US2016273012A1PendingUtilityA1

Glucosyltransferase enzymes for production of glucan polymers

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Assignee: DU PONTPriority: Sep 25, 2012Filed: Mar 25, 2016Published: Sep 22, 2016
Est. expirySep 25, 2032(~6.2 yrs left)· nominal 20-yr term from priority
Inventors:Mark S. PayneYefim BrunHongxian HeThomas Scholz
C12Y 204/01267C12P 19/04C12P 19/18C12N 9/1051C12Y 204/01005Y02P20/52C08B 37/0009C12N 9/1048
68
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Claims

Abstract

Reaction solutions are disclosed herein comprising water, sucrose and a glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan. The glucosyltransferase enzyme can synthesize insoluble glucan polymer having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. Further disclosed are methods of using such glucosyltransferase enzymes to produce insoluble poly alpha-1,3-glucan.

Claims

exact text as granted — not AI-modified
1 . A reaction solution comprising water, sucrose and a non-native glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         2 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. 
     
     
         3 . The reaction solution of  claim 2 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having at least 95% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. 
     
     
         4 . The reaction solution of  claim 3 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having 100% alpha-1,3 glycosidic linkages. 
     
     
         5 . The reaction solution of  claim 1 , further comprising a primer. 
     
     
         6 . The reaction solution of  claim 5 , wherein the primer is dextran. 
     
     
         7 . The reaction solution of  claim 5 , wherein the primer is hydrolyzed glucan. 
     
     
         8 . A method for producing poly alpha-1,3-glucan comprising:
 a) contacting at least water, sucrose, and a non-native glucosyltransferase enzyme that synthesizes poly alpha-1,3-glucan, wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 90% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34;   whereby poly alpha-1,3-glucan is produced; and   b) optionally, isolating the poly alpha-1,3-glucan produced in step (a).   
     
     
         9 . The method of  claim 8 , wherein said glucosyltransferase enzyme synthesizes poly alpha-1,3-glucan having at least 50% alpha-1,3 glycosidic linkages and a number average degree of polymerization of at least 100. 
     
     
         10 - 14 . (canceled) 
     
     
         15 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 91° A identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         16 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 92% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         17 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 93% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         18 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 94% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         19 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 95% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         20 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 96% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         21 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 97% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         22 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 98% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         23 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme comprises an amino acid sequence that is at least 99% identical to SEQ ID NO:4, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:20, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, or SEQ ID NO:34. 
     
     
         24 . The reaction solution of  claim 1 , wherein said glucosyltransferase enzyme does not comprise amino acid residues 2-1477 or 138-1477 of SEQ ID NO:8. 
     
     
         25 . The reaction solution of  claim 1 , wherein a heterologous amino acid sequence of 1-300 residues is at the N-terminus and/or C-terminus of said glucosyltransferase enzyme.

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