• 산업식품공학
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• 제21권2호
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• pp.103-109
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• 2017

DEAE sephacel anion chromatography 및 SP sepharose cation chromatography에 의해 Streptomyces violaceoruber 유래 alginate lyase의 정제를 수행하여 비활성 14.59 units/mL 정제배율 40.64배를 나타내었다. Tricine SDS-PAGE에 의한 단일밴드를 확인하였고, 분자량은 23.3 kDa으로 결정되었다. 정제효소에 의해 sodium alginate를 가수분해하여 1차 activated carbon column chromatography와 2차 bio gel P-2 gel filtration에 의해 당가수분해물을 분리 회수하여 TLC와 FACE를 통해 중합도를 확인하고 Timell's method에 의해 hetero type M/G-oligosaccharide 중합도 6, 8로 결정되었다. B. animalis, B. bifidum, B. breve, B. infantis, B. longum와 L. acidophilus, L. casei, L. reuteri에 생육활성에 대한 중합도 6, 8의 영향을 검토하기 위하여 modified-MRS media에 탄소원으로 중합도 6, 8를 대체하여 생육활성을 비교한 결과 B. longum에서는 D.P. 6 M/G-oligosaccharide를 탄소원으로 대체한 경우 표준 MRS배지와 비교하여 4.25배, D.P. 8에서 6.44배의 상대활성을 나타내어 가장 우수한 생육활성을 나타내었으며, B. bifidum의 경우에서도 D.P. 6에서 3.27배, D.P. 8에서 5.4배의 상대활성을 나타내었다. 이외에도 B. animalis, B. breve그리고 L. casei에 있어서도 D.P. 8의 경우 3배의 상대활성을 나타내었으나, L. reuteri에 대한 D.P. 8의 경우에서는 표준 MRS media와 비교하여 0.29배로 감소하였다. 결과적으로 D.P. 8의 올리고당이 D.P. 6의 올리고당보다 생육활성에 크게 기여하는 것으로 나타났다.

• Parasites, Hosts and Diseases
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• 제30권4호
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• pp.289-298
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• 1992

Acanthnmoeba sp. YM-4의 영양형은 사상의 침상위족을 내어 전형적인 각canthamoeba 속의 특징이 관찰되었으며, 포낭(씨스트)은 외벽이 등근혐이었고 내벽이 단순한 구형이거나 다소 불규칙한 2개의 세포벽을 갖고 있었다. SDS-PAGE 결과, Acanthamoeha sp. Yhf-4는 약 16개의 주요 단백질 분획이 관찰되었는데, 2차원 전기영동 결과와 함께 Acanthamoeba sp. YM-4의 단백질 분획은 A. culbertsoni와 거의 같은 양상이 관찰되었다. Acanthamoeba sp. YM-4로 면역시킨 마우스의 비장세포와 myeloma 세포를 융합한 결과, 항체를 분비하는 17개 clone을 얻을 수 있었다. 생성된 단세포를 항체 중 isotyping을 실시하였는데 실험에 사용된 McAY 6, McAY 7, McAY 8, McAY 13, McAY 16 단세포군은 IgGl 항체를, McAY 10, McAY 11 단세포군은 19M 항체를 분비하였다. 생성된 단세포군 항체는 대부분이 아메바의 세포막에 있는 항원과 반응하였으며, EITB 실험결과, McAY 7 단세포군 항체는 43 kD에서 반응대가 관찰되었고, McAY 10 단세포군 항체는 55 kD과 105 kD에서 반응대가 관찰되었다. ELISA 방법을 이용한 단세포를 항체들과 여러 아메바 간의 교차반응실험 결과, McAY 7 단세포군 항체는 Acanthamoeba sp. YM-4에만 반응하였고, McAY 6과 McAY 10은 A. culbertsoni와도 반응하였다. 또한 McAY 11은 모든 아매바와 교차반응을 보였다.

