Dextranα-1,2 Debranching Enzyme fromFlavobacteriumsp. M-73: Its Production and Purification

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Abstract
A new type of dextranase, dextran α-1,2 debranching enzyme, was detected in the culture fluid of a gram negative bacterium. This dextranase producing microorganism had characteristics of the genus Flavobacterium, and was tentatively named M-73. The optimum conditions for enzyme production were studied. The debranching enzyme was concentrated from the culture supernatant, treated at 55°C to inactivate the concomitant dextranase activity, and purified by column chromatography on DEAE-cellulose. Further purification was performed by biospecific adsorption chromatography using Sephadex G-150, from which the enzyme was eluted with a solution of clinical dextran, and by Polyacrylamide preparative gel electrophoresis. The purified enzyme was electrophoretically homogeneous and had a molecular weight of approximately 125,000. About a 2,700-fold increase in specific activity was achieved (66 units per milligram of protein) with a yield of 14.2%.

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