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Volume 107, Issue 1

Purification and Properties of an NAD(P)-linked Formaldehyde Dehydrogenase from (Bath) Free

Abstract

Crude soluble extracts of strain Bath, grown on methane, were found to contain NAD(P)-linked formaldehyde dehydrogenase activity. Activity in the extract was lost on dialysis against phosphate buffer, but could be restored by supplementing with inactive, heat-treated extract (70 °C for 12 min). The non-dialysable, heat-sensitive component was isolated and purified, and has a molecular weight of about 115000. Sodium dodecyl sulphate gel electrophoresis of the protein suggested there were two equal subunits with molecular weights of 57000. The heat-stable fraction, which was necessary for activity of the heat-sensitive protein, was trypsin-sensitive and presumed to be a low molecular weight protein or peptide. A number of thiol compounds and other common cofactors could not replace the component present in the heat-treated soluble extract. The purified formaldehyde dehydrogenase oxidized three other aldehydes with the following values: 0·68 m (formaldehyde); 0·075 m (glyoxal); 7·0 m (glycolaldehyde); and 2·0 m (-glyceraldehyde). NAD or NADP was required for activity, with values of 0.063 and 0.155 m respectively, and could not be replaced by any of the artificial electron acceptors tested. The enzyme was heat-stable at 45 °C for at least 10 min and had temperature and pH optima of 45 °C and pH 7.2 respectively. A number of metal-binding agents and substrate analogues were not inhibitory. Thiol reagents gave varying degrees of inhibition, the most potent being -hydroxymercuribenzoate which at 1 m gave 100 % inhibition. The importance of possessing an NAD(P)-linked formaldehyde dehydrogenase, with respect to , is discussed.

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© Society for General Microbiology 1978
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1978-07-01
2024-04-26
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Purification and Properties of an NAD(P)+-linked Formaldehyde Dehydrogenase from Methylococcus capsulatus (Bath)
Microbiology 107, 19 (1978); https://doi.org/10.1099/00221287-107-1-19
/content/journal/micro/10.1099/00221287-107-1-19
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