Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving oxidoreductase
Patent
1979-05-22
1982-08-03
Warden, Robert J.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving oxidoreductase
435172, 435188, 435190, 435253, 435832, C12Q 132, C12N 1500, C12N 904, C12N 120
Patent
active
043428274
ABSTRACT:
Glycerol dehydrogenase enzymes having exeptionally good thermal stability are produced by culturing novel strains of Bacillus stearothermophilus. Procedures for deriving and identifying suitable strains are described. The strains are grown in conventional culture media, preferably containing 0.05 to 4.0%, especially 0.1 to 1.0%, by weight of glycerol or a glycerol analogue at 40.degree.-65.degree. C. and pH 5 to 8. The enzyme is isolated by conventional cell disruption and separation techniques, and typically has a molcular weight of 240,000.+-.30,000, composed of four similar sub-units, and a specific activity of greater than 5 Units per mg protein at 30.degree. C. by the modified assay described. They may be stored as aqueous solutions or a freeze dried solids.
The enzymes may be used for assay of serum triglycerides by conventional assay methods, but preferably by the nictotinamide adenine dinucleotide spectrophotometric assay at a pH of 7 to 8.8. The pH is preferably controlled by an amine, especially triethanolamine/HCl, buffer.
REFERENCES:
Sidney P. Colowick and Nathan O. Kaplan, Methods in Enzymology, vol. 1, Academic Press Inc., Publisher, pp. 397-400; 1955.
Thomas E. Bowman, Enzyme Handbook, vol. 1, p. 29, 1969.
R. E. Buchanan and N. E. Gibbons, Co-Editors, Berzey's Manual of Determinative Bacteriology, The William and William Company, pp. 539-540; 1974.
Atkinson Anthony
Bruton Christopher J.
Comer Michael J.
Sharp Richard J.
Public Health Laboratory Service, Center for Applied Microbiolog
Warden Robert J.
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