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Purification and characterization of NADH oxidase from the archaea Sulfolobus acidocaldarius and Sulfolobus solfataricus.
Biotechnol Appl Biochem 1996; 23(1):47-54BA

Abstract

The enzyme NADH oxidase (EC 1.6.99.3) has been isolated from the two thermoacidophilic archaea Sulfolobus acidocaldarius and Sulfolobus solfataricus and characterized. In both organisms the enzyme oxidizes specifically beta-NADH in the presence of molecular oxygen and requires the presence of a flavin cofactor, showing a high specificity for FAD. A stoicheiometric amount of hydrogen peroxide to NADH is formed as the end product of the reaction, indicating that both enzymes are two-electron donors. The purified enzymes exhibit quite different molecular properties. S. acidocaldarius NADH oxidase is a monomeric protein with an estimated molecular mass of about 27 kDa, whereas S. solfataricus NADH oxidase is a dimeric protein with a molecular mass of 35 kDa per subunit; S. solfataricus NADH oxidase is purified as an FAD-containing protein, whereas S. acidocaldarius NADH oxidase does not contain a flavin molecule. Furthermore, a comparison of the N-terminal amino acid sequence shows no similarities either between the two proteins or to any other NADH oxidases. Both enzymes are essentially thermophilic. In the temperature range 20-80 degrees C, the energy of activation is almost the same for both activities, suggesting that similar energetic parameters are required. Also both oxidases display a great stability to heat. The half-life of heat inactivation is about 180 min at 90 degrees C for S. acidocaldarius NADH oxidase and 77 min at 98 degrees C for the S. solfataricus enzyme. The activity of the two enzymes is inhibited by urea and guanidine and are regulated very differently by several organic solvents.

Authors+Show Affiliations

Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8867896

Citation

Masullo, M, et al. "Purification and Characterization of NADH Oxidase From the Archaea Sulfolobus Acidocaldarius and Sulfolobus Solfataricus." Biotechnology and Applied Biochemistry, vol. 23, no. 1, 1996, pp. 47-54.
Masullo M, Raimo G, Dello Russo A, et al. Purification and characterization of NADH oxidase from the archaea Sulfolobus acidocaldarius and Sulfolobus solfataricus. Biotechnol Appl Biochem. 1996;23(1):47-54.
Masullo, M., Raimo, G., Dello Russo, A., Bocchini, V., & Bannister, J. V. (1996). Purification and characterization of NADH oxidase from the archaea Sulfolobus acidocaldarius and Sulfolobus solfataricus. Biotechnology and Applied Biochemistry, 23(1), pp. 47-54.
Masullo M, et al. Purification and Characterization of NADH Oxidase From the Archaea Sulfolobus Acidocaldarius and Sulfolobus Solfataricus. Biotechnol Appl Biochem. 1996;23(1):47-54. PubMed PMID: 8867896.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification and characterization of NADH oxidase from the archaea Sulfolobus acidocaldarius and Sulfolobus solfataricus. AU - Masullo,M, AU - Raimo,G, AU - Dello Russo,A, AU - Bocchini,V, AU - Bannister,J V, PY - 1996/2/1/pubmed PY - 1996/2/1/medline PY - 1996/2/1/entrez SP - 47 EP - 54 JF - Biotechnology and applied biochemistry JO - Biotechnol. Appl. Biochem. VL - 23 IS - 1 N2 - The enzyme NADH oxidase (EC 1.6.99.3) has been isolated from the two thermoacidophilic archaea Sulfolobus acidocaldarius and Sulfolobus solfataricus and characterized. In both organisms the enzyme oxidizes specifically beta-NADH in the presence of molecular oxygen and requires the presence of a flavin cofactor, showing a high specificity for FAD. A stoicheiometric amount of hydrogen peroxide to NADH is formed as the end product of the reaction, indicating that both enzymes are two-electron donors. The purified enzymes exhibit quite different molecular properties. S. acidocaldarius NADH oxidase is a monomeric protein with an estimated molecular mass of about 27 kDa, whereas S. solfataricus NADH oxidase is a dimeric protein with a molecular mass of 35 kDa per subunit; S. solfataricus NADH oxidase is purified as an FAD-containing protein, whereas S. acidocaldarius NADH oxidase does not contain a flavin molecule. Furthermore, a comparison of the N-terminal amino acid sequence shows no similarities either between the two proteins or to any other NADH oxidases. Both enzymes are essentially thermophilic. In the temperature range 20-80 degrees C, the energy of activation is almost the same for both activities, suggesting that similar energetic parameters are required. Also both oxidases display a great stability to heat. The half-life of heat inactivation is about 180 min at 90 degrees C for S. acidocaldarius NADH oxidase and 77 min at 98 degrees C for the S. solfataricus enzyme. The activity of the two enzymes is inhibited by urea and guanidine and are regulated very differently by several organic solvents. SN - 0885-4513 UR - https://www.unboundmedicine.com/medline/citation/8867896/Purification_and_characterization_of_NADH_oxidase_from_the_archaea_Sulfolobus_acidocaldarius_and_Sulfolobus_solfataricus_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0885-4513&date=1996&volume=23&issue=1&spage=47 DB - PRIME DP - Unbound Medicine ER -