• Molecules and Cells
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• 제39권3호
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• pp.211-216
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• 2016

CYP107W1 from Streptomyces avermitilis is a cytochrome P450 enzyme involved in the biosynthesis of macrolide oligomycin A. A previous study reported that CYP107W1 regioselectively hydroxylated C12 of oligomycin C to produce oligomycin A, and the crystal structure of ligand free CYP107W1 was determined. Here, we analyzed the structural properties of the CYP107W1-oligomycin A complex and characterized the functional role of the Trp178 residue in CYP107W1. The crystal structure of the CYP107W1 complex with oligomycin A was determined at a resolution of $2.6{\AA}$. Oligomycin A is bound in the substrate access channel on the upper side of the prosthetic heme mainly by hydrophobic interactions. In particular, the Trp178 residue in the active site intercalates into the large macrolide ring, thereby guiding the substrate into the correct binding orientation for a productive P450 reaction. A Trp178 to Gly mutation resulted in the distortion of binding titration spectra with oligomycin A, whereas binding spectra with azoles were not affected. The Gly178 mutant's catalytic turnover number for the 12-hydroxylation reaction of oligomycin C was highly reduced. These results indicate that Trp178, located in the open pocket of the active site, may be a critical residue for the productive binding conformation of large macrolide substrates.

• 한국막학회:학술대회논문집
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• 한국막학회 1998년도 춘계 총회 및 학술발표회
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• pp.99-101
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• 1998

1. Introduction : The conventional grafting polymerization technique requires chemically reactive groups on the surface as well as on the polymer chains. For this reason, a series of prefunctionalization steps are necessary for covalent grafting. The surface prefunctionalizational technique for grafting can be used to ionization radiation, UV, plasma, ion beam or chemical initiators. Of these techniques, radiation method is one of the useful methods because of uniform and rapid creation of active radical sites without catalytic contamination in grafted samples. If the diffusion of monomer into polymer is large enough to come to the inside of polymer substrate, a homogeneous and uniform grafting reaction can be carried out throughout the whole polymer substrate. Radiation-induced grafting method may attach specific functional moieties to a polymeric substrate, such as preirradiation and simultaneous irradiation. The former is irradiated at backbone polymer in vacuum or nitrogen gas and air, and then subsequent monomer grafting by trapped or peroxy radicals, while the latter is irradiated at backbone polymer in the presence of the monomer. Therefore, radiation-induced polymerization can be used to modification of the chemical and physical properties of the polymeric materials and has attracted considerable interest because it imparts desirable properties such as blood compatibility. membrane quality, ion excahnge, dyeability, protein adsorption, and immobilization of bioactive materials. Synthesizing biocompatible materials by radiation method such as preirradiation or simultaneous irradiation has often used $\gamma$-rays to graft hydrophilic monomers onto hydrophobic polymer substrates. In this work, in attempt to produce surfaces that show low levels of anti-fouling of bovine serum albumin(BSA) solutions, hydroxyethyl methacrylate(HEMA) was grafted polypropylene membrane surfaces by preirradiation technique. The anti-fouling effect of the polypropylene membrane after grafting was examined by permeation BSA solution.

• Korean Chemical Engineering Research
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• 제56권2호
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• pp.162-168
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• 2018

유기 및 무기 폴리실라잔을 우레탄 아크릴레이트와 혼합시킴에 따라 UV 경화형 하드 코팅 용액을 제조하였다. 이용액을 polymethylmethacrylate (PMMA) 시트에 흐름 코팅한 후 UV 경화시킴에 의해 UV 경화형 하드 코팅 도막을 제조하였다. 이 과정 중 폴리실라잔의 종류 및 첨가량을 변화시켜 코팅 도막의 물성에 미치는 영향을 살펴보았다. 그 결과 유기 폴리실라잔의 경우 $95^{\circ}$의 수접촉각을 보여 높은 소수성을 나타내었으며, 7H의 연필경도와 92%의 가시광선 투과율을 보였다. 반면에 무기 폴리실라잔을 코팅한 도막은 8H의 높은 연필경도와 기재와의 우수한 접착력 및 $82^{\circ}$의 수접촉각을 나타내었다.

