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• 1998

Acetaminophen 자화성 Pseudomonas sp.에 존재하는 aryl acylamidase[EC 3.5.1.13]는 ammonium sulfate fractionation, DEAE-Sephacel anion exchange chromatography, Phenyl-Sepharose CL-4B hydrophobic interaction chroamtography 및 Sephadex G-100 gel-permeation chromatography를 통해 순수 정제되었다. 정제된 효소는 SDS-PAGE상에서 분자량을 측정한 결과 56 kDa, Sephadex G-100 gel-permeation chromatography로 측정한 결과 57 kDa이었으며, 따라서 단일한 subunit로 구성된 효소이었다. 정제된 효소의 최대 활성을 위한 pH와 온도는 각각 pH 10.5와 4$0^{\circ}C$이였다. 5$0^{\circ}C$에서 30분간 처리시 34%의 잔존활성을 나타내었다. Acetaminophen과 4'-nitroacetanilide쉐 대한 Km값은 각각 0.10 mM과 0.11 mM 이었다.

• Bulletin of the Korean Chemical Society
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• 제14권4호
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• pp.515-519
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• 1993

The retention behavior of proteins was investigated by using hydrophobic interaction chromatography (HIC), comparing to the results obtained in reversed-phase chromatography (RPC) described in the previous paper. A SynChropak propyl column was employed with 0.05 M phosphate buffer (pH 7.0) containing sodium sulfate. Conformational changes were recognized by examining Z values as a function of sodium sulfate concentration over a range of temperature between 5 and 65$^{\circ}C$. Z values did not change significantly at the range of the temperature showing the consistent ${\Delta}H^{\circ}$ and ${\Delta}S^{\circ}$ values. The sign and the magnitude of ${\Delta}H^{\circ}$ and ${\Delta}S^{\circ}$ of proteins in HIC were compared with those obtained in RPC. The signs of ${\Delta}H^{\circ}$ and ${\Delta}S^{\circ}$ of proteins in HIC were all positive, while those of proteins in RPC were all negative. These results suggested that the retention of proteins in HIC and in RPC were entropy-driven and enthalpy-driven process, respectively. From the two different investigations, it was concluded that the retention mechanism of RPC and HIC was based on the same fundamental principle in which separation is dependent on hydrophobicity, but the retention behavior of the proteins in HIC is clearly different from that observed in RPC.

• 미생물학회지
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• 제44권2호
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• pp.116-121
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• 2008

Haloperoxidase를 생산하는 미생물을 분리하기 위하여 국내 연근해와 남북극 등의 해양시료에서 분리된 방선균 균주를 대상으로 탐색을 수행하여 남해 백도 해조류 추출물로부터 분리된 한 종류의 방선균(#1460)에서 높은 haloperoxidase 활성이 확인되었다. 본 균주의 생리.생화학적 특성은 Streptomyces 속과 유사하며 생산되는 haloperoxidase는 세포 조 추출물로부터 ammonium sulfate precipitation, High-Q column chromatography, gel permeation chromatography, Hydroxyapetite chromatography 그리고 hydrophobic interaction chromatography를 통하여 42%의 수율과 purification fold 70으로 정제하였다. 본 효소의 최적 반응 pH는 7이고 pH 8에서 더 높은 안정성을 보여 $60^{\circ}C$에서 1시간 반응에 효소활성의 50%가 생존한다. 또 cyanide와 azide 이온에 의해 강한 저해현상을 보인다.

• 약학회지
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• 제35권6호
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• pp.461-465
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• 1991

Silica-based gel filtration chromatography has been used to characterize molecular weight of proteins. However, the molecular weight measured by this method was distorted by protein-silica interactions like hydrophobic and electrostatic forces. Therefore, we characterized protein-silica interaction using two forms of phytochrome (124 kDa) having different hydrophobicity and surface charge. PH and ionic strength affected the retention time of phytochrome suggesting that electrostatic force is the major interaction between protein and silica surface.

