• 미생물학회지
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• 제12권4호
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• pp.159-179
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• 1974

In order to investigate morphological and cultural characteristics of strains in species, Flammulina velutipes, the author collected isolates of Flammultina velutipes at 49 locations in Korea and cultivated these isolates on the various kinds of solid media. After investigating the cultural characteristics, appeared on the various media, he obtained the following results : 1. The variation of colors in the fruit bodies is connected with the variation of climatic environments(composite effects between mean temperature in January and number of rain days of 1mm and over precipitation). The author, therfore, can find out the trend that brown type is distributed in the midland climatic region and yellow type in the southern climatic regoin. 2. Two types can be classified into several strains respectively : the strain of abundant or insufficient productivity, and strain of selectivity or non-selectivity of media. 3. According to the results of mutual comparison of soluble mycelial proteins by disc electrophoresis using polyacrylamide gels, each type has special common protein fractions(brown type : band located at 26..5mm position from surface of gel, yellow type : band located at 24.5mm position from surface of gel), and each strain has special protein fractions too. Therefore this phenomenon seems to support the results obtained by the above-stated morphological and cultural studies. 4. In the adaptability of strains to the temperature, every strain has the nature of growing in lower temperature(the optimum temperature of 20.deg.C to 25.deg.C) except that YI-1 strain has the optimum temperatue of $25^{\circ}C$-26^{\circ}C. And mycelial growth of every strain is discontinued at $35^{\circ}C.$ 5. In the adaptability of strains to the H-ion concentration, every strain has wide adaptable range of H-ion concentration, and has optimum range of pH 5.5 to 6.6 in mycelial growth excepting YA01, BI-2 and YI-1. 6. In the utility of carbon sources, the mycelial growth of every strain is very poor on the media containing xylose(average diameter of mycelial growth : 18mm), and most strains utilize favorably sucrose(39mm), maltose(37mm) and dextrose(35mm) in mycelial growth. In the utility of nitrogen sources, every strain utilizes favorably organic nitrogens(36 mm)more than inorganic nitrogens (25 mm), and utilizes fully peptone nad asparagine in organic nitrogens. Especially BA-1, BIK-2 and YA-1 strains grow vigorously on each media containing various carbon and nitrogen sources. 7. The characteristic tests of the number of days required for mycelial growth, the number of days requried for sprout of young bodies, the length of stipe and the number of fruit bodies formation seem to be useful methods in the early selection of the strain of the abundant productivity.

• Applied Biological Chemistry
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• 제35권1호
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• pp.23-29
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• 1992

한국산 파인애플로부터 bromelain을 추출하여 황산암모늄염석, DEAE-cellulose ion-exchange chromatography, gel filtration을 이용하여 약 21배 정제하였고, 정제효소는 전기영동상 단일밴드를 나타내었으며, 분자량은 약 22,000이었다. 정제된 bromelain은 glycine과 serine이 가장 많이 함유되어 있었으며 최적작용 pH와 온도는 6.0, $60^{\circ}C$였다. $pH\;5{\sim}7,\;50^{\circ}C$이하에서 안정성을 보였으며 $Mn^{2+}$에 의해 활성이 촉진되었고, $Mg^{2+},\;Fe^{2+}$ 등의 금속이온에 의해 급격한 활성저해가 관찰되었다. 정제효소는 p-chloromercuribenzoic acid에 의해 저해되어 SH효소로 추측되었으며, Km과 Vmax값는 각각 $5.747{\times}10^{-4}M,\;131.58\;{\mu}g/min$ 이었다. 돈육 조직에 효소를 처리하였을 때 효소 농도가 증가함에 따라 육단백질이 가수분해되어 유리되는 수용성 질소량과 아미노태 질소량이 증가되어 연육효과가 있음이 확인되었다.

• 생명과학회지
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• 제11권5호
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• pp.483-488
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• 2001

최근 다양한 분야에서 생물학적 제제개발에 관한 연구가 활발한다. 본 연구에서는 생물학적 제제개발을 위한 일련의 연구로 Pseudomonas aeruginosa KLP-2가 생산하는 항균성 물질을 동정하고, 생리화학적 특성을 확인하였다. 본 항균성 물질을 전제하여 FAB-MS로 확인한 결과 pyocyanine으로 동정되었으며, 진청색의 needles 성상을 나타내었고 chloroform, methanol, ethyl acetate와 같은 용매에 높은 용해도를 나타내었다. Pyocyanine의 UV spectrum은 methanol에서 최대 흡광치(318 nm)를 보였고, FAB-MS로 확인한 결과 m/z 211에서 protonated molecular ion(M+H)$^{+}$ 이 관찰되었으며, 분자식은 $C_{13}$H$_{10}$ N$_{2}$O로 확인되었다. Pyocyanine 은 Bacillus cereus, Bacillus subtilis, Micrococcus luteus, Rodococcus equi, Staphylococcus aureus, Streptococcus faecalis, E. coli. Kegionella peneumophila, Shigella flexneri, Shigela boydii, Shigella sonnei, NAG Vibro cholerae, Vibrio parahaemolyticus, Vibro vulnificus, Yersinia enterocolitica 및 Saccharomyces cerevisiae 등의 다양한 세균에 대하여 항균활성을 보여 비교적 넓은 항균범위를 나타내었으며, 특히 Legionella pneumophila에 대하여 0.5$\mu$g의 pyocyanine을 함유한 disk가 48mm의 억제환을 타나내어 가장 높은 항균활성을 보였다.

• 한국식품영양학회지
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• 제17권1호
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• pp.47-52
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• 2004

Mammaglobin은 uteroglobin 유전자와 상동성을 가지는 분비 단백질로 인체 유방암 조직에서 과발현된다. 이 단백질은 유방암의 진단, 전이 정도의 진단, 또는 수술 및 항암치료 후 재발 정도의 검색을 위한 하나의 표식자로 가능성을 갖는다. 본 연구는 mammaglobin 유전자를 클로닝하여, 대장균으로부터 발현하고, 발현된 mammaglobin 단백질을 분리하고, 분리된 단백질을 이용하여 항체를 생산하고, 분리된 항체가 mammaglobin에 대한 특이 반응을 갖는지를 확인하였다. 유방암 환자의 조직을 얻은 후 이 조직에서 RNA를 분리한다. 이 RNA로부터 RT-PCR법으로 mammaglobin 유전자를 클로닝하였다. 증폭된 유전자를 NcoI 과 XhoI으로 절단한 후 벡터에 끼워 넣은 후 대장균에 형질 전환시키고 DNA 염기 서열을 결정하였고, 기존의 mammaglobin 유전자의 염기서열과 비교한 결과 동일한 유전자임을 확인하였다. Mammaglobin의 세포 내 발현, 신호 펩타이드를 이용한 분비발현을 위해 pET30, pET20, pET32 벡터를 각각 이용하였다. 3개의 발현시스템으로부터 단백질이 과 발현됨을 확인할 수 있었다. pET30 벡터를 이용하여 성공적으로 발현된 mammaglobin 단백질을 분리할 수 있었다. Ni-NTA affinity chromatography에 이은 DEAE-ion exchange chromatography 분리 방법에 의해 수용성 발현 단백질인 thioredoxin-mammaglobin을 정제할 수 있었고 이 융합 단백질로부터 enterokinase를 이용하여 mammaglobin 단백질만을 분리하였다. 토끼에 분리된 mammaglobin을 complete adjuvant와 혼합하여 면역한 후 두 번 boosting하여 polyconal 항체를 얻었다. Westernblot immuno 분석을 한 결과 생산된 항체가 mammaglobin 단백질과 특이적 항원항체반응을 보임을 관찰하였다. 향후 이 항체를 이용하여 진단용 시약의 개발이나 항암제 개발 등을 위해 연구가 진행될 것이다.

• Korean Chemical Engineering Research
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• 제51권4호
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• pp.443-454
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• 2013

콜로이드 실리카와 가용성 실리카를 이용하여 나트륨이 첨가되지 않은 다양한 금속이온 첨가 MCM-41 촉매를 제조하였다. 전이금속 이온인 $V^{5+}$, $Co^{2+}$ 및 $Ni^{2+}$이 MCM-41에 첨가되었을 경우 기공벽 내의 실리콘 이온과 등방치환을 하여 실리카 기공벽 내에서 독립된 단일 활성점을 형성하여 우수한 환원 및 활성 내구성을 보였다. 수소 승온 환원법을 이용하여 Co-MCM-41 촉매의 기공 곡률 반경효과에 대해 검토해 본 결과, 적절한 환원 처리와 기공 크기 및 pH 조절에 따라 코발트 금속입자의 크기를 1nm 이하의 범위에서 조절할 수 있었으며, 이 미세 금속 입자들은 표면 금속이온들과의 결합으로 인해 상당한 고온 안정성이 있음을 발견하였다. 완전 환원 후에도 비정형 실리카의 부분 덮힘으로 인해 금속 입자들의 표면 이동 및 뭉침 현상이 현저히 저하되는 것을 볼 수 있었다. 이들 촉매의 반응 예로 금속 입자 크기에 민감한 단일층 탄소 나노튜브의 합성을 Co-MCM-41을 이용하여 실시하였고, 금속 입자의 안정성 시험반응으로 Co 및 Ni-MCM-41을 이용한 CO 메탄화 반응, V-MCM-41을 이용한 메탄올 및 메탄의 부분 산화반응 및 기공곡률 반경이 촉매활성에 미치는 영향 등을 살펴보았다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2001년도 제32회 학술심포지움
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• pp.67-86
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• 2001

A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.663-669
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• 2008

The solvent-tolerant bacterium Enterobacter sp. VKGH12 is capable of utilizing n-butanol and contains an $NAD^+$-dependent n-butanol dehydrogenase (BDH). The BDH from n-butanol-grown Enterobacter sp. was purified from a cell-free extract (soluble fraction) to near homogeneity using a 3-step procedure. The BDH was purified 15.37-fold with a recovery of only 10.51, and the molecular mass estimated to be 38 kDa. The apparent Michaelis-Menten constant ($K_m$) for the BDH was found to be 4 mM with respect to n-butanol. The BDH also had a broad range of substrate specificity, including primary alcohols, secondary alcohols, and aromatic alcohols, and exhibited an optimal activity at pH 9.0 and $40^{\circ}C$. Among the metal ions studied, $Mg^{2+}$ and $Mn^{2+}$ had no effect, whereas $Cu^{2+},\;Zn^{2+}$, and $Fe^{2+}$ at 1 mM completely inhibited the BDH activity. The BDH activity was not inhibited by PMSF, suggesting that serine is not involved in the catalytic site. The known metal ion chelator EDTA had no effect on the BDH activity. Thus, in addition to its physiological significance, some features of the enzyme, such as its activity at an alkaline pH and broad range of substrate specificity, including primary and secondary alcohols, are attractive for application to the enzymatic conversion of alcohols.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.730-734
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• 2008

Barley ${\alpha}$-amylase genes, amy1 and amy2, were separately cloned into the expression vector of $pPICZ{\alpha}A$ and recombinant Pichia strains were established by homologous recombination. Both AMYs from Pichia shared almost identical hydrolysis patterns on short maltooligosaccharides to result in glucose, maltose, or maltotriose. Against insoluble blue starch, AMY1 showed the highest activity at 0.1-5 mM calcium concentration, whereas 15-20 mM was optimal for AMY2. On the hydrolysis of soluble starch, unexpectedly, there was no significant difference between AMYs with increase of calcium. However, the relative activity on various starch substrates was significantly different between AMYs, which supports that the isozymes are clearly distinguished from each other on the basis of their unique preferences for substrates.

• KEPCO Journal on Electric Power and Energy
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• 제2권3호
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• pp.437-445
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• 2016

This paper discussed the effect of ammonia concentration adsorbed on fly ash for the ammonia emission as AAFA (Ammonia Adsorbed Fly Ash) produced from coal fired plants due to operation of NOx reduction technologies was landfilled with distilled or sea water at closed and open systems, respectively. Ammonia bisulfate and sulfates adsorbed on fly ash is highly water soluble. The pH of ammonium bisulfate and sulfate solution had significant effect on ammonia odor emission. The effect of temperature on ammonia odor emission from mixture was less than pH, the rate of ammonia emission increased with increased temperature when the pH conditions were kept at constant. Since AAFA increases the pH of solution substantially, $NH_3$ in the ash can release the ammonia order unless it is present at low concentration. $NH_4{^+}$ ion is unstable in fly ash and water mixtures of high pH at open system, which is changed to nitrite or nitrate and then released as ammonia gas. The proper conditions for < 20 ppm of ammonia concentration released from the AAFAs landfilled in ash pond were explored using an open system with sea water. It was therefore proposed that optimal operation to collect AAFA of less than 168 ppm ammonia at the electrostatic precipitator were controlled to ammonia slip with less than 5 ppm at SCR/SNCR installations, and, ammonia odor released from mixture of fly ash of 168 ppm ammonia with sea water under open system has about 20 ppm.

• KSBB Journal
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• 제19권5호
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• pp.368-373
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• 2004

The optimal condition for the production of a glucan and a fructan synthesizing enzymes from Leuconostoc mesenteroides NRRL B-1149 were studied based on the different medium compositions. Response surface methodology was applied to find the optimistic condition showing the relationship between the fermentation response (enzyme activities) and the fermentation variable concentrations of yeast extract, peptone concentration, K2HP04 concentration and sucrose. Optimum medium composition for both enzymes production was $0.75\%$ yeast extract, $0.72\%$ peptone, $1\%$ K2HP04 and $2.17\%$ sucrose. Using this medium, the activities produced in culture was 0.90 U/m~ for glucosyltransferase (GTase) and 0.96 U/ml for fructosyltransferase (FTase). After purification of 1149FTase by consecutive chromatographies using Sephadex G-150 and DEAE-Sepharose, a 1149FTase of 210 kDa on $7\%$ polyacrylamide gel was isolated and it synthesized soluble fructan. The 1149GTase showed a band of 180 kDa on $8\%$ polyacrylamide gel after purification using Bio-Gel P-100 gel chromatography and DEAE-Sepharose ion exchange chromatography and it synthesized insoluble glucan. The linkages of polymers were determined by methylation using Hakomori reagent and following NMR analysis. The glucan was composed of a(1~6) and a(1~3) linkages and the fructan was levan.