• KSBB Journal
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• 제19권5호
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• pp.406-409
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• 2004

Matrix Assisted Laser Desorption ionization Time of Flight (MALDI-TOF) was used for enzymes identification related to B -1,3-glucan synthesis. Agrobacterium sp. ATCC31750 was cultivated with two stage Continuous Stirrer Tank Reactor (CSTR) and the cells were harvested and their protein profiles were analysed by two dimensional electrophoresis. The specific enzyme spot was treated with trypsin and ana lysed by MALDI-TOF to get peptide molecular weight. The peptide molecular weights were matched with Agrobacterium tumefacience's Data Base from the matrix science site, then could identify the avaliable key enzymes. In this study, we identified key metabolite of synthesis of beta-1,3-glucan, such as glucose-6-phosphate isomerase, phosphoglucomutase, B-1,3-glucan synthase and glucokinase, and we also identified uracil phosphoribocyl transferase and Ribosome recycling factor also.

• Journal of Plant Biology
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• 제39권4호
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• pp.257-263
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• 1996

Flower-inducing activity in the phloem exudata of Pharbitis cotyledons was investigated using the bioassay of Pharbitis and Lemna. By SDS-PAGE and 2-D gel electrophoresis of the phloem exudate, two polypeptides of 11 kDa and of approximately 32 kDa (pI 6.9) showing qualitative changes during the flower induction were detected. A polypeptide of approximately 20 kDa (pI 4.8) specifically labeled in vivo with [35S]methionine was found during the inductive dark period in Pharbitis cotyledon tissues. The polypeptide of the equivalent molecular mass and with the identicl pI value was also detected by in vitro translation assay. Thus, it is assumed that the 20 kDa polypeptide plays a role in the process of flower induction in Pharbitis cotyledons.

• 대한수의학회지
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• 제44권4호
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• pp.625-635
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• 2004

Trichnuris suis does not excrete eggs during larval stage as well as in particular adult stage, It is impossible to diagnose by use of fecal examination method in those periods. Therefore, serological diagnostic method can be very useful for those stages. In order to produce monoclonal antibody, specific somatic and secretory-excretory (SE) antigens of T. suis were identified and analyzed by SDS-PAGE and Western blot. Monoclonal antibody-producing hybridoma cells were cloned, which were made of popliteal lymph node of BALB/c mice immunized with a 45 kDa somatic antigen of T. suis. Five clones (1B9, 2C4, n2C5, 2D7 and 2D8) showing strong responses to T. suis antigens were selected and the isotype identified. All monoclonal antibodies were IgG1 isotype and the light chains were k chain. Established monoclonal antibodies reacted specifically to somatic and SE antigens of T. suis and did not cross-reacted to antigens of ascaris suum, trichuris vulpis, or Trichinella spiralis. The sensitivity of somatic and SE antigens against these monoclonal antibodies were significant (p<0.01) associated with those of positive and negative sera.

• 생명과학회지
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• 제16권7호
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• pp.1148-1157
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• 2006

초고온성 세균인 Thermotoga maritima로부터 ${\beta}-glucosidase$ 유전자를 클로닝한 후 대장균 숙주에서 발현시켰다. 이 효소는 salicin, arbutin, $_pNPG$과 같은 탄소원의 ${\beta}$-글루코시드 결합을 가수분해하였다. 721개의 아미노산을 암호화하는 2,166 bp의 DNA 염기서열로된 유전자이였다. 다른 ${\beta}-glucosidase$ 효소들과 단백질 유사성을 비교한 결과 glycosyl hydrolase family 3에 속하였으며 MUG-nondenaturing PAGE와 SDS-PAGE에 의해 확인된 단백질의 크기는 약 81 kDa이었다. 효소활성은 pH 7.0, $80^{\circ}C$에서 가장 높은 활성을 나타냈으며 이 효소의 아미노산 서열에 있는 두 개의 아미노산 잔기 (232번 글루탐산과 242번 아스파르트산 잔기)를 알라닌으로 치환시켜 활성이 없어지는 것으로 보아 이 두 잔기가 효소활성에 중요한 역할을 하는 것으로 추정된다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.94-100
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• 2003

We studied the effect of cadmium on chlorophylls and rubisco activation in Canavalia ensiformis L. leaves. Chlorophyll levels were reduced by 5.0 ${\mu}$M Cd. Rubisco activity at 5.0 ${\mu}$M Cd was significantly smaller than that at no treatment. Rubisco Content showed patterns of change similar to rubisco activity. These data suggest that rubisco activity was associated with an amount of rubisco protein, and that the activation and induction of rubisco is inhibited by Cd. The degree of intensity of 50 and 14.5 kD polypeptides identified as the large and small subunit of rubisco by SDS-PAGE analysis at 5.0 ${\mu}$M Cd was significantly lower than that at control, indicating Cd had a e f-fect on both subunits. Under the assumption that effects of Cd on rubisco may be r elated to rubisco activase, in addition to, its activity and content we re determined . The rubisco activase activity at 5.0 ${\mu}$M Cd was more decreased than the control. A similar change pattern was also observed in content of rubisco activase. Remarkable differences in the intensitiy of both the 45 kD and 41 kD band were found between at control and Cd-treatment. These results suggest that the change in the levels of rubisco activase leads to a subsequent alter action of rubisco levels.

