• Journal of Ginseng Research
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• 제37권1호
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• pp.117-123
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• 2013

Polyphenol oxidase (PPO) was purified from fresh ginseng roots using acetone precipitation, carboxymethyl (CM)-Sepharose chromatography, and phenyl-Sepharose chromatography. Two isoenzymes (PPO 1 and PPO 2) were separated using an ion-exchange column with CM-Sepharose. PPO 1 was purified up to 13.2-fold with a 22.6% yield. PPO 2 bound to CM-Sepharose, eluted with NaCl, and was purified up to 22.5-fold with a 17.4% yield. PPO 2 was further chromatographed on phenyl-Sepharose. The molecular weight of the purified PPO 2 from fresh ginseng was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was about 40 kDa. The optimum temperature and pH were $20^{\circ}C$ and 7.0, respectively, using catechol as a substrate. Pyrogallol showed the highest substrate specificity. The effect of a PPO inhibitor showed that its activity increased slightly in the presence of a low concentration of citric acid. High concentrations of acidic compounds and sulfite agents significantly inhibited purified ginseng PPO 2.

• Preventive Nutrition and Food Science
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• 제7권2호
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• pp.184-187
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• 2002

Blood products from slaughterhouses that are not hygienically prepared for disposal or food consumption pose a human health hazard. Gamma irradiation is an effective method for sterilization of blood products, but may introduce changes in the molecular characteristics of proteins. This study evaluated the effects of irradiation on animal plasma proteins. Bovine and porcine blood was obtained from a slaughterhouse and the plasma proteins purified and lyophilized. The secondary structure and molecular weight distribution of the plasma protein solutions and powders were examined after ${\gamma}$-irradiation at 1, 5, 7 and 10 kGy. Gamma-irradiation affected the molecular properties of the protein solutions, but not the protein powders. Circular dichroism and sodium dodecyl sulfate-polyacrylamide gel electrophoresis studies showed that increased doses of ${\gamma}$-irradiation decrease the ordered structure of plasma proteins in solution, and cause initial fragmentation of the polypeptide chains and subsequent aggregation.

• 한국식품과학회지
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• 제48권6호
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• pp.542-547
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• 2016

The thermal properties of ramie leaf ${\beta}$-amylase (RBA) were examined to develop a novel process for enzyme purification. The thermostability of RBA extract prepared from ramie leaf powder was examined at various temperatures. RBA activity decreased slightly, whereas other carbohydrate-active enzymes, such as $\small{D}$-enzyme, were rapidly inactivated during 30 min incubation at $60^{\circ}C$. When the heat-treated extract was incubated with various substrates, maltose was produced exclusively as the major product, whereas the untreated crude extract produced maltose and other maltooligosaccharides. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, fewer protein bands were observed for the heat-treated extract than the untreated extract, indicating that the thermostable RBA was partially purified and other thermolabile enzymes were eliminated. Thus, the treatment of the RBA extract at $60^{\circ}C$ for 30 min resulted in 5.4-fold purification with a recovery yield of 90%.

• 한국축산식품학회지
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• 제21권3호
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• pp.246-255
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• 2001

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of muscles extracted with distilled water, saline solution, SDS or Trition X-100 showed simular protein patterns between Hanwoo and Holstein meat, indicating that SDS-PAGE technique may not be useful for the identification between Hanwoo and Holstein meat. Lectine blot analysis of muscle extracted with distilled water demonstrated that Hanwoo and Holstein meat had similar affinities for concanavalin A (Con A), ricinus communis agglutinin (RCA-120), ulex europaeus agglutinin (UEA-1) or peanut agglutinin (PNA) lectins. However, approximately 32.1 kDa component of Hanwoo meat showed high affinity for dolichos biflorus agglutinin (DBA) lectin. On the contrary, high molecular weight components of Holstein meat had the specific affinity for wheat germ agglutinin (WGA) lectin. Hanwoo meat-specific components were observed by lectin staining of heat-denatured meat at 100$^{\circ}C$ for 30 sec. Also, the component of heat-denatured meat at 100$^{\circ}C$ for 30 sec, which was slightly smaller than Hanwoo meat-specific component, was concentrated specifically in Holstein meat.

• 대한수의학회지
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• 제25권2호
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• pp.149-153
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• 1985

Salmonella pullorum strain W was serially passaged on the brain heart infusion agar containing acrdine orange(AO) as a concentration of 100 mcg/ml. S. pullorum AO60 and S. pullorum AO150, which were subcultured 60 and 150 passages on AO media, were examined for permeability barrier function of the cell wall. AO60 and AO150 were appeared to be decreased in susceptibility against hydrophobic substances such as crystal violet, chloramphenicol and rifamycin, which might be resulted from the changes of permeability barrier function of the cell wall. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis of bacterial protein, the protein profiles of AO6O and AO150 didn't differ significantly from W, but increased amount of the band of MW 140,000-145,000 was confirmed. And [G+C] contents of DNA in AO60 and A0150 were decreased than that of W.

• Archives of Pharmacal Research
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• 제19권6호
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• pp.502-506
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• 1996

Protein X is one of the subunits of pyruvate dehydrogenase complex. The biological role of this protein has not been fully elucidated, mainly because of the difficulty in its dissociation from the tightly bound dihydrolipoamide acetyltransferase-protein X subcomplex. We have found that the detachment of protein X from acetyltransferase subunit can be easily accomplished by the cycles of freezing and thawing proces. Several lines of evidence including sodium dodecyl sulfate-polyacrylamide gel electrophoresis, N-terminal amino acid sequence analysis and acetylation with $[2^{14}C]$ pyruvate confirmed that the purified protein is protein X. The purified intact form of protein X was acetylated by $[2^{14}C]$ pyruvate in the presence of py-ruvate dehydrogenase subunit.The acetylation efficiency of this protein was lower than that of acetyltransferase and was not affected by the presence of acetyltransferase.

• KSBB Journal
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• 제5권1호
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• pp.19-23
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• 1990

콜레스테롤 에스테라아제를 생산하는 미생물 strain CES506 균주를 토양으로부터 분리하여 Aeromonas속의 한 세균으로 동정하였다. 이 균주는 배양액 1ml당 0.023 단위의 콜레스테롤 에스테라아제를 생산하였다. 이 균주가 생산하는 본 효소를 균주 배양액으로부터 약 370배, 24%의 수율로 균질한 상태의 단백질로 정제하였다. 본 효소의 분자량은 SDS-PAGE로 산출한 결과 64,000으로 계산되었다. 본 효소는 cholesteryl-palmitate에 대해 높은 기질 특이성을 나타내었으며 cholesterylpalmitate의 가수분해에 대한 Km값은 0.15mM이었다.

• Journal of Applied Biological Chemistry
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• 제55권3호
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• pp.173-178
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• 2012

충치 형성 과정에 중요한 불용성 글루칸의 합성과 관련된 sucrose-glucan glucosyltransferase (Gtf)를 Streptococcus mutans Ingbritt 의 배양액에서 염석과, Sephadex G-150, CM-Sephadex, DEAE-Sephadex column을 통과시켜 정제하였다. 회수율 6.3%였으며, 정제율 13배였다. sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)에서 이 효소의 분자량은 66 kDa이며, 최적 pH와 온도는 각각 6.0과 $40^{\circ}C$이였다. 이 효소는 0.1 mM $Hg^{2+}$, $Cu^{2+}$, $Al^{3+}$에 의하여 22-59% 저해되었다. 이 효소는 2 mM 자이리톨에 의하여 40% 저해 받았으며, 0.05% 수용성 키토산, 글리콜 키토산, 글리콜 키틴에 의하여 35-45% 저해되었다. 키틴 유도체가 Gtf 활성을 억제한다는 보고는 본 연구가 최초이며, 이는 키틴 유도체를 구강청결제의 소재로 활용할 수 있음을 제시한다.

• 미생물학회지
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• 제28권2호
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• pp.128-133
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• 1990

고등한 진핵생물에서 nucleosome 구성에 중요한 역할을 하는 histone 단백질이 하등한 진핵생물에서도 존재하는지의 여부를 알아보기 위하여 실험을 하였다. 하등한 균류에 속하는 Rhizopus 속의 5종(R. acidus, R. arrhizus, R. formosaensis. R R. japoηicus, R. oryzae)을 재료로 하여 15% polyacrylamide acid-urea gel과 18% P이yacηlam ide SDS-gel을 이용하여 전기영동하였다. 분자량을 비교하기 위하여 marker protein으로 Cytochrome C와 함께 15% polyacrylamide SDS-gel을 이용하였으며 모든 gel에서 standard로써 calf thymus histone을 사용하여 electrophoreticmobility를 비교하였고 호 한 속내에서의 종간의 차이가 있는지를 설험하였다 실험결과는 한 속내에서 종에 따른 차이로 볼 수 없었으며 standard인 calf thymus histone HI. H2A, H2B, H3, H4와 유사한 electromobility흘 갖는 protein band를 확인할 수 있었으며 HI histone은 t성확하게 일치되지가않았다.따라서 하등한균류인Rhizopµs에서도고등 진핵생불과유사한단백질이 존재함을 주정할 수가 있었다.

• 한국미생물·생명공학회지
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• 제17권5호
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• pp.440-446
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• 1989

Penicillium sp. CB-20의 polygalacturonase 생성을 위한 최적조건은 탄소원으로 펙틴을 사용하여 16 시간 배양시 최대 활성을 나타내었으며, Sephadex G-25, G-75 및 G-150을 사용한 gel filtration과 DEAE-cellulose, DEAE-Sephadex A-50에 의 한 ion exchange chromatography를 통하여 이 효소를 약 30배 가량 정제할 수 있었고, 수율은 2.31%였다. 정제효소는 polyacrylamide gel electrophoresis에 의하여 단일 밴드로 확인 되었으며, 분자량은 SDS PAGE에 의하여 21,000 정도로 측정되었다. 효소의 결정구조는 마름모꼴을 형성하고 있었으며 아미노산 조성은 17종류로써 glutamic acid, glycine, hlstidine의 함량이 비교적 많았다.