• Bulletin of the Korean Chemical Society
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• 제18권6호
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• pp.648-653
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• 1997

Acetolactate synthase was purified from Escherichia coli MF2000/pTATX containing Arabidopsis thaliana acetolactate synthase gene. Purification steps included DEAE cellulose ion exchange column chromatography, phenyl sepharose hydrophobic column chromatography, hydroxylapatite affinity column chromatography, and Mono-Q HPLC. Molecular weight was estimated to be ∼65 KDa and purification fold was 109 times. The enzyme showed a pH optimum of 7 and the $K_M$ value was 5.9 mM. The purified enzyme was not inhibited by any of the end products, valine, leucine, and isoleucine.

• Macromolecular Research
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• 제16권5호
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• pp.418-423
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• 2008

Poly[(R)-3-hydroxy butyrate] (PHB) and methoxy poly(ethylene glycol) (mPEG) were conjugated by the transesterification reaction with tin(II)-ethylhexanoate (Sn(Oct)-II) as a catalyst. Hydrophobic PHB and hydrophilic mPEG formed an amphiphilic block copolymer which was formed with the self-assembled polymeric micelle in aqueous solution. In this study, we tried to determine the optimum ratio of hydrophobic/hydrophilic segments for controlled drug delivery. The particle size and shape of the polymeric micelle were measured by atomic force microscopy (AFM) and transmission electron microscopy (TEM). Their size were 61-102 nm with various block ratios. Griseofulvin was loaded in the polymeric micelle as a hydrophobic model drug. The loading efficiency and release profile were measured by high performance liquid chromatography (HPLC). The model drug in our system was constantly released for 48 h.

• 한국전기전자재료학회:학술대회논문집
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• 한국전기전자재료학회 2004년도 하계학술대회 논문집 Vol.5 No.1
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• pp.503-506
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• 2004

In this paper we investigated the changes of surface hydrophobic characteristics on silicone rubbers by corona discharge treatment and also investigated the distribution and the behavior of low molecular weight(LMW) silicone fluid which was extracted by solvent-extraction with gel permeation chromatography(GPC). It was shown that contact angle was $110.5^{\circ}$ on initial sample but contact angle was approximately decreased to $10^{\circ}$ after 45 minutes. However the surface hydrophobic characteristic on silicone rubbers which were removed from corona discharge was recovered within 5 hours. It was shown that corona discharge insured the increase of diffusible LMW chains, which could lead to recover the surface hydrophobicity. The surface hydrophobic characteristics on silicone rubbers and the recovery mechanism based on our results were discussed.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 1986년도 추계학술대회
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• pp.515.1-515
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• 1986

Thermostable tryptophanase was extracted from a thermophilie bacterium, strain T which was absolutely symbiotic with strain 5. The enzyme was purified 14.7 fold with 5.8% yield by chromatographies using ion exchange, gel filtration, and hydrophobic interaction columns, followed by high performance liquid chromatography on hydroxyapatite column. The purified enzyme has a molecular weight of approximately 210,000 estimated by gel filtration column chromatography, and the molecular weight of subunit was determined by SDS polyacrylamide gel electrophoresis to be 46,000, which indicates that the native enzyme is made of four homologous subunits. The tryptophanase was stable at 65o0 and the optimum temperature for the enzyme activity for 20 min reaction was 70$^{\circ}C$. The purified enzyme activity for 20 min ieaction was 70$^{\circ}C$. The purified enzyme catalyzed the degradation of L-tryptophan into indole, pyruvate and ammonia in the presence of pyridoxal phosphate. 5-Hydroxy-Ltryptophan, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-L-cysteine, 5-methyl-DL-tryptophan, L-cysteine, S-methyl-Lcysteine, and L-serine were also used as substrates to form pyruvate. The amino acid composition of the tryptophanase was determined, and found to contain a high percentage of hydrophobic amino acids, especially in the proline content, which was much higher than that of Escherichia coli tryptophanase. In addition, the 35N-terminal amino acid sequence of the tryptophanase was completely different from that of E. coli tryptophanase.