Stabilized, NAD(P)H-dependent, soluble nitrate reductase, a proc

Details Stabilized, NAD(P)H-dependent, soluble nitrate reductase, a proc Stabilized, NAD(P)H-dependent, soluble nitrate reductase, a proc

1990-08-01

1992-12-08

Lacey, David L.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving oxidoreductase

435 37, 435188, 435189, 435184, 435191, C12Q 126, C12N 906

Patent

active

051697580

ABSTRACT:
The present invention provides a stabilized, NAD(P)H-dependent, soluble nitrate reductase of the assimilatory type, characterized by a molecular weight of about 90,000 D in the case of electrophoresis in the presence of sodium dodecyl sulphate and a residual activity after 3 weeks at 35.degree. C. of more than 60%, obtainable by preparing a suspension of the comminuted starting material in tris/tartaric acid buffer (pH 7-8.5), adding soluble polyethyleneimine thereto in such an amount that 10 to 20% of the nitrate reductase activity passes into the precipitation obtained, separating off the precipitate and working up the supernatant according to conventional biochemical methods of fractionation in the above-mentioned digestion buffer, dialyzing the enzyme solution obtained and lyophilizing in a zwitterionic buffer.

REFERENCES:
patent: 3764478 (1973-10-01), Bergmeyer
patent: 4587220 (1986-05-01), Mayambala-Mwanika et al.
patent: 4663288 (1987-05-01), Lloyd et al.
Minagawa et al. J. Biochem., vol. 91, 761-774 (1982).
Sigma Catalog, 1987, pp. 306 and 703.
Enzyme Nomenclature, 1979, Academic Press.
Downey et al, J. Bacteriology, vol. 137, pp. 105-114 (1979).
The Proceedings of Biotechnology, Band 1, 1984, Seiten 379-390; P. Monsan et al.: "Stabilization of enzyme activity".
Methods of Enzymatic Analysis, Band VII, Edition 3, 1985, Seiten 578-585: S. Taniguchi et al.: "Nitrate".

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