• 마이크로전자및패키징학회지
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• 제17권1호
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• pp.75-80
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• 2010

다공성 탄소전극기지 위의 무전해 니켈도금에 관한 연구를 하였다. 다공성 탄소전극기지로는 다공도가 20 ${\mu}m$ 이상인 것과 16~20 ${\mu}m$ 인 것을 사용하였다. 소수성인 탄소 표면은 $60^{\circ}C$ 이상의 암모니아 용액에 침적함으로써 그 표면 성질이 친수성으로 변화 되었고, 40분 이상 침적 시 접촉각이 $20^{\circ}$ 이하까지 측정 되었다. 도금욕의 pH가 증가됨에 따라 탄소기지 위에 도금된 니켈 도금층의 인의 석출량은 감소하였으며 니켈 도금층이 결정질 구조를 갖는 현상이 관찰되었다. 도금층의 두께는 pH가 증가함에 따라 증가하였다. 활성화 처리를 위한 $PdCl_2$의 농도에 따른 도금층의 두께 변화는 없었으나, 도금에 필요한 $PdCl_2$의 최소농도는 5 ppm 이상인 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제18권5호
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• pp.852-858
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• 2008

An extracellular protease (Mc1) was isolated from the nematode-trapping fungus Monacrosporium cystosporium by gel filtration, anion-exchange, and hydrophobic interaction chromatographies. This protease had a molecular mass of approximately 38 kDa and displayed an optimal activity at pH 7-9 and $56^{\circ}C$ (over 30 min). Its proteolytic activity was highly sensitive to the serine protease inhibitor PMSF (phenylmethylsulfonylfluoride, 0.1 mM), indicating that it belonged to the serine-type peptidase group. The Michaelis constant ($K_m$) and $V_max$ for substrate N-Suc-Ala-Ala-Pro-Phe-pNA were $1.67{\times}10^{-4}\;M$ and 0.6071 $OD_{410}$ per 30 s, respectively. This protease could degrade a broad range of substrates including casein, gelatin, BSA (bovine serum albumin), and nematode cuticle. Moreover, the enzyme could immobilize the free-living nematode Panagrellus redivivus and the pine wood nematode Bursaphelenchus xylophilus, suggesting that it might playa role in infection against nematodes. The encoding gene of Mc1 was composed of one intron and two exons, coding for a polypeptide of 405 amino acid residues. The deduced amino acid sequence of Mcl showed 61.4-91.9% identity to serine proteases from other nematode-trapping fungi. Our results identified that Mcl possessed biochemical properties including optimal reaction condition and substrate preference that are different from previously identified serine proteases.

• 한국표면공학회지
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• 제43권2호
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• pp.105-110
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• 2010

Electroless nickel plating on porous carbon substrate for the application of MCFC electrodes was investigated. Acidic and alkaline bath were used for the electroless nickel plating. The pore sizes of carbon substrates were 16-20 ${\mu}m$ and over 20 ${\mu}m$. The carbon surface was changed from hydrophobic to hydrophilic after immersing the substrate in an ammonia solution for 40 min at $60^{\circ}C$. The contact angle of water was decreased from $85^{\circ}C$ to less than $20^{\circ}$ after ammonia pretreatment. The deposition rate in the alkaline bath was higher than that in the acidic bath. The deposition rate was increased with increasing pH in both acidic and alkaline bath. The content of phosphorous in nickel deposit was decreased with increasing pH in both acidic and alkaline bath. The contents of phosphorous is low in alkaline bath. The minimum concentration of $PdCl_2$ for the electroless nickel plating was 10 ppm in alkaline bath and 5 ppm in acidic bath. The thickness of nickel was not affected by the concentration of $PdCl_2$.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2000년도 제18회 학술발표회 논문개요집
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• pp.60-60
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• 2000

The adhesion interface formation between copper and poly(ethylene terephthalate)(PET), poly(methyl methacrylate)(PMMA) and Polyimide films was treated using Ion assisted reaction system to sequential sputter deposition by High-Frequency ion source. The ion beam modification system used a new type of low power HF ion thruster for space application as new low thruster electric propulsion system. Low power HF ion thruster with diameter 100mm gives the opportunity to obtain beams of Ar+ with currents 20~150 mA (current density 0.5~3.5 mA/cm2) and energy 200~2500eV at HF power level 10~150 W. Using Ar as a working gas it is possible to obtain thrust within 3~8 mN. Contact angles for untreated films were over 95$^{\circ}$ and 80 for Pet, 10o for PMMA and 12o for PI samples as a condition of ion assisted reaction at the ion dose of 10$\times$1016 ions/cm2, the ion beam potential of 1.2 keV and 4 ml/min for environmental gas flow rate. 900o peel tests yielded values of 15 to 35 for PET, 18 to 40 and 12 to 36 g/min. respectively. High resolution X-ray photoelectron spectrocopy is the Cls region for Cu metal on these polymer substrates showed increases in C=O-O groups for polymide, whereas PET and PMMA treated samples showed only C=O groups with increase the ion dose. Finally, unstable polymer surface can be changed from hydrophobic to hydrophilic formation such as C-O and C=O that were confirmed by the XPS analysis, conclusionally, the ion assisted reaction is very effective tools to attach reactive ion species to form functional groups on C-C bond chains of PET, PMMA and PI.

