PRODUCING PULLULANASE WITH ORGANISMS HAVING A SUPERIOR CAPACITY TO ELABORATE PULLULANASE

摘要

1499340 Making pullulanase preparations A E STALEY MFG CO 17 Feb 1975 [25 Feb 1974] 6539/75 Heading C3H A method for producing pullulanase by inoculating an aqueous nutrient medium containing assimilable carbon and nitrogen sources with an aerobic, pullulanase-producing microbial organism to provide a culture thereof, incubating the culture under aerobic conditions conducive to the production of pullulanase and thereby providing production culture media thereof, and thereafter recovering a pullulanase preparation from the production culture media, is characterized in that the microbial organism (a) elaborates at least three times more pullulanase when amylopectin of a dextrose equivalent of less than 2À0 is employed as the sole carbohydrate source comparative to pullulanase elaboration in a nutrient medium wherein the major carbohydrate, on an equivalent carbohydrate weight basis, is a carbohydrate source member selected from maltose, dextrose, lactose and pullanan; (b) elaborates pullulanase at a ratio of extra-cellular pullulanase to superficially bound cell pullulanase of from 2 : 3 to less than 7 : 3whentheorganismiselaboratedinanutrient medium containing amylopectin as the sole carbohydrate source; (c) elaborates pullulanase in a nutrient medium which contains dextrose as a sole carbohydrate source; and wherein the organism is incubated in a nutrient medium for a period of time sufficient to provide a production culture medium which contains at least 350 units of pullulanase for each millilitre of production culture medium, the production culture medium containing a ratio of extracellular pullulanase to superficially bound pullulanase of from 2 : 3 to less than 7 : 3. Preferably the micro-organism used is an artificially induced Klebsiella mutant preparation containing Klebsiella pneumoniae NRRL-B-5780, Klebsiella pneumoniae NRRL-B-5783 and Klebsiella pneumoniae NRRL-B-5784.