caa a22 4500 46909317X CHVBK 20180323133023.0 cr unu---uuuuu 170328e19921001xx s 000 0 eng 10.1007/BF00582580 doi (NATIONALLICENCE)springer-10.1007/BF00582580 Purification and characterization of an NAD+-linked formaldehyde dehydrogenase from the facultative RuMP cycle methylotroph Arthrobacter P1 [Elektronische Daten] [Margaret Attwood, Nico Arfman, Ruud Weusthuis, Lubbert Dijkhuizen] WhenArthrobacter P1 is grown on choline, betaine, dimethylglycine or sarcosine, an NAD+-dependent formaldehyde dehydrogenase is induced. This formaldehyde dehydrogenase has been purified using ammonium sulphate fractionation, anion exchange- and hydrophobic interaction chromatography. The molecular mass of the native enzyme was 115 kDa±10 kDa. Gel electrophoresis in the presence of sodium dodecyl sulphate indicated that the molecular mass of the subunit was 56 kDa±3 kDa, which is consistent with a dimeric enzyme structure. After ammonium sulphate fractionation the partially purified enzyme required the addition of a reducing reagent in the assay mixture for maximum activity. The enzyme was highly specific for its substrates and the Km values were 0.10 and 0.80 mM for formaldehyde and NAD+, respectively. The enzyme was heat-stable at 50° C for at least 10 min and showed a broad pH optimum of 8.1 to 8.5. The addition of some metal-binding compounds and thiol reagents inhibited the enzyme activity. Kluwer Academic Publishers, 1992 Arthrobacter P1 nationallicence choline nationallicence formaldehyde dehydrogenase (NAD+-dependent) (EC 1.2.1.-) nationallicence methylamine nationallicence RuMP : Ribulose monophosphate nationallicence Attwood Margaret Department of Microbiology, University of Sheffield, UK aut Arfman Nico Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands aut Weusthuis Ruud Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands aut Dijkhuizen Lubbert Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands aut Antonie van Leeuwenhoek Kluwer Academic Publishers 62/3(1992-10-01), 201-207 0003-6072 62:3<201 1992 62 10482 https://doi.org/10.1007/BF00582580 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00582580 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Attwood Margaret Department of Microbiology, University of Sheffield, UK aut NATIONALLICENCE 700 1- Arfman Nico Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands aut NATIONALLICENCE 700 1- Weusthuis Ruud Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands aut NATIONALLICENCE 700 1- Dijkhuizen Lubbert Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands aut NATIONALLICENCE 773 0- Antonie van Leeuwenhoek Kluwer Academic Publishers 62/3(1992-10-01), 201-207 0003-6072 62:3<201 1992 62 10482 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer