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내열성 Chitinase 생산균주의 분리 및 효소생산 특성
홍범식,윤호근,신동훈,조홍연 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5
자연계 고온환경으로부터 내열성 chitinase 우수하고 반응산물로 N-acetyl-D-glucosamine 이량체(GlcNAc)_2를 생산하는 균주를 분리 선별하고 Bacillus licheniformis KFB-C14로 동정하였다. 선별균주의 효소생산 특성은 탄소원으로서 효소기질인 colloidal chitin이 첨가될 때만이 생합성이 유도되었으며 유도제의 첨가시기에 의해 효소생산이 크게 영향을 받았다. 각종 무기, 유기태 질소원 중 yeast extract가 활성과 비활성을 각각 약 2배 증가시켰으며 높은 친화도를 나타내었다. 균의 최대생육과 효소의 최대생산온도는 55℃이었다. 본 균주의 내열성 chitinase 생산에 미치는 최적배양조건은 1.2% colloidal chitin, 0.15% K_2HPO_4, 0.05% KH_2PO_4 0.01% MgSO_4·7H_2O, 0.1% yeast extract, pH 6.5의 배지를 55℃, 150rpm에서 40시간 회전진탕배양 하였을 때로 3.89 units/ml의 효소활성과 7.4 units/mg의 비활성을 나타내었다. A strain capable of producing thermostable chitinase suitable for chitooligosaccharide production was isolated from high temperature environment and identified as Bacillus licheniformis. The chitinase from Bacillus licheniformis KFB-C14 was only induced by addition of colloidal chitin into the basal medium as carbon source, showing the decrease of the chitinase production by supplemental addition of other carbon sources into the medium containing 1.0% colloidal chitin. Among organic and inorganic nitrogen sources, yeast extract was the most effective for the increase of total activity and specific activity, and had high affinity for the enzyme production. The optimum temperature of cell growth and thermostable chitinase production was 55℃. The optimum culture medium was composed of 1.2% colloidal chitin, 0.15% K_2HPO_4, 0.05% KH_2PO_4 0.01% MgSO_4·7H_2O, 0.1% yeast extract (pH 6.5). Bacillus licheniformis KFB-C14 produced the thermostable chitinase of 3.89 units per culture fluid and 7.4 units per mg protein under rotary shaking at 150 rpm for 40 hr.
Bacillus licheniformis KFB-C14가 생산하는 내열성 Chitinase의 정체 및 특성
홍범식,윤호근,신동훈,조홍연 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5
Bacillus licheniformis KFB-C14가 생산하는 내열성 chitinase를 30~70% ammonium sulfate fractionation, DEAE-Toyopearl 650M, Butyl-Toyopearl 650M, TSK-Gel Toyopearl HW-55F에 의해 정제도 66배, 수율 21%로 전기영동적으로 균일하게 정제하였다. 정제 단백질은 gel permeation chromatography에 의해 86,000±2,000의 분자량을 나타내었으며, SDS 전기영동에 의해 밝혀진 본 효소의 subunit 구조는 monomer였다. 효소 단백질의 안정성을 검토한 결과 80℃에서 30분 열처리에 의해 56%, 37℃에서 20분간 40% ethanol과 ethyl acetate, 단백질 변성제 등의 처리시에도 50% 이상의 잔존활성을 나타냄으로써 공업적으로 유용성이 높은 안정한 단백질로 판명되었다. 효소반응의 최적 pH와 온도는 pH 6.0과 60℃이었고 Mn^2+ 이온에 의해 효소 활성이 저해되었으나 EDTA, N-ethylmaleimide, p-chloromercuribenzoate 등에 의한 활성감소는 관찰되지 않음으로써 금속효소 또는 thiol계 효소에 속하지 않음을 알 수 있었다. 본 효소는 colloidal chitin, 시판용 chitin에는 반응성이 높았으나 exo형 chitinase의 대표적인 기질인 p-nitrophenyl-2-aectamido-2deoxy-β-glucopyranoside, NN'-diacetylchitobiose에는 전혀 반응성을 보이지 않는 전형적인 endo형의 chitinase였다. 본 효소는 colloidal chitin으로부터 주로 (GlcNAc)_2를, 반응시간 경과에 따라 (GlcNAc)_1과 (GlcNAc)_3을 생성하는 반응성을 보였다. Chitinase (EC 3.2.1.14) from culture fluid of Bacillus licheniformis KFB-C14 was purified 66-folds to homogenity in overall yield of 21% by ammonium sulfate fractionation, DEAE-Toyopearl, Butyl-Toyopearl and TSK-Gel HW-55F column chromatography. The enzyme protein had a molecular weight of about 86,000 and was composed of one subunit. The enzyme was significantly stable not only at high temperature but also on treatment with organic solvents and protein denaturants such as SDS, urea and guanidine·HCl. The optimum temperature and pH for reaction was 60℃ and 6.0,respectively. The enzyme activity was inhibited by only Mn^2+ ion, but not inhibited by EDTA, N-ethylmaleimide and pCMB. The enzyme had high activity with colloidal chitin (V_max: 421) and commercial chitin (V_max :480), but not with typical substrates of exo type chitinase. The thermostable chitinase had an useful reactivity for producing functional chitooligosaccharide, showing the production of (GlcNAc)_1, (GlcNAc)_3, and (GlcNAc)_2 as major product.
