Investigation of the H2-oxidizing activities of Alcaligenes eutrophus H16 membranes with artificial electron acceptors, respiratory inhibitors and redox-spectroscopic procedures.
Membrane particles, prepared from cells of Alcaligenes eutrophus H16 by lysozyme treatment and 100 000 X g centrifugation, catalyzed a H2-dependent reduction of methylene blue, menadione, 2,6-dichlorophenol-indophenol (DCPIP), and O2. While the reaction with methylene blue was not altered by 2-n-heptyl-4-hydroxyquinoline-N-oxide (HQNO), the H2-dependent reductions of menadione, DCPIP and O2 were strongly inhibited, indicating that in these reaction components of the respiratory chain other than the membrane-bound hydrogenase were involved. The effect of pentane extraction of membranes on the H2-dependent reductions of methylene blue and menadione were different from those of DCPIP and O2. This suggested that ubiquinone might not be involved in the pathway of the electrons from H2 to methylene blue or menadione, while it might be involved in the pathway to DCPIP and O2. Because the H2-dependent reduction of menadione is sensitive to HQNO, it follows that HQNO might bind to a site upstream of ubiquinone. Further evidence for this hypothesis came from a new technique to record UV and visible redox-difference spectra of membranes under the conditions of a steady-state electron flow. HQNO did not increase the reduction level of ubiquinone relative to the cytochromes. Neither HQNO nor menadione had any influence on the redox difference patterns of the cytochromes as determined with low temperature and room temperature spectroscopy.
D015086:2,6-Dichloroindophenol / Q000378:metabolism; D000421:Alcaligenes / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D003580:Cytochromes / Q000378:metabolism; D004579:Electron Transport; D006859:Hydrogen / Q000378:metabolism*; D006864:Hydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006912:Hydroxyquinolines / Q000494:pharmacology; D008751:Methylene Blue / Q000378:metabolism; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D013053:Spectrophotometry; D014451:Ubiquinone / Q000378:metabolism; D014812:Vitamin K / Q000378:metabolism
D003580:Cytochromes; D006912:Hydroxyquinolines; D014812:Vitamin K; D014451:Ubiquinone; C001333:2-(n-heptyl)-4-hydroxyquinoline N-oxide; D006859:Hydrogen; D015086:2,6-Dichloroindophenol; D006864:Hydrogenase; D010100:Oxygen; D008751:Methylene Blue