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Alanine dehydrogenase of the N2-fixing blue-green alga, Anabaena cylindrica

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Abstract

The l-alanine dehydrogenase (ADH) of Anabaena cylindrica has been purified 700-fold. It has a molecular weight of approximately 270000, has 6 sub-units, each of molecular weight approximately 43000, and shows activity both in the aminating and deaminating directions. The enzyme is NADH/NAD+ specific and oxaloacetate can partially substitute for pyruvate. The K app m for NAD+ is 14 μM and 60 μM at low and high NAD+ concentrations, respectively. The K app m for l-alanine is 0.4 mM, that for pyruvate is 0.11 mM, and that for oxaloacetate is 3.0 mM. The K app m for NH +4 varies from 8–133 mM depending on the pH, being lowest at high pH levels (pH 8.7 or above). Alanine, serine and glycine inhibit ADH activity in the aminating direction. The enzyme is active both in heterocysts and vegetative cells and activity is higher in nitrogen-starved cultures than in N2-fixing cultures. The data suggest that although alanine is formed by the aminating activity of ADH, entry of newly fixed ammonia into organic combination does not occur primarily via ADH in N2-fixing cultures of A. cylindrica. Ammonia assimilation via ADH may be important in cultures with an excess of available nitrogen. The deaminating activity of the enzyme may be important under conditions of nitrogen-deficiency.

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Abbreviations

ADH:

alanine dehydrogenase

DEAE:

diethylamino ethyl cellulose

EDTA:

ethylenediamine tetraacetic acid

GDH:

glutamic dehydrogenase

GS:

glutamine synthetase

GOT:

aspartate-glutamate aminotransferase

NAD+ :

nicotinamide adenine dinucleotide

NADH:

reduced nicotinamide adenine dinucleotide

NADP+ :

nicotinamide adenine dinucleotide phosphate

NADPH:

reduced nicotinamide adenine dinucleotide phosphate

SDS:

sodium dodecyl sulphate

Tris:

tris(hydroxymethyl) aminomethane

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  1. Department of Biological Sciences, University of Dundee, DD1 4HN, Dundee, Scotland

    P. Rowell & W. D. P. Stewart

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  1. P. Rowell
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Rowell, P., Stewart, W.D.P. Alanine dehydrogenase of the N2-fixing blue-green alga, Anabaena cylindrica . Arch. Microbiol. 107, 115–124 (1976). https://doi.org/10.1007/BF00446830

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  • DOI: https://doi.org/10.1007/BF00446830

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