한의진단명과 진단요건의 표준화 연구 III : 3차년도 연구결과 보고
최선미,양기상,최승훈,박경모,박종현,심범상,김성우,노석선,이인선,정진홍,이진용,김달래,임형호,김윤범,박성식,송태원,김종우,이승기,최윤정,신순식 한국한의학연구원 1997 한국한의학연구원논문집 Vol.3 No.1
The diagnostic requirements were suggested and explained regarding the systems of differentiation of symptoms and signs in the third year study of standardization and unification of the terms and conditions used for diagnosis in oriental medicine. The systems were as follows : - analyzing and differentiating of epidemic febrile disease - analyzing and differentiating in accordance with the Sasang constitution medicine based on four-type recognition - differentiation of disease according to pathological changes of Chong and Ren channels - standards for diagnosis of women's disease - standards for diagnosis of children's disease - standards for diagnosis of motor and sensor disturbance (-muscle. born, joint, etc.) - standards for diagnosis of neuropsychiatric disease - standards for diagnosis of five sense organ disease - standards for diagnosis of external disease The indivisual diagnosis pattern was arranged by the diagnostic requirements in the following order : another name, notion of diagnosis pattern, index of differentiation of symptoms and signs, the main point of diagnosis, analysis of diagnosis pattern, discrimination of diagnosis pattern, prognosis, a way of curing a disease, prescription, herbs in common use, disease appearing the diagnosis pattern, documents. The standards for diagnosis of each disease was arranged by the diagnostic requirements in the following order : another name, notion of disease, the main point of diagnosis, analyzing and differentiating of disease, analysis of disease, discrimination of disease, prognosis, a way of curing and prescription of disease, disease in western medicine appearing the disease in oriental medicine, documents.
Hong, Giyoung,Hong, Soon Man,Koo, Chong Min,Baek, Bum Ki,Lee, Hong-shik,Lee, Youn-Woo American Chemical Society 2015 INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH - Vol.54 No.48
<P>De-crosslinking of silane-cross-linked polyethylene (S-XLPE) in supercritical methanol was investigated. Changes in gel content and molecular weight with reaction temperature and reaction time were analyzed in a batch reactor, and kinetic parameters were calculated from the experimental data. The reaction time for de-crosslinking shortened as the reaction temperature increased and the complete de-crosslinking of S-XLPE was achieved at 320 °C and 20 min. The de-crosslinking rate of S-XLPE fit well with the first order reaction model. In addition, the molecular weight of de-cross-linked polyethylene (DXPE) decreased further as reaction temperature and reaction time increased when the reaction temperature exceeded 360 °C relative to that of the original polyethylene before being cross-linked. The decreasing molecular weight trend also fit well with the first order reaction model.</P>
Lee, Hong-shik,Jeong, Ju Hyeong,Hong, Giyoung,Cho, Hang-Kyu,Baek, Bum Ki,Koo, Chong Min,Hong, Soon Man,Kim, Jaehoon,Lee, Youn-Woo American Chemical Society 2013 INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH - Vol.52 No.20
