• 한국자기공명학회논문지
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• 제14권2호
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• pp.88-104
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• 2010

Human melanocortin-4 receptor (hMC4R) has a critical role in part of energy homeostasis, and their heterozygous mutations related in genetic cause of severe human obesity. In order to study the structure and function of these membrane proteins, it is important to prepare the samples. However, the preparation of transmembrane peptide is seriously difficult and time-consuming. Overexpression and purification of membrane proteins was reported to be difficult due to their innate insoluble and toxic properties. Among the many difficulties, the most important is the difficulty in obtaining sufficient quantities of purified protein. Recently, we succeed to produce large amounts of the second transmembrane domain from the wild-type hMC4R (wt-TM2) and D90N mutant hMC4R (m-TM2) and proposed the structural difference of them in membrane-like environments. In this paper, we demonstrate the optimization procedures to express and purify wt-TM2 or m-TM2 peptides, and solution NMR studies in different detergents to get high-resolution spectra were also described.

• 한국토양비료학회지
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• 제20권4호
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• pp.351-357
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• 1987

상품화(商品化) 되어있는 유기질비료(有機質肥料) Bio-com$^{(R)}$을 토양(土壤)에 시용시(施用時) 토양(土壤)의 pH, Eh, gas 생성(生成), Ammonia 화성율(化成率) 질산화성율(窒酸化成率) 및 미생물(微生物) 변화(變化)에 미치는 영향(影響)을 알기 위하여 일반농가퇴비(一般農家堆肥) 및 양송이 폐상퇴비등(等)을 비교(比較)하여 담수조건(湛水條件)에서 실내시험(室內試驗)한 결과(結果)를 요약(要約)하면 다음과 같다. 1. 삼요소구(三要素區)에 비(比)하여 각종(各種) 퇴비시용(堆肥施用)은 토양(土壤) pH를 상승시키는 경향(傾向)이 있었으며 Bio-com$^{(R)}$ 시용(施用)은 타공시유기물(他供試有機物)과 상이(相異)하지 않했다. 2. 시험후(試驗後) 토양(土壤)의 미생물수(微生物數) 변화(變化)는 삼요소(三要素) 혹은 각종(各種) 퇴비(堆肥)를 삼요소(三要素)와 병용(倂用)하므로서 현저(顯著)한 증가(增加)를 보였으며 농가퇴비시용구(農家堆肥施用區)에서 세균수(細菌數)의 증가(增加)가 현저(顯著)하였다. 3. Bio-com$^{(R)}$ 시용(施用)은 항온기간중(恒溫期間中) 질산화성율(窒酸化成率)을 증가(增加)시켰으며 그율(率)은 타처리(他處理)에 비(比)하여 수배정도(數培程度)높아서 밭작물재배지(作物栽培地)에 시용(施用)이 합리적(合理的)이었다. 4. Bio-com$^{(R)}$ 시용(施用)에 의한 gas의 생성(生成)은 일반퇴비(一般堆肥)나 양송이 폐상퇴비(堆肥) 시용(施用)과 비슷한 경향(傾向)을 보였고 $N_2O$의 생성(生成)은 삼요소시용(三要素施用)보다 낮은양(量)을 보였다.

• Journal of Microbiology and Biotechnology
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• 제19권5호
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• pp.439-446
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• 2009

The biosynthesis study of antibiotics saframycin (SFM) in Streptomyces lavendulae and safracin (SAC) in Pseudomonas fluorescens demonstrated that 3-hydroxy-S-methyl-O-methyltyrosine (3hSmOmTyr), a nonproteinogenic amino acid, is the precursor of the tetrahydroisoquinoline molecular core. In the biosynthetic gene cluster of SAC/SFM, sacD/sfmD encodes a protein with high homology to each other but no sequence similarity to other known enzymes; sacF/sfmM2 and sacG/sfmM3 encode methyltransferases for C-methylation and O-methylation; and sacE/sfinF encodes a small protein with significant sequence similarity to the MbtH-like proteins, which are frequently found in the biosynthetic pathways of non ribosomal peptide antibiotics and siderophores. To address their function, the biosynthetic cassette of 3h5mOmTyr was heterologously expressed in S. coelicolor and P. putida, and an in-frame deletion and complementation in trans were carried out. The results revealed that (i) SfmD catalyzes the hydroxylation of aromatic rings; (ii) sacD/sacF/sacG in the SAC gene cluster and sfmD/sfmM2/sfmM3 in the SFM cluster are sufficient for the biosynthesis of 3h5mOmTyr; and (iii) the mbtH-like gene is not required for the biosynthesis of the 3h5mOmTyr precursor.

