• Applied Biological Chemistry
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• 제31권4호
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• pp.382-386
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• 1988

Chemically defined medium을 사용하여 Bacillus sp. $T_2-3$으로 균체외단백질을 생산한 후 이들의 각종 물리화학적 성질을 조사 하였다. 균체외단백질은 전기영동과 gel 여과 결과로 분자량이 약 49,000이 되는 비슷한 2종의 단백질로 구성되어 있었다. 최대흡수파장은 230nm 부근이었으며 aspartic acid를 가장 많이 함유하고 있었다. 또한 균체외단백질의 물에대한 용해도는 55.8%, 0.4% NaOH에 대한 용해도는 28.4%로 albumin과 glutelin계통의 단백질로 생각되었다.

• Fisheries and Aquatic Sciences
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• 제2권2호
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• pp.218-225
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• 1999

A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.129-136
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• 1997

Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.570-577
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• 2003

The importance of cornified envelope (CE)-bound lipids to epidermal barrier function is increasingly being recognized. In the present study, we intentionally damaged the cornified layer of hairless mice by ultraviolet irradiation and sodium dodecyl sulfate (SDS) treatment, and assessed the changes in epidermal barrier function by measuring Trans Epidermal Water Loss (TEWL). We also measured changes in the amount of CE-bound lipids using thin layer chromatography (TLC). The results showed that both treatments increased TEWL and decreased CE-bound lipids (omega-hydroxy cerami de and omega-hydroxy acid). In addition, investigation of the chronological changes in TEWL revealed an inverse relationship between TEWL and CE-bound lipids, and a correlation between CE-bound lipids and epidermal barrier function. We then measured the amount of CE-bound lipids in the cheek and the medial side of the upper arm in humans. The results showed that because the cheek receives external stimulation on a daily basis, the amount of CE-bound lipids was significantly lower, while the level of TEWL was higher. These observations, together with those from the animal study, indicate that CE-bound lipids are related to epidermal barrier function.

• Journal of the Korean Wood Science and Technology
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• 제46권4호
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• pp.415-422
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• 2018

Terpenoids have a wide range of biological functions and have extensive applications in the pharmaceutical, cosmetic, and flavoring industry. The white-rot fungus, Polyporus brumalis, is able to synthesize terpenoids via terpene synthase, which catalyzes an important step that forms a large variety of sesquiterpene products from farnesyl pyrophosphate (FPP). To improve the production of sesquiterpenes, the terpene synthase gene was isolated from Polyporus brumalis and was heterologously transformed into a Pichia pastoris strain. The open reading frame of the isolated gene (approximately 1.2 kb) was inserted into Pichia pastoris to obtain a recombinant enzyme. Five transformants were obtained and the expression of terpene synthase was analyzed at the transcript level by reverse transcription PCR (polymerase chain reaction) and at the protein level by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). Expression of the terpene synthase gene product was elevated in the transformants and as expected the molecular weight of the protein was approximately 45 kDa. These recombinant enzymes have potential practical applications and future studies should focus on their functional characterization.

• ALGAE
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• 제27권1호
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• pp.55-62
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• 2012

A novel lectin specific to N-acetyl-D-galactosamine as well as N-acetyl-D-glucosamine was isolated from Bryopsis plumosa and named as BPL-4. Sodium dodecyl sulfate polyacrylamide gel electrophorese (SDS-PAGE) and matrix-assisted laser desorption / ionization-time of flight (MALDI-TOF) mass spectrometry data showed that this lectin was a monomeric protein with molecular weight 12.9 kDa. The N-terminal amino acid sequences of the lectin were determined by Edman degradation and the full cDNA sequence encoding this lectin was obtained using the degenerate primers designed from the amino acid sequence. The size of the cDNA was 414 bp containing single open reading frame (ORF) encoding the lectin precursor. The homology analysis showed that this lectin might belong to H lectin group. BPL-4 showed high sequence similarity (60.6%) to BPL-3, which is a previously reported lectin from the same species. The comparative analysis on the lectin's primary structure showed two conserved domains including one possible active domain of H lectin group.