<P>The effect of various solvents on the de-cross-linking reaction of cross-linked polyethylene under subcritical and supercritical conditions was studied. The gel content of the de-cross-linked polyethylene decreased from 60 to 0.8–2.5% at 380 °C within 5 min in all of the solvents tested in this study. When water, ethanol, and 2-propanol were used, the <I>M</I><SUB>w</SUB> of the de-cross-linked polyethylenes decreased from 349,000 to 200,000–227,000 g/mol, and much lower <I>M</I><SUB>w</SUB> values of 70,000–90,000 g/mol were observed when acetone and diisopropyl ether were used. The de-cross-linked polyethylenes treated in the different solvents had a very similar chemical structure to raw polyethylene. According to a kinetic study using a first-order reaction model, the de-cross-linking rate in each solvent increased in the order 2-propanol < ethanol < <I>n</I>-hexane < water < methanol ≪ diisopropyl ether ≪ acetone at 350 °C.</P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ie4006194'>ACS Electronic Supporting Info</A></P>
자궁내막의 이상 유무 평가 시 질식 자궁초음파와 초음파 자궁조영술의 임상적 유용성
홍재식 ( Jae Shik Hong ),송명섭 ( Myung Seop Song ),임재윤 ( Jae Youn Lim ),이현정 ( Hyun Jung Lee ),안가영 ( Ka Young Ahn ),홍준식 ( Jun Shik Hong ),성석주,김주명 ( Joo Myung Kim ),윤재범 ( Jae Bum Yeon ),주관영 ( Kwan Young Joo 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.3
Hong, Mee-Suk,Kim, Hye-Kyung,Shin, Dong-Hoon,Song, Dae-Kyu,Ban, Ju Yeon,Kim, Bum Shik,Chung, Joo-Ho The Korean Society of Toxicogenomics and Toxicopro 2008 Molecular & cellular toxicology Vol.4 No.4
Obesity is an increasing worldwide health problem that is strongly related to the imbalance of food intake and energy metabolism. It was well-known that several substances in the hypothalamus regulate food intake and energy metabolism. We planned an integrative study to elucidate the mechanism of the development of obesity. Firstly, to find candidate genes with the marvelous effect, the different expression in the hypothalamus between ob/ob and 48-h fasting mice was investigated by using DNA microarray technology. As a result, we found 3 genes [peroxisome proliferator activated receptor, gamma, coactivator 1 alpha (Ppargc1a), calmodulin 1 (Calm1), and complexin 2 (Cplx2)] showing the different hypothalamic expression between ob/ob and 48-h fasting mice. Secondly, a genetic approach on PPARGC1A gene was performed, because PPARGC1A acts as a transcriptional coactivator and a metabolic regulator. Two hundred forty three obese female patients with body mass index (BMI)${\geq}$25 and 285 control female subjects with BMI 18 to<23 were recruited according to the Classification of Korean Society for the Study of Obesity. Among the coding single nucleotide polymorphisms (cSNPs) of PPARGC1A, 2 missense SNPs (rs8192678, Gly482Ser; rs3736265, Thr612Met) and 1 synonymous SNP (rs3755863, Thr528Thr) were selected, and analyzed by PCR-RFLP and pyrosequencing. For the analysis of genetic data, chi-square ($X^2$) test and EH program were used. The rs8192678 was significantly associated with obese women (P<0.0006; odds ratio, 1.5327; 95% confidence interval, 1.2006-1.9568). Haplotypes also showed significant association with obese women ($X^2$=33.28, P<0.0008). These results suggest that PPARGC1A might be related to the development of obesity.
Alkaline Protease 를 생산하는 Bacillus subtilis 의 원형질체 융합
홍범식,양한철,성하진,최양미,이태경 한국농화학회 1989 Applied Biological Chemistry (Appl Biol Chem) Vol.32 No.4
To improve alkaline protease producing strain by protoplast fusion, a strain of Bacillus sp. was treated with 100㎍/㎖ NTG (N-methyl-N'-vitro-N-nitrosoguanidine) for 45 minutes and mutants of Bacillus subtilts Arg^-, Try^-, His^- and Ade^- were isolated. The frequency of protoplast formation was about 99%, when teas of exponential phase were treated with 200㎍/㎖ lysozyme at 42℃ for 10∼30 minutes. In a regeneration medium containing 0.3M sodium succinate, 2.0% polyvinylpyrrolidone, 0.5% casamino acid, 10mM MgCl₂ and 20mM CaCl₂, regeneration frequency cf the isolated Bacillus subtilis strains was 25.2%. The fusion frequency between mutant, was from 2.1×10^(-5) to 8.1×10^(-5) under optimum condition.