• KSBB Journal
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• 제20권6호
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• pp.458-463
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• 2005

Polyurethane foam이 충진된 trickle bed reactor에서 통성혐기성 미생물인 Citrobacter amalonaticus Y19을 이용하여 일산화탄소와 물로부터 연속적인 수소생산을 살펴보았다. C. amalonaticus Y19은 설탕을 탄소원으로 할 때 호기적 조건에서 13 g/L까지 성장하였고 혐기조건에서 CO 가스를 주입하였을 때 약 60시간만에 최대 수소 생산 활성을 나타내었다. TBR 반응기에서 유입가스의 CO의 분압이 증가할수록 혹은 기체 체류시간이 감소할수록 수소 생성속도가 증가하였으나 CO의 전환율은 반대로 감소하였다. 그러나 액상의 유속변화는 반응기 운전 결과에 큰 영향을 주지 못했다. 본 실험에서 얻은 최대 수소 생성속도는 기체 체류시간 25분, 유입 CO 압력 0.4 atm에서 16 mmol/L/hr(전환율 33%)이었다. 이 값은 비슷한 반응기에 대해 보고된 Cowger의 결과보다 약 2배 이상 높은 값으로 통성혐기성균주의 고농도 배양과 다공성 충진물의 사용에 의한 높은 기-액 물질 전달 속도가 그 원인으로 추정되었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제33권2호
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• pp.138-143
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• 2016

In the present study, silk fibroin (SF) was dissolved in $CaCl_2/H_2O/EtOH$ solution at $85^{\circ}C$. After the dissolution, the SF solution was cooled down and stored at $4^{\circ}C$ for 28 d. The stability of the solution's viscosity and electrospinnability was observed to examine the stability of SF molecules during storage in $CaCl_2/H_2O/EtOH$ solution. The viscosities of $SF/CaCl_2/H_2O/EtOH$ solution and SF formic acid solution did not change during 28 days' storage of SF in $CaCl_2/H_2O/EtOH$ solution. The electrospinnability of the SF solution, mean diameter of the electrospun SF fiber, and crystallinity index of electrospun SF web did not change, regardless of the length of the storage period. These results imply that SF molecules do not degrade during 28 days' storage in $CaCl_2/H_2O/EtOH$ solution.

• Animal cells and systems
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• 제13권4호
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• pp.405-409
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• 2009

When DNA double-strand breaks (DSBs) were induced in mammalian cells, many DNA damage response proteins are accumulated at damage sites to form nuclear foci called IR-induced foci. Although the formation of foci has been shown to promote repair efficiency, the structural organization of chromatin in foci remains obscure. BAF53 is an actin-related protein which is required for maintenance of chromosome territory. In this study, we show that the formation of IR-induced foci by $\gamma$-H2AX and 53BP1 were reduced when BAF53 is depleted, while DSB- activated ATM pathway and the phosphorylation of H2AX remains intact after DNA damage in BAF53 knockdown cells. We also found that DSB repair efficiency was largely compromised in BAF53 knockdown cells. These results indicate that BAF53 is critical for formation of foci by $\gamma$-H2AX decorated chromatin at damage sites and the structural organization of chromatin in foci is an important factor to achieve the maximum efficiency of DNA repair.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권1호
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• pp.17-22
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• 2004