• Archives of Pharmacal Research
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• 제21권3호
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• pp.291-297
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• 1998

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.360-367
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• 1990

토양으로부터 rutin을 유일한 탄소원으로 생육하는 세균을 분리하여 그 중 rutin 가수분해활성이 높은 MT-57균을 Arthrobacter sp.로 동정하였다. Arthrobacter sp.MT-57에서 수 종류의 rutin 가수분해효소가 확인되었으며 이들 중 rutinosidase를 전기영동상 단일한 효소 단백질까지 정제하였다. Rutinosidase으 분자량은 SDS page와 gel filtration으로 부터 42,000의 단량체로 추정 되었다. 효소반응의 최적 pH와 온도는 pH 7.5와 $45^{\circ}C$였으며, 20분간 열처리하였을 때 $50^{\circ}C$까지 안정하였다.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2010년도 춘계학술발표대회
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• pp.57.2-57.2
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• 2010

대표적인 투명 전극 재료indium tin oxide(ITO)의 경우, 우수한 투과성과 낮은 면저항을 기반으로 차세대 디스플레이용 전극으로 각광 받고 있다. 하지만 제조 단가가 높으며 brittle 하여 유연 디스플레이에 적용이 어려우며 대면적 제조가 어렵다는 단점이 있어 이를 대체할 수 있는 새로운 물질이 필요한 실정이다. 대표적인 후보 물질로는 탄소 육각형이 서로 연결된 관 형태인 탄소나노튜브가 있으며 뛰어난 전기 전도도와 물리적 특성을 투명 전극에 적용하기 위한 연구가 활발히 진행 중이다. 본 연구에서는 탄소나노튜브 투명 전극 제조 시 잔여 분산제 제거 및 doping의 효과를 위해 수행되는 산처리 공정을 하지 않고 투명 전극의 특성을 향상 시키는 연구를 진행하였다. 제작된 박막에 압력을 인가하여 탄소나노튜브 네트워킹의 향상과 두께의 감소를 얻을 수 있었다. 실험에 사용된 탄소나노튜브는 아크 방전 공정으로 합성된 2nm의 single wall 탄소나노튜브이며 이를 분산제인 sodium dodecyl sulfate(SDS)에 분산하여 용액형태로 제작하여 사용하였다. 분산제를 제거하기 위해 탈이온수를 사용하였으며 고분자 mold를 사용하여 압력을 인가하여 그에 따른 전기적, 광학적 변화를 관찰하였다. 제조된SWCNT 박막은 four point probe measurement를 이용하여 sheet resistance를 측정하였고 UV-vis를 이용하여 투과도와 반사도 등의 광학적 특성을 측정하였다. 박막의 표면은 field emission scanning electron microscope (FESEM)과 Atomic force microscope(AFM)를 이용하여 관찰하였다.

• Journal of Microbiology and Biotechnology
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• 제22권1호
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• pp.133-140
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• 2012

A novel cry2Ab gene was cloned and sequenced from the indigenous isolate of Bacillus thuringiensis subsp. kurstaki. This gene was designated as cry2Ab25 and its sequence revealed an open reading frame of 1,902 bp encoding a 633 aa protein with calculated molecular mass of 70 kDa and pI value of 8.98. The amino acid sequence of the Cry2Ab25 protein was compared with previously known Cry2Ab toxins, and the phylogenetic relationships among them were determined. The deduced amino acid sequence of the Cry2Ab25 protein showed 99% homology to the known Cry2Ab proteins, except for Cry2Ab10 and Cry2Ab12 with 97% homology, and a variation in one amino acid residue in comparison with all known Cry2Ab proteins. The cry2Ab25 gene was expressed in Escherichia coli BL21(DE3) cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the Cry2Ab25 protein is about 70 kDa. The toxin expressed in BL21(DE3) exhibited high toxicity against Malacosoma neustria and Rhagoletis cerasi with 73% and 75% mortality after 5 days of treatment, respectively.