Purification and characterization of an alcohol dehydrogenase from the Antarctic psychrophile Moraxella sp. TAE123

• Published in: European Journal of Biochemistry
• Main Authors: Tsigos, Iason, Velonia, Kelly, Smonou, Ioulia, Bouriotis, Vassilis
• Format: Article in Journal/Newspaper
• Language: English
• Published: Wiley 1998
• Subjects: Antarctic; The Antarctic; Antarc*
• Online Access: http://dx.doi.org/10.1046/j.1432-1327.1998.2540356.x; https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1046%2Fj.1432-1327.1998.2540356.x; https://febs.onlinelibrary.wiley.com/doi/pdf/10.1046/j.1432-1327.1998.2540356.x

An NAD + ‐dependent alcohol dehydrogenase (ADH) of the Antarctic psychrophile Moraxella sp. TAE123 was purified to homogeneity with an overall yield of 16.7 % and further characterized. The native enzyme had an apparent molecular mass of 240 kDa and consisted of four identical 52‐kDa subunits. The pI of the enzyme was determined to be 5.5, while its optimum pH is 7.5. The enzyme contained 1 zinc atom/subunit and exhibited a remarkable thermal lability. Moraxella sp. TAE123 ADH exhibited a wide range of substrate specificity similar to its mammalian counterparts and in contrast to other microbial ADHs. It oxidized mainly primary and secondary aliphatic alcohols. The highest reaction rate was observed when ethanol was used as substrate. A gradual decrease in rate was observed by increasing the length and branching of the carbon chain of the alcohol. This enzyme oxidized effectively large bulky alcohols, such as diphenylmethanol. Reduction of aldehydes and ketones was also observed. N‐terminal amino acid sequence analysis of the enzyme did not reveal any similarity with the amino termini of all other ADHs, while an unexpected significant similarity was observed with the amino terminal sequence of four prokaryotic aldehyde dehydrogenases.