Glucanase production and methods of using the same

Abstract

Methods and compositions are described for producing a glucanase in transgenic plants and then incorporating parts of the transgenic plants in animal feed. The feed glucanase enzyme displays activity across a broad pH range, and tolerance to temperatures that are often encountered during the process of preparing animal feeds. Producing the enzyme in the transgenic plant enhances the thermal stability of the enzyme.

What is claimed is: 
     
         1 . A transgenic plant, part or seed thereof comprising at least one synthetic nucleic acid encoding a glucanase and at least one synthetic polynucleotide producing a diagnostic amplicon for identifying event 4588.652, wherein the at least one nucleic acid comprises a sequence with at least 90% identity to the sequence set forth in SEQ ID NO: 8, and the at least one synthetic polynucleotide comprises a sequence with at least 90% identity to the sequence selected from the group consisting of SEQ ID NOS: 29-31. 
     
     
         2 . The transgenic plant or part thereof of  claim 1 , wherein the glucanase is capable of degrading one or more polysaccharides selected from the group consisting of beta-glucan, cellulose, cellobiose, pNP-D-glucopyranoside and xylan. 
     
     
         3 . The transgenic plant or part thereof of  claim 1 , wherein the glucanase is active upon expression in the plant and exposure to a pH in the range from 5.0 to 10.0. 
     
     
         4 . The transgenic plant or part thereof of  claim 1 , wherein the glucanase is active upon expression in the plant and exposure to a temperature in the range from 25° C. to 130° C. 
     
     
         5 . The transgenic plant or part thereof of  claim 1 , wherein the glucanase activity has improved stability upon expression in the plant compared to the activity of a glucanase having an identical amino acid sequence and expressed in a bacterial cell. 
     
     
         6 . The transgenic plant or part thereof of  claim 1 , wherein a plant is selected from the group consisting of: wheat, maize, barley, and sorghum. 
     
     
         7 . An animal feedstock comprising the transgenic plant, part or seed thereof of  claim 1 . 
     
     
         8 . The animal feedstock of  claim 7  further comprising a feed supplement. 
     
     
         9 . The animal feedstock of  claim 8 , wherein the feed supplement is plant material selected from the group consisting of: a non-transgenic plant, another transgenic, mutant and engineered plant. 
     
     
         10 . The animal feedstock of  claim 8 , wherein the feed supplement comprises one or more exogenous enzymes selected from the group consisting of: a hydrolytic enzyme selected from the group consisting of: xylanase, endoglucanase, cellulase, protease, phytase, amylase and mannanase. 
     
     
         11 . The animal feedstock of  claim 8 , wherein the feed supplement comprises at least one component selected from the group consisting of: corn meal, corn pellets, wheat meal, wheat pellets, wheat grain, barley grain, barley pellets, soybean meal, soybean oilcake, sorghum grain and sorghum pellets. 
     
     
         12 . The animal feedstock of  claim 8 , wherein the feed supplement comprises at least one component selected from the group consisting of: soluble solids, fat and vermiculite, limestone, plain salt, DL-methionine, L-lysine, L-threonine, COBAN®, vitamin premix, dicalcium phosphate, selenium premix, choline chloride, sodium chloride, and mineral premix. 
     
     
         13 . A method of producing an animal feedstock comprising mixing the transgenic plant, part or seed thereof of  claim 1  with other plant material to form a mixture. 
     
     
         14 . The method of  claim 13 , wherein the method further comprises pelletizing the mixture. 
     
     
         15 . A method of increasing metabolizable energy of a diet, wherein the method comprises mixing the transgenic plant, part, or seed thereof of  claim 1  with a feed ingredient. 
     
     
         16 . The method of  15 , wherein the feed ingredient comprises at least one component selected from the group consisting of: corn meal, corn pellets, wheat meal, wheat pellets, wheat grain, wheat middlings, barley grain, barley pellets, soybean meal, soy hulls, dried distillers grain, soybean oilcake, sorghum grain and sorghum pellets. 
     
     
         17 . The method of  claim 15 , wherein the feed ingredient comprises at least one component selected from the group consisting of: soluble solids, fat and vermiculite, limestone, plain salt, DL-methionine, L-lysine, L-threonine, COBAN®, vitamin premix, dicalcium phosphate, selenium premix, choline chloride, sodium chloride, mineral premix, and one or more exogenous enzymes.