Compositions comprising a polypeptide having cellulolytic enhancing activity and a nitrogen-containing compound and uses thereof
The present invention relates to compositions comprising: a polypeptide having cellulolytic enhancing activity and a nitrogen-containing compound. The present invention also relates to methods of using the compositions.
1. An enzyme composition for degrading a cellulosic material to fermentable sugars, comprising: (a) a GH61 polypeptide having cellulolytic enhancing activity and (b) a nitrogen-containing compound, wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the nitrogen-containing compound enhances hydrolysis of a cellulosic material by a cellulolytic enzyme compared to the GH61 polypeptide alone or the nitrogen-containing compound alone, and wherein the nitrogen-containing compound is a compound of formula (I) or (II-A):
wherein R 1 and R 2 are each independently hydrogen, or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteraralkyl; or wherein R 1 and R 2 together with the attached carbon form an optionally substituted aromatic or non-aromatic ring;
wherein R 3 and R 4 are each independently hydrogen, an oxygen radical, —C(O)R 5 , —C(O)NHR 6 , —C(O)OR 7 , —SO 2 R 8 , —OR 9 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteraralkyl; or wherein R 3 and R 4 , R 3 and R 1 , or R 4 and R 2 together with the attached nitrogen form an optionally substituted aromatic or non-aromatic ring;
wherein R 9A , R 9B , and R 9C are independently hydrogen, halogen, —OH, —OR 10 , —CN, —NO 2 , —N(R 11 )(R 12 ), —C(O)R 13 , —C(O)OR 14 , —C(O)NHR 15 , —OC(O)R 16 , —NHC(O)R 17 , —OC(O)OR 18 , —NHC(O)OR 19 , —OC(O)NHR 20 , —NHC(O)NHR 21 , —SO 2 R 22 , —SO 2 N(R 23 )(R 24 ), —SR 25 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl; and wherein two of R 9A , R 9B , and R 9C may together form an optionally substituted fused ring; and
wherein R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 21 , R 22 , R 23 , R 24 , and R 25 are independently hydrogen, or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl;
or a salt or solvate thereof;
or (II-3): 2,2,6,6-tetramethyl-1-piperidinyloxy or (II-6): maleamic acid; or a salt or solvate thereof.
2. The enzyme composition of claim 1 , wherein the nitrogen-containing compound comprises an amine, imine, hydroxylamine, or nitroxide moiety.
3. The enzyme composition of claim 1 , wherein the nitrogen-containing compound is selected from the group consisting of: (I-1): acetone oxime; (I-2): violuric acid; (I-3): pyridine-2-aldoxime; (II-1): 2-aminophenol; and (II-2): 1,2-benzenediamine; or a salt or solvate thereof.
4. The enzyme composition of claim 1 , which further comprises (c) one or more enzymes selected from the group consisting of a cellulase, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.
5. The enzyme composition of claim 4 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.
6. The enzyme composition of claim 4 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetyxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.
7. A method for degrading a cellulosic material to fermentable sugars, comprising: treating the cellulosic material with an enzyme composition comprising: (a) a GH61 polypeptide having cellulolytic enhancing activity and (b) a nitrogen-containing compound, wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the nitrogen-containing compound enhances hydrolysis of a cellulosic material by a cellulolytic enzyme compared to the GH61 polypeptide alone or the nitrogen-containing compound alone, and wherein the nitrogen-containing compound is a compound of formula (I) or (II-A):
wherein R 1 and R 2 are each independently hydrogen, or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteraralkyl; or wherein R 1 and R 2 together with the attached carbon form an optionally substituted aromatic or non-aromatic ring;
wherein R 3 and R 4 are each independently hydrogen, an oxygen radical, —C(O)R 5 , —C(O)NHR 6 , —C(O)OR 7 , —SO 2 R 8 , —OR 9 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteraralkyl; or wherein R 3 and R 4 , R 3 and R 1 , or R 4 and R 2 together with the attached nitrogen form an optionally substituted aromatic or non-aromatic ring;
wherein R 9A , R 9B , and R 9C are independently hydrogen, halogen, —OH, —OR 10 , —CN, —NO 2 , —N(R 11 )(R 12 ), —C(O)R 13 , —C(O)OR 14 , —C(O)NHR 15 , —OC(O)R 16 , —NHC(O)R 17 , —OC(O)OR 18 , —NHC(O)OR 19 , —OC(O)NHR 20 , —NHC(O)NHR 21 , —SO 2 R 22 , —SO 2 N(R 23 )(R 24 ), —SR 25 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl; and wherein two of R 9A , R 9B , and R 9C may together form an optionally substituted fused ring; and
wherein R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R 20 , R 21 , R 22 , R 23 , R 24 , and R 25 are independently hydrogen, or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl;
or a salt or solvate thereof;
or (II-3): 2,2,6,6-tetramethyl-1-piperidinyloxy or (II-6): maleamic acid; or a salt or solvate thereof; and
recovering the degraded cellulosic material;
wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the nitrogen-containing compound enhances hydrolysis of the cellulosic material by the enzyme composition according to the following formula with a ratio greater than 1:
GH
61
effect
=
%
conversion
(
+
GH6
1
+
nitrogen-containing
compound
)
%
conversion
(
noGH6
1
+
nitrogen-containing
compound
)
.
8. The method of claim 7 , wherein the enzyme composition further comprises one or more enzymes selected from the group consisting of a cellulase, a hemicellulase, an esterase, an expansin, a laccase, a ligninolytic enzyme, a pectinase, a peroxidase, a protease, and a swollenin.
9. The method of claim 8 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.
10. The method of claim 8 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetyxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.
11. The method of claim 7 , wherein the nitrogen-containing compound comprises an amine, imine, hydroxylamine, or nitroxide moiety.
12. The method of claim 7 , wherein the nitrogen-containing compound is selected from the group consisting of: (I-1): acetone oxime; (I-2): violuric acid; (I-3): pyridine-2-aldoxime; (II-1): 2-aminophenol; and (II-2): 1,2-benzenediamine; or a salt or solvate thereof.
13. The method of claim 7 , wherein the cellulosic material is pretreated.
14. The method of claim 7 , wherein the degraded cellulosic material is a fermentable sugar.
15. The method of claim 14 , wherein the fermentable sugar is selected from the group consisting of glucose, xylose, mannose, galactose, and arabinose.