• 한국토양비료학회지
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• 제34권5호
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• pp.333-341
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• 2001

퇴비화 촉진 미생물인 Bacillus subtilis LYH201균주가 분비하는 섬유소 분해효소를 분자생물학적으로 연구한 결과는 다음과 같다. 섬유소를 분해하는 유전자는 유전자은행에 의해 구한 약 5,000개의 clone 중 CMC 배지 상에서 활성을 가지는 clone을 선발하여 bglC(pLYH7-39)로 명명하였다. 섬유소를 분해하는 bglC 유전자는 Pvu II, EcoRI, SspI의 제한효소 site를 가지고 있었으며, BglC는 Clostridium acetobutylicum GUN_CLOAB(P15704)와 57%의 identity와 71%의 homology를 나타내어 상동성이 가장 높았으며, CMC-SDS-PAGE 분석으로 56 kDa의 분자량을 나타냈고, 온도는 $50^{\circ}C$, pH는 7에서 활성이 가장 높았다.

• 생태와환경
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• 제46권2호
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• pp.276-288
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• 2013

본 연구는 질소 시비에 의해 증가된 토양 질소가 식물의 성장 및 식물체의 화학적 조성에 미치는 영향과 이로 인한 분해에서의 변화를 확인하고자 야외성장실험과 분해실험을 진행하였다. 온실에서 질소 시비구와 비시구 토양에 각각 벼를 재배하였으며 식물이 성숙한 뒤 수확하여 C, N, lignin, cellulose 함량을 측정하였다. 대조구와 질소 처리구 토양에서 재배된 식물의 개체 당 평균 건중량은 각각 0.70 g, 1.32 g로 질소 시비에 의해 1.9배 증가하였다. 식물체의 N 및 C 함량은 질소 시비에 의해 증가하였고 lignin, C/N, lignin/N, cellulose/N은 감소하였다. 이후, 수확된 식물의 지상부는 microcosm 분해실험에 이용되었으며, 분해 식물체에서 건중량의 변화, microbial biomass C와 microbial biomass N, 그리고 dehydrogenase와 urease 활성을 측정하고, 분해과정 중 발생하는 $CO_2$의 양을 정량하였다. 대조구 토양에서 분해시킨 대조구 식물체와 질소 처리구 식물체, 그리고 질소를 처리한 토양에서 분해시킨 질소 처리구 식물체의 잔존량은 각각 초기 건중량의 53.0%, 47.1%, 53.6%를 나타내었다. 질소 시비는 식물체에서 N 함량을 높이고 C/N 및 lignin/N을 낮추어 식물체의 분해를 촉진하였으나, 분해 과정에서의 토양 질소처리는 분해를 억제하였다. 질소 시비에 의해 토양에서 microbial biomass C와 dehydrogenase 활성은 감소하였고, 반면에 microbial biomass N과 urease 활성은 증가하였다. 분해 중 발생한 $CO_2$의 양은 30일 이후부터 질소 시비에 의해 감소하였다. 분해 식물체에서 측정된 microbial biomass C는 질소 처리에 의해 초기에 증가하였으나 이후 저해되는 양상을 나타냈으며 microbial biomass N은 유의한 차이를 보이지 않았다. 질소 시비에 의해 분해 식물체에서 dehydrogenase 활성은 저해되었으며 urease는 분해 초기에 가장 높은 활성을 보였으나 분해 후기에 현저한 감소를 나타냈다. 본 실험에서 질소 시비는 식물의 성장을 증가시키고 식물체의 N 함량을 높여 화학적 조성의 변화를 일으키며 분해율을 증가시키나 분해 단계에서 질소의 시비는 미생물의 활성을 억제시켜 분해를 저해하는 결과를 나타내었다.

• 미생물학회지
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• 제55권2호
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• pp.177-179
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• 2019

미국 뉴저지주 중위도 산림토양에서 부식산(천연 복합유기화합물인 부식질의 주요 구성성분) 분해능이 있는 세균 균주 Pseudomonas kribbensis CHA-19를 분리하였으며, 이후 또 다른 토양 유기물인 리그닌과 리그닌 유래의 페룰산(ferulic acid)과 바릴린산(vanillic acid)의 분해능을 확인하였다. 부식질 초기 저분자화 효소(예, dye-decolorizing peroxidase와 laccase-like multicopper oxidase)와 부식질 유래의 다양한 저분자 분해산물들을 분해하는 효소(예, vanillate O-demethylase와 biphenyl 2,3-dioxygenase)를 탐색하기 위해 CHA-19 게놈염기서열을 분석하였다. 최종 확보한 효소유전자 정보는 토양세균의 부식질 분해경로 제안에 사용되었다.