A bacterial strain WJ-98 found to produce active extracellular keratinase was isolated from the soil of a poultry factory. It was identified as Paracoccus sp. based on its 16S rRNA sequence analysis, morphological and physiological characteristics. The optimal culture conditions for the production of keratinase by Paracoccus sp. WJ-98 were investigated. The optimal medium composition for keratinase production was determined to be 1.0% keratin, 0.05% urea and NaCl, 0.03% K$_2$HPO$_4$, 0.04% KH$_2$PO$_4$, and 0.01% MgCl$_2$$.$6H$_2$O. Optimal initial pH and temperature for the production of keratinase were 7.5 and 37$^{\circ}C$, respectively. The maximum keratinase production of 90 U/mL was reached after 84 h of cultivation under the optimal culturing conditions. The keratinase from Paracoccus sp. WJ-98 was partially purified from a culture broth by using ammonium sulfate precipitation, ion-exchange chromatography on DEAE-cellulose, followed by gel filtration chromatography on Sephadex G-75. Optimum pH and temperature for the enzyme reaction were pH 6.8 and 50$^{\circ}C$, respectively and the enzymes were stable in the pH range from 6.0 to 8.0 and below 50$^{\circ}C$. The enzyme activity was significantly inhibited by EDTA, Zn$\^$2+/ and Hg$\^$2+/. Inquiry into the characteristics of keratinase production from these bacteria may yield useful agricultural feed processing applications.

• Geosystem Engineering
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• 제21권5호
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• pp.291-296
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• 2018

The aim of this work is to study the effect of aggregate type on the physico-mechanical properties and microstructure of bio-mortar (BM). Three different aggregates such as sand, dolomite and basalt were used. BM was prepared by mixing aggregates with bacterial cells (Sporosarcina Pasteurii) and one equimolar (1 M) of $urea/CaCl_2.2H_2O$. The results proved that the chemical composition and physical properties of aggregates play an important role in the microbial precipitation rate as well as size, morphology and crystallinity of the precipitated calcite, which strongly reflects on the properties of the prepared BM. The BM containing dolomite gave the highest compressive strength and lowest water absorption.

• Bulletin of the Korean Chemical Society
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• 제32권2호
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• pp.455-458
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• 2011

In previous report, with the aid of receptor-oriented pharmacophore-based in silico screening, we characterized three novel hPNMT inhibitors (YPN010, YPN016, and YPN017) and proposed that the hydrogen bonding interaction between inhibitors and side chain of Lys57 is very important to inhibitory activity of hPNMT. To confirm the importance of Lys57, mutant with substitution of Lys57 with Ala was cloned and binding study was performed for a K57A mutant of hPNMT using STD-NMR and fluorescence experiments. The binding constants for three novel inhibitors with mutant hPNMT were dramatically decreased compared to those with wild-type protein. K57A mutant-induced conversion of noradrenaline to adrenaline was suppressed about 95 % compared to wild-type hPNMT. Mutagenic analysis using a K57A mutant confirmed the importance of the Lys57 residue in binding of the inhibitor candidate to hPNMT as well as enzymatic activity of hPNMT, implying that these results are consistent with our binding model.

• Journal of Microbiology and Biotechnology
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• 제28권5호
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• pp.732-738
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• 2018

Novel carbon-based solid acid catalysts were synthesized through a sustainable route from lipid-extracted microalgal residue of Dunaliella tertiolecta, for biodiesel production. Two carbon-based solid acid catalysts were prepared by surface modification of bio-char with sulfuric acid ($H_2SO_4$) and sulfuryl chloride ($SO_2Cl_2$), respectively. The treated catalysts were characterized and their catalytic activities were evaluated by esterification of oleic acid. The esterification catalytic activity of the $SO_2Cl_2$-treated bio-char was higher ($11.5mmol\;Prod.{\cdot}h^{-1}{\cdot}gCat.\;^{-1}$) than that of commercial catalyst silica-supported Nafion SAC-13 ($2.3mmol\;Prod.{\cdot}h^{-1}{\cdot}gCat.^{-1}$) and $H_2SO_4$-treated bio-char ($5.7mmol\;Prod.{\cdot}h^{-1}{\cdot}gCat.^{-1}$). Reusability of the catalysts was examined. The catalytic activity of the $SO_2Cl_2$-modified catalyst was sustained from the second run after the initial activity dropped after the first run and kept the same activity until the fifth run. It was higher than that of first-used Nafion. These experimental results demonstrate that catalysts from lipid-extracted algae have great potential for the economic and environment-friendly production of biodiesel.