Compositions comprising a polypeptide having cellulolytic enhancing activity and a bicyclic compound and uses thereof
The present invention relates to compositions comprising: a polypeptide having cellulolytic enhancing activity and a bicyclic compound. The present invention also relates to methods of using the compositions.
1. A method for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with an enzyme composition in the presence of a GH61 polypeptide having cellulolytic enhancing activity and a bicyclic compound;
(b) fermenting the saccharified cellulosic material with one or more fermenting microorganisms to produce the fermentation product; and
(c) recovering the fermentation product from the fermentation,
wherein the bicyclic compound is of formula (I) or (II):
GH
61
effect
=
%
conversion
(
+
GH
61
)
%
conversion
(
no
GH
61
)
.
wherein the bond indicated with a dashed line can be single or double;
R 1 and R 2 are independently hydrogen, —C(O)R 5 , —C(O)OR 6 , —C(O)NHR 7 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl;
R 3 , R 4a , R 4b , and R 4c are independently hydrogen, halogen, —OH, —OR 8 , —CN, —NO 2 , —N(R 9 )(R 10 ), —C(O)R 5 , —C(O)OR 6 , —C(O)NHR 7 , —OC(O)R 11 , —NHC(O) R 12 , —OC(O)OR 13 , —NHC(O)OR 14 , —OC(O)NHR 15 , —NHC(O)NHR 16 , —SO 2 R 17 , —SO 2 N(R 18 )(R 19 ), —SR 20 , 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 4a , R 4b , and R 4c may together form an optionally substituted fused ring;
R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 18 , R 19 , and R 20 are independently hydrogen, or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl; and
R 17 is 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;
wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the bicyclic compound enhances hydrolysis of a cellulosic material by the enzyme composition according to the following formula with a ratio greater than 1:
GH
61
effect
=
%
conversion
(
+
GH
61
)
%
conversion
(
no
GH
61
)
.
2. The method of claim 1 , wherein the bicyclic compound is of the formula (I-A) or (II-A):
or a salt or solvate thereof.
3. The method of claim 1 , wherein the bicyclic compound is of the formula (I-B) or (II-B):
or a salt or solvate thereof.
4. The method of claim 1 , wherein the bicyclic compound is of the formula (I-C), (I-D), (I-E), or (I-F):
or a salt or solvate thereof.
5. The method of claim 1 , wherein the bicyclic compound is selected from the group consisting of: (I-1): epicatechin; (I-2): quercetin; (I-3): myricetin; (I-4): taxifolin; (I-5): kaempferol; (I-6): morin; (I-7): acacetin; (I-8): naringenin; (I-9): isorhamnetin; (I-10): apigenin; (II-1): cyanidin; (II-2): cyanin; (II-3): turomanin; and (II-4): keracyanin; or a salt or solvate thereof.
6. The method of claim 1 , wherein an effective amount of the bicyclic compound is about 10 −6 to about 10 g per g of cellulose.
7. The method of claim 1 , wherein the enzyme composition 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.
8. The method of claim 7 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.
9. The method of claim 7 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetylxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.
10. The method of claim 1 , wherein the cellulosic material is pretreated.
11. The method of claim 1 , wherein the fermentation product is an alcohol, an alkane, a cycloalkane, an alkene, an amino acid, a gas, isoprene, a ketone, an organic acid, or polyketide.
12. A method of fermenting a cellulosic material, comprising: fermenting the cellulosic material with one or more fermenting microorganisms, wherein the cellulosic material is saccharified with an enzyme composition in the presence of a polypeptide having cellulolytic enhancing activity and a bicyclic compound,
wherein the bicyclic compound is of formula (I) or (II):
wherein the bond indicated with a dashed line can be single or double;
R 1 and R 2 are independently hydrogen, —C(O)R 5 , —C(O)OR 6 , —C(O)NHR 7 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl;
R 3 , R 4a , R 4b , and R 4c are independently hydrogen, halogen, —OH, —OR 8 , —CN, —NO 2 , —N(R 9 )(R 10 ), —C(O)R 5 , —C(O)OR 6 , —C(O)NHR 7 , —OC(O)R 11 , —NHC(O) R 12 , —OC(O)OR 13 , NHC(O)OR 14 , —OC(O)NHR 15 , —NHC(O)NHR 16 , —SO 2 R 17 , —SO 2 N(R 18 )(R 19 ), —SR 20 , 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 4a , R 4b , and R 4c may together form an optionally substituted fused ring;
R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 18 , R 19 , and R 20 are independently hydrogen, or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl; and
R 17 is 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;
wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the bicyclic compound enhances hydrolysis of a cellulosic material by the enzyme composition according to the following formula with a ratio greater than 1:
GH
61
effect
=
%
conversion
(
+
GH
61
)
%
conversion
(
no
GH
61
)
.
13. The method of claim 12 , wherein the bicyclic compound is of the formula (I-A) or (II-A):
or a salt or solvate thereof.
14. The method of claim 12 , wherein the bicyclic compound is of the formula (I-B) or (II-B):
or a salt or solvate thereof.
15. The method of claim 12 , wherein the bicyclic compound is of the formula (I-C), (I-D), (I-E), or (I-F):
or a salt or solvate thereof.
16. The method of claim 12 , wherein the bicyclic compound is selected from the group consisting of: (I-1): epicatechin; (I-2): quercetin; (I-3): myricetin; (I-4): taxifolin; (I-5): kaempferol; (I-6): morin; (I-7): acacetin; (I-8): naringenin; (I-9): isorhamnetin; (I-10): apigenin; (II-1): cyanidin; (II-2): cyanin; (II-3): turomanin; and (II-4): keracyanin; or a salt or solvate thereof.
17. The method of claim 12 , wherein an effective amount of the bicyclic compound is about 10 −6 to about 10 g per g of cellulose.
18. The method of claim 12 , wherein the enzyme composition 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.
19. The method of claim 18 , wherein the cellulase is one or more enzymes selected from the group consisting of an endoglucanase, a cellobiohydrolase, and a beta-glucosidase.
20. The method of claim 18 , wherein the hemicellulase is one or more enzymes selected from the group consisting of a xylanase, an acetylxylan esterase, a feruloyl esterase, an arabinofuranosidase, a xylosidase, and a glucuronidase.
21. The method of claim 12 , wherein the cellulosic material is pretreated.
22. The method of claim 12 , wherein the fermenting of the cellulosic material produces a fermentation product.
23. The method of claim 22 , further comprising recovering the fermentation product from the fermentation.
24. The method of claim 22 , wherein the fermentation product is an alcohol, an alkane, a cycloalkane, an alkene, an amino acid, a gas, isoprene, a ketone, an organic acid, or polyketide.