• Title/Summary/Keyword: purpald

Search Result 5, Processing Time 0.024 seconds

Isolation and Purification of Methyl Mercaptan Oxidase from Thiobacillus thiooxidans for Detection of Mercaptasn (머캅탄류 검출을 위한 Thiobacillus thiooxidans가 생산하는 메칠머캅탄 산화효소의 분리 및 정제)

  • 김상준;신현재;이대실;양지원
    • KSBB Journal
    • /
    • v.15 no.2
    • /
    • pp.145-149
    • /
    • 2000
  • Methyl mercaptan oxidase was isolated and purified from Thiobacillus thiooxidans KCTC2505 for the detection of mercaptans. T The produre of purification involved DEAE-Sephacel and Superose 12 column chromatographies with recovery yields of 40 a and 6.3%, and specific activity of 19.7 and 80.1 units/mg-protein, respectively. The molecular weight of purified methyl m mercaptan oxidase was determined to be 68.1 kDa by SDS-PAGE. The extract from DEAE-Sephacel column chromatography h had a high activity in oxidizing methyl mercaptan to produce formaldehyde which can be easily detected by purpald-coloring m method. Optimum temperature for activity was observed at $43^{\circ}C$. This enzyme was activated by $NH_4CI and (NH_4)_2S0_4$, and | inhibited by KCI and NaC!.

  • PDF

Isolation and Purification of Methyl Mercaptan Oxidase from Rhodococcus rhodochrous for Mercaptan Detection

  • Kim, Sang-Joon;Shin, Hyun-Jae;Kim, Yeu-Chun;Lee, Dae-Sil;Yang, Ji-Won
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.5 no.6
    • /
    • pp.465-468
    • /
    • 2000
  • Methyl mercaptan oxidase was successfully induced from Rhodococcus rhodochrous IGTS8 using methyl mercaptan gas and purified to homogeneity for the detection of mercaptans. The purification procedure involved DEAE-Sephacel and Superose 12 column chromatography with recovery yields of 85.8 and 83.3%, and a specific activity of 92.7 and 303.4 units/mg-protein, respectively. The molecular weight of purified methyl mercaptan oxidase was determined to be 64.5 kDa by SDS-PAGE. The extract from gel filtration chromatography oxidizes methyl mercaptan to produce formaldehyde, which can be easily detected by the purpald-coloring method. Optimum temperature for activity was achieved at 60$^{\circ}C$. This enzyme was inhibited by both K$_2$SO$_4$and NaCl at concentration of less than 100mM and recovered to original activity at concentration of 200mM. In the presence of methanol, the activity decreased by 33%.

  • PDF

Purification and Characterization of Methyl Mercaptan Oxidase from Thiobacillus thioparus for Mercaptan Detection

  • Lee, Hyun-Ho;Kim, Sang-Joon;Shin, Hyun-Jae;Park, Ji-Yeon;Yang, Ji-Won
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • v.7 no.6
    • /
    • pp.375-379
    • /
    • 2002
  • Methyl mercaptan oxidase was successfully induced in Thiobacillus thioparus TK-m using methyl mercaptan gas, and was purified for the detection of mercaptans. The purification procedure Involved a DEAE (diethylaminoethyl) -Sephacel, or Superose 12, column chromatography with recovery yields of 47.5 and 48.5%, and specific activities of 374 and 1240.8 units/mg-protein, respectively, The molecular weight of the purified methyl mercaptan oxidase was 66.1kDa, as determined by SDS-PAGE. The extract, from gel filtration chromatography oxidizes methyl mercaptan, producing formaldehyde, which can be easily detected by the purpald-coloring method. The optimized temperature for activity was found to be at 55$\^{C}$. This enzyme was inhibited by both NH$_4$Cl and (NH$_4$)$_2$SO$_4$, but was unaffected by either KCl or NaCl at less than 200 mM. With K$_2$SO$_4$, the activity decreased at 20 mM, but recovered at 150 mM. In the presence of methanol, full activity was maintained, but decreased in the presence of glycerin, ethanol and acetone 43, 78 and 75%, respectively.

머캅탄류 검출을 위한 Thiobacillus thioparus가 생산하는 메칠머캅탄 산화효소의 분리 및 정제

  • Kim, Sang-Jun;Sin, Hyeon-Jae;Yang, Ji-Won
    • 한국생물공학회:학술대회논문집
    • /
    • 2000.04a
    • /
    • pp.485-488
    • /
    • 2000
  • Methyl mercaptan oxidase was isolated and purified from Thiobacillus thioparus TK-m for the detection of mercaptans. The procedure of purification involved DEAE-Sephacel and Superose 12 column chromatographies with recovery yields of 47.5 and 48.5 %, and specific activity of 374 and 1240.8 units/mg-protein, respectively. The molecular weight of purified methyl mercaptan oxidase was determined to be 66.1 kDa by SDS-PAGE. Optimum temperature for activity was observed at $55\;^{circ}C$. This enzyme was activated by $(NH_4)_2SO_4$ and NaCl and inhibited by $NH_4Cl$.

  • PDF

Study on Effective Preservation of Bovine Pericardium Using Decellulariation and ${\alpha}$-galactosidase for Eliminating Xenoreactive Antigen (이종 항원 제거를 위한 무세포화와 알파-갈락토시다아제를 이용한 효과적인 우심낭 보존 방법에 관한 연구)

  • Kim, Min-Seok;Park, Cham-Jin;Kim, Soo-Hwan;Lim, Hong-Gook;Kim, Yong-Jin
    • Journal of Chest Surgery
    • /
    • v.43 no.6
    • /
    • pp.576-587
    • /
    • 2010
  • Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.