• 한국의류학회지
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• 제17권3호
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• pp.491-505
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• 1993

The influences of protease on the removal of various protein soils from cotton fabrics were studied. The human epidermal stratum corneum, hemoglobin and casein were used as protein soils. The soiled fabrics were denatured by steaming for 30 min. before washing and laundered using Terg-O-Tometer under washing conditions. The removal efficiency was evaluated by analysis of protein on the fabrics before and after washing by means of copper-Folin method. The relations between the removal and the characteristics of protease were discussed. Also the degradation of protein were examined by microscopy. The seperation of human epidermal stratum corneum after hydrolysis was examined by SDS-PAGE. The results obtained were as follow : 1. The protein from the soiled cotton fabric was removed effectively by adding protease. The removal of protein was increased in proportion to increasing of the enzyme concentration up to a certain point, but it began to decrease above the point. The removal effect was high in the order of casein>human epidermal stratum corneum>hemoglobin. Especially the protein was more effectively removed in ADS solution(pH 9.5) containing enzyme. 2. When protease was used with ADS. the removal of protein was efficiently showed in relatively short time(5~15min.) compared to using ADS only. It is due to the properties of this enzyme that reacts with very short time. 3. Even at low temperature the removal efficiency of enzyme was relatively higher compared with the activity of enzyme. The removal of protein soil was increased up to a maximum near $50^{\circ}C$, and then decreased. 4. The removal of protein by protease was improved with the increase of alkalinity in the pH range from 9.5 to 11.0 but it began to decrease above pH 11.0. 5. According to the increase of mechanical agitation, the removal effect was increased. But the removal efficiency of protease was more effective compared with the agitation in detergency. 6. According to the SDS-PAGE separation and micrograph it was confirmed that the human epidermal corneum was effectively hydrolysed by the enzyme added. So the fragments of protein were removed more efficiently by means of the interfacial reaction of AOS.

• 한국미생물·생명공학회지
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• 제8권1호
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• pp.27-32
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• 1980

본 실험에서 얻은 성과를 요약하면 다음과 같다. \circled1 Furfuryl alcohol를 단일탄소원으로 이용하여 ADH를 생산하는 세균을 토양중에서 분리하고, 균학적 성질에 따라 Pseudomonas sp.라 동정하였다. \circled2 본 효소는 disc 전기영동적, SDS 전기영동적, 초원침강적, 분석에서 완전히 균일하였다. \circled3 본 효소의 침강계수는 7.6S, 반응최적활성 pH는8.5~9.0에 위치하였으며, 최적활성온도는 45$^{\circ}C$ 이었다. \circled4 4개의 동일 subunit (M.W 30,000)가 회합하여 본 활성효소를 형성하는 tetramer이며, 분자량은 개략 120,000이다. \circled5 본 효소의 subunit 1분자는 266개의 amino 산 residues로 구성되고 있다.

• 한국미생물·생명공학회지
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• 제10권1호
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• pp.45-52
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• 1982

A fungus with the highest protopectinase productivity was selected among 205 strains isolated from the soil and identified as a Verticillium sp. The Verticillium sp. was cultivated on wheat bran and the crude extruct of its culture medium showed the highest protopectinase activity on the following conditions: 3 days of cultivation time, 27$^{\circ}C$ of cultivation temperature, 1.2 $m\ell$/g wheat bran of water content, and reinforcement of ammonium nitrate and calcium chloride at the concentration of 0.5 and 0.02%, respectively. The optimum conditions for pectin production from Citrus peel pulp by the protopectinase were consequently obtained as follows: 20$m\ell$/g of liquid volume-to-pulp weight ratio, 4$0^{\circ}C$ of reaction temperature, and 4 of reaction pH. The higher the enzyme concentration, the better the yield of pectin and the shorter the reaction time. Total 45.6mg of pectin/g peel was produced by 1 hour reaction at the enzyme concentration of 10.5 units/$m\ell$. Molecular weight of the pectin produced by the enzyme was estimated to be about 62,000 by Smit and Bryant's method.

