Patent ID: 8119124

Claim:
A debriding composition obtained from bromelain, the debriding composition comprising a plurality of different proteolytic enzymes having molecular weights of about 23 kDa as determined by size exclusion chromatography, wherein the plurality of different proteolytic enzymes comprises: (a) stem bromelain comprising SEQ ID. No. 1; (b) ananain comprising SEQ ID No. 2; (c) stem bromelain comprising SEQ ID No. 3; and (d) cysteine proteinase precursor comprising SEQ ID No. 4, said debriding composition comprising bromelain inhibitors in an amount of less than 10% w/w of protein content of the debriding composition, wherein the debriding composition is obtained by the steps of: (a) suspending bromelain with an acidic solution optionally comprising an anti-oxidant, the acidic solution having a pH in the range from about 2.4 to about 4; (b) adjusting the suspension of (a) to a pH in the range from about 2.4 to about 4; (c) adding a filter aid comprising silica to the suspension of (b); (d) filtering the suspension of (c) to remove insoluble components; (e) adding to the filtered solution of (d) ammonium sulfate salt to yield saturation of ammonium sulfate in the range from about 40% to about 50%; (f) adjusting the suspension of (e) to a pH from about 2.5 to about 4; (g) incubating the suspension of (f) at 3° C.-10° C.; (h) centrifuging the suspension of (g) to yield an ammonium sulfate precipitate; (i) dissolving the ammonium sulfate precipitate in an acidic solution optionally comprising an anti-oxidant having a pH in the range from about 2.4 to about 4; (j) filtering the solution of (i) so that proteolytic enzymes having molecular weights in excess of about 10 kDa are retained; and (k) lyophilizing the retained solution of (j) to obtain the debriding composition.