• 접착 및 계면
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• 제1권1호
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• pp.23-29
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• 2000

소수성 고분자 재료 표면의 친수화 처리 및 기능성 작용기 도입은 재료의 응용성을 상당히 확장할 수가 있기 때문에 중요성이 인지되고 있다. 본 연구에서는 기상 UV 그라프트 반응(vapor phase photografting)에 의해 서로 다른 이온성을 가지는 비닐계 단량체들을 PE 표면에 도입하여 표면 친수성을 향상시켰다. 이를 위해 acrylic acid(음이온성), acrylamide(중성), allylalcohol(중성), allylamine(양이온성) N,N-dimethyl aminopropyl acrylamide(양이온성) 등 서로 다른 기능성 작용기를 갖는 친수성 단량체들을 본 실험실에서 고안한 기상 UV 그라프트 반응장치를 이용하여 기체상태에서 PE 표면에 도입하였다. 이들 단량체들이 도입된 PE 표면을 물 접촉각 측정과 ATR-FTIR을 이용하여 분석한 결과, 각 기능성 단량체들이 효과적으로 PE 표면에 그라프트되어 표면의 친수성이 향상되었음을 확인할 수 있었다. 본 연구에 사용한 기상 UV 그라프트 반응은 고분자 표면에 다양한 작용기를 도입하여 표면을 개질할 수 있는 효과적인 방법으로 생각되어진다.

• Korean Chemical Engineering Research
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• 제59권4호
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• pp.639-643
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• 2021

본 연구에서는 3차원 다층 미세유체 디바이스를 제작하기 위한 접착제로서 불소화 에틸렌 프로필렌(fluorinated ethylene propylene, FEP) 나노입자를 연구하였다. FEP 분산 용액을 1500 rpm에서 30초 동안 단순 스핀 코팅하여 기판에 3 ㎛ 두께의 균일하게 분포된 FEP 나노 입자 층을 형성하였다. FEP 나노입자는 300 ℃에서 1시간 동안 열처리 후 소수성 박막으로 변형되었으며, FEP 나노입자를 이용하여 제작된 폴리이미드 필름 기반 미세유체 디바이스는 최대 2250 psi의 압력을 견디는 것을 확인하였다. 마지막으로 기존의 포토리소그래피로 제작하기 어려운 16개의 마이크로 반응기로 구성된 3차원 다층 미세유체 디바이스를 FEP가 코팅된 9개의 폴리이미드 필름을 간단한 1단계 정렬로 성공적으로 구현하였다. 개발된 3차원 다층 미세유체 디바이스는 화학 및 생물학의 다양한 응용을 위한 고속대량 스크리닝, 대량 생산, 병렬화 및 대규모 미세유체 통합과 같은 강력한 도구가 될 가능성이 있습니다.

• Journal of Microbiology and Biotechnology
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• 제32권3호
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• pp.287-293
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• 2022

The hydroxylation of methane (CH₄) is crucial to the field of environmental microbiology, owing to the heat capacity of methane, which is much higher than that of carbon dioxide (CO₂). Soluble methane monooxygenase (sMMO), a member of the bacterial multicomponent monooxygenase (BMM) superfamily, is essential for the hydroxylation of specific substrates, including hydroxylase (MMOH), regulatory component (MMOB), and reductase (MMOR). The diiron active site positioned in the MMOH α-subunit is reduced through the interaction of MMOR in the catalytic cycle. The electron transfer pathway, however, is not yet fully understood due to the absence of complex structures with reductases. A type II methanotroph, Methylosinus sporium 5, successfully expressed sMMO and hydroxylase, which were purified for the study of the mechanisms. Studies on the MMOH-MMOB interaction have demonstrated that Tyr76 and Trp78 induce hydrophobic interactions through π-π stacking. Structural analysis and sequencing of the ferredoxin domain in MMOR (MMOR-Fd) suggested that Tyr93 and Tyr95 could be key residues for electron transfer. Mutational studies of these residues have shown that the concentrations of flavin adenine dinucleotide (FAD) and iron ions are changed. The measurements of dissociation constants (K_ds) between hydroxylase and mutated reductases confirmed that the binding affinities were not significantly changed, although the specific enzyme activities were significantly reduced by MMOR-Y93A. This result shows that Tyr93 could be a crucial residue for the electron transfer route at the interface between hydroxylase and reductase.