• 약학회지
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• 제37권2호
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• pp.129-135
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• 1993

Serratia sp. Y-4 was isolated from soil. Many characteristics of the strain and optimum cultivation condition for protease production were investigated.,The protease from Serratia sp. Y-4 was purified and studied for the properties of the enzyme. The isolated strain was identified to the genus Serratia. The strain was cultivated in 1%-casein, 0.5%-Na$_{3}$PO$_{4}$.7H$_{2}$O, 0.1%-NaCl, 0.05%-KCI, 0.02%-MgSO$_{4}$.7H$_{2}$O, 0.02%-CaCl$_{2}$.2H$_{2}$O, 0.02%-ZnSO$_{4}$.7H$_{2}$O, 0.02%-MnCl$_{2}$.4H$_{2}$O and 0.5%-soy bean oil at pH 7.0 for 35 hrs. The enzyme was purified about 5.89 fold from the culture broth with 31.1% recovery and 19,613 u/mg through ultrafiltration, ammonium sulfate, DEAE-sephacel and Superose-12 chromatography. The purified enzyme was identified as one band by isoelectric focusing, SDS- and native-PAGE. It has maxium activity at $37^{\circ}C$ and pH 9.0. Molecular weight of it is approx. 50 kD and pl is about 6.70. Its Km value for casein was 20 mg/ml. 5 mM-EDTA, 5mM-SDS, Ag$^{+1}$, Cu$^{+2}$, Hg$^{+2}$ and Pb$^{+2}$ inhibited the enzyme.

• International Journal of Industrial Entomology and Biomaterials
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• 제26권2호
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• pp.67-73
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• 2013

Antheraea mylitta Drury is basically a crossbreeding species, as such it seems to be potentially a good material for the exploitation of heterosis. In the present study F1 hybrid of wild ecorace Laria (L) and semi-domestic Daba (D) was raised and evaluated for various quantitative traits and biochemical parameters during larval stage. Improved fecundity ($+18{\pm}1.8%$ and higher egg hatching rate ($+10.96{\pm}1.3%$) was recorded in the F1hybrid ($L{\times}D$). Biochemical parameters studied in the hemolymph, midgut and fatbody of the larva showed significantly higher (P<0.05) total proteins and carbohydrate concentration besides digestive enzyme activity. Correspondingly SDS-PAGE revealed more number of protein bands in the hemolymph sample of F1s, ranging between 29 kDa to 66 kDa compared to parental lines. The present study demonstrates the positive heterosis effect in the F1 hybrid of Laria ${\times}$ Daba. Biochemical analysis indicates that, there is possibilities of exploitation of hybrids with specific parents targeted for desirable commercial traits (silk yield and fecundity). Moreover, most of these biochemical parameters can be used as markers to analyze the genetic improvement in the tasar silkworms.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권2호
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• pp.217-221
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• 2008

Dung beetle larvae at the $3^{rd}$ instar were injected with lipopolysaccaride and inducible proteins were examined within a pI level of 3-10 and a size level by proteomics, including 1-D SDS PAGE analysis and antibacterial assay. The immune infected larvae extracts provided seven protein bands in one-dimensional electrophoresis and its antibacterial activity also checked. Hemolymph protein from immune infected larvae of the dung beetle were separated by twodimensional gel electrophoresis and compared with those from native larvae. In 2-D gel electrophoresis, we detected 63 immune infected unique and 32 up-regulated proteins, and 36 proteins that were down-regulated or not present in treated gel. Ten protein spots from unique proteins and those presented as different level of abundance in infected and native larvae were specially expressed. These differentially expressed proteins were proposed to be involved in the defense mechanism against microorganism.