New Feed Enzyme Development - The Poultry Federation

September 2009, ChemGen Corp.First the purified β‐1,3‐glucanase was characterized to establish the purity and specificity of the enzyme.The purified enzyme was not 100% pure, but highly purified and free of other glucanases (amylase andcellulase) . The purity is analyzed in Figure 12 and specificity in Figure 13.Figure 12 ‐ Purified β‐1,3‐glucanase on SDS‐PAGE gelPurified β‐1,3‐glucanase from Hemicell®production host strain analyzed on SDSpage gel (Lanes 2 and 3) compared toBio‐Rad standard broad range proteinmolecular weight markers (Lane 1).Marked bands on the left are molecularweights in kilo Daltons.Lane 2, normal loading, Lane 3, heavyloading to check purity. The β‐1,3‐glucanase is marked with an arrow.Figure 13‐ Verification of the specificity of purified β‐1,3‐glucanase with defined β‐1,3‐glucans, CMPand ScleroglucanCarbohydrate gel as described in Figure 7modified with higher % gel. CMP and wasobtained from Megazyme and Scleroglucanfrom Cargill.Carbohydrate Gel lanes1‐glucose (dp 1)2‐ maltose (dp 2)3‐maltotriose( dp 3)4‐ maltodextrin (dp 4‐10)(dp 8‐10 not resolved )5‐ CMP (contaminated with glucose, noenzyme)6,8 –CMP (carboxymethyl pachyman)digest, two levels loading7,9 – scleroglucan digest, two levelsloading32% acrylamine gel, 2 hours, 200 VLanes 6‐9 digested withβ‐1,3‐glucanase before ANTS reactionThe purified β‐1,3‐glucanase was able to digest two known available polymers with β‐1,3‐glucanbackbone, carboxymethy pachyman and sclerogucan. Further the gels show that that the enzyme hadD.M. Anderson and H. Hsiao, New Feed Enzymes Page 17