• BMB Reports
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• 제40권3호
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• pp.333-340
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• 2007

Gene encoding cyclodextrin glycosyltransferase (CGTase), from thermotolerant Paenibacillus sp. T16 isolated from hot spring area in northern Thailand, was cloned and expressed in E. coli (JM109). The nucleotide sequences of both wild type and transformed CGTases consisted of 2139 bp open reading frame, 713 deduced amino acids residues with difference of 4 amino acid residues. The recombinant cells required 24 h culture time and a neutral pH for culture medium to produce compatible amount of CGTase compared to 72 h culture time and pH 10 for wild type. The recombinant and wild-type CGTases were purified by starch adsorption and phenyl sepharose column chromatography and characterized in parallel. Both enzymes showed molecular weight of 77 kDa and similar optimum pHs and temperatures with recombinant enzyme showing broader range. There were some significant difference in pH, temperature stability and kinetic parameters. The presence of high starch concentration resulted in higher thermostability in recombinant enzyme than the wild type. The recombinant enzyme was more stable at higher temperature and lower pH, with lower $K_m$ for coupling reaction using cellobiose and cyclodextrins as substrates.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.452-457
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• 2001

The glutaryl 7-aminocephalosporanic acid (glutaryl 7-ACA) acylase was purified from Pseudomonas diminuta KAC-1 cells isolated from soil, and characterized. The acylase was purified by procedures including ammonium sulfate fractionation and column chromatographies on DEAE-Sepharose, Phenyl-Sepharose, Q-Sepharose, and Superose 12H/R. The negative acylase was found to be composed of two subunits with molecular masses of approximately 55 kDa and 17 kDa, respectively. The isoelectric point of the enzyme was 4.0. The specific activities of the purified acylase were 8.0 and 7.0 U/mg on glutaryl 7-ACA and glutaryl 7-aminodesacetoxy cephalosporanic acid (glutaryl 7-ADCA), respectively, and $K_m$ values were 0.45 mM for glutaryl 7-ADCA and 0.67 mM for glutaryl 7-ADCA. The enzyme had a pH optimum at 8.0 and a tmperature optimum at $40^{\circ}C$. The acylase catalyzed the synthesis of glutaryl 7-ACA from glutaric acid and 7-ACA as well as the hydrolysis of glutaryl 7-ADCA, although the reaction rate of the synthesis was slower than that of the hydrolysis. In addition, it was found that the enzyme had a glutaryl transferase activity, thereby transferring the glutaryl group from one cephalosporin nucleus to another.

• 미생물학회지
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• 제22권4호
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• pp.215-222
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• 1984

Rhodotorula gl$\varkappa$tinis 세포벽에 작용하는 용해 효소 생산곰팡이를 토양으로부터 분리하였고, Aspergillus f'||'&'||'micro;mig$\alpha$tus에 속하는 species로 동정되었다. 이 세포벽 용해효소는 세표외 유도효소였으며 lytic polysaccharidase 와 protease로 구성되어 생세포 용해에 공동으로 착용하였다. 이 lytic polysaccharidase는 Ascomycetous 효모에서의 주 구성 결합인 ${\beta}-1,3-$와 ${\beta}-1$, 6-glucan에는 작용치 않았다. 이 효소는 생세포에는 역가가 낮았지만 R. glutinis의 분획된 세포액에는 protease의 도움없이 작용할 수 있었다.

• Preventive Nutrition and Food Science
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• 제1권1호
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• pp.121-126
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• 1996

A bacterial strain LC-413, producing extracellular inulin fructotransferase which converts inulin into di-D-fructofuranose dianhydride(DFAIII) and amount of oilgosaccharides, was isolated from soil and pre-sumed as Flavobacteium sp. LC-413. The enzyme production was induced by inulin as carbon source and enhanced by the addition of 0.3% malt extract and 0.2% {TEX}$NaNO_{3}${/TEX} as nitrogen source. The enzyme activity in the culture supernatant reached at the maximum, 78.6units/ml, after 11 hours of cultivation in the medium composition of 1.5% inulin, 0.2% {TEX}$NaNO_{3}${/TEX}, 0.05% {TEX}$K_{2}${/TEX}{TEX}$HPO_{4}${/TEX}, 0.05% {TEX}$MgSO_{4}${/TEX}.7{TEX}$H_{2}${/TEX}O, 0.05% KCI, a trace amount of {TEX}$FeSO_{4}${/TEX}.7{TEX}$H_{2}${/TEX}O, and 0.3% malt ext. at 3$0^{\circ}C$. The oilgosaccharide produced by enzyme reaction from inulin was identified as DFA III by and {TEX}${13}^C${/TEX}-NMR spectrosocpy.