Compositions comprising a polypeptide having cellulolytic enhancing activity and a heterocyclic compound and uses thereof

Patent metadata

Field	Value
Publication number	US-10041101-B2
Application number	US-201615193434-A
Country	US
Kind code	B2
Filing date	Jun 27, 2016
Priority date	Aug 12, 2010
Publication date	Aug 7, 2018
Grant date	Aug 7, 2018

Abstract

The present invention relates to compositions comprising: a polypeptide having cellulolytic enhancing activity and a heterocyclic compound. The present invention also relates to methods of using the compositions.

First claim

What is claimed is:

1. A method for producing a fermentation product, comprising:
(a) saccharifying a cellulosic material with an enzyme composition comprising a glycoside hydrolase family 61 (GH61) polypeptide having cellulolytic enhancing activity and a heterocyclic compound, wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the heterocyclic compound enhances hydrolysis of the cellulosic material by the enzyme composition compared to the GH61 polypeptide having cellulolytic enhancing activity alone or the heterocyclic compound alone with a ratio greater than 1 according to the following formula:
GH 61 effect=% conversion (+GH61+heterocyclic compound) /% conversion (no GH61+heterocyclic compound)

wherein the heterocyclic compound is a compound of formula (I) or (II):

wherein each bond indicated with a dashed line is single or double;
R 1 , R 2 , R 3 , R 4 , and R 5 are independently hydrogen, halogen, ═O, —OH, —OR 8 , —CN, —NO 2 , —N(R 9 )(R 10 ), —C(O)R 20 , —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 21 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl;
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 , R 20 , and R 21 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; and
wherein each pair of R 1 and R 2 , R 2 and R 3 , R 3 and R 4 , and R 4 and R 5 may combine to form an optionally substituted fused ring;
or a salt or solvate thereof;
(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.

2. The method of claim 1 , wherein the cellulosic material is pretreated.

3. The method of claim 1 , wherein steps (a) and (b) are performed simultaneously in a simultaneous saccharification and fermentation.

4. The method of claim 1 , 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.

5. The method 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 method 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. The method of claim 1 , wherein the heterocyclic compound is selected from the group consisting of: (I-1): (1,2-dihydroxyethyl)-3,4-dihydroxyfuran-2(5H)-one; (I-2): 4-hydroxy-5-methyl-3-furanone; (I-3): 5-hydroxy-2(5H)-furanone; (I-4): [1,2-dihydroxyethyl]furan-2,3,4(5H)-trione; (I-5): α-hydroxy-γ-butyrolactone; (I-6): ribonic γ-lactone; (I-7): glucuronic acid γ-lactone; (I-8): dihydrobenzofuran; (I-9): 5-(hydroxymethyl)furfural; (I-10): furoin; (I-11): 2(5H)-furanone; (II-1): gluconic acid δ-lactone; (II-2): 4-hydroxycoumarin; (II-3): 5,6-dihydro-2H-pyran-2-one; (II-4): 5,6-dihydro-4-hydroxy-6-methyl-2H-pyran-2-one; (II-5): 1,5-anhydro-2-deoxy-arabino-hex-1-enitol; and (II-6): 3-deoxy-erythro-hexosulose; 3-hydroxy-5-methylisoxazole; or a salt or solvate thereof.

8. 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.

9. The method of claim 1 , wherein the heterocyclic compound is a compound of formula (I-A), (I-B), or (I-C):

wherein R 1 , R 2 , R 3 , and R 4 are as defined in the preceding claims; or a salt or solvate thereof.

10. The method of claim 1 , wherein the heterocyclic compound is a compound of formula (I-D), (I-E), (I-F), or (I-G):

wherein R 1 , R 2 , R 3 , and R 4 are as defined in the preceding claims; or a salt or solvate thereof.

11. The method of claim 1 , wherein the heterocyclic compound is a compound of formula (I-A), (I-B), or (I-C):

wherein R 1 , R 2 , R 3 , R 4 , and R 5 are as defined in the preceding claims; or a salt or solvate thereof.

12. The method of claim 1 , wherein an effective amount of the heterocyclic compound to cellulosic material as a molar ratio to glucosyl units of cellulose is about 10 −6 to about 10.

13. The method of claim 1 , wherein an effective amount of the heterocyclic compound to cellulose is about 10 −6 to about 10 g per g of cellulose.

14. A method of fermenting a saccharified cellulosic material, comprising: fermenting the saccharified cellulosic material with one or more fermenting microorganisms, wherein the saccharified cellulosic material is obtained by saccharifying a cellulosic material with an enzyme composition comprising a GH61 polypeptide having cellulolytic enhancing activity and a heterocyclic compound, wherein the combination of the GH61 polypeptide having cellulolytic enhancing activity and the heterocyclic compound enhances hydrolysis of the cellulosic material by the enzyme composition compared to the GH61 polypeptide having cellulolytic enhancing activity alone or the heterocyclic compound alone with a ratio greater than 1 according to the following formula:
GH 61 effect=% conversion (+GH61+heterocyclic compound) /% conversion (no GH61+heterocyclic compound)
wherein the heterocyclic compound is a compound of formula (I) or (II):

wherein each bond indicated with a dashed line is single or double;
R 1 , R 2 , R 3 , R 4 , and R 5 are independently hydrogen, halogen, ═O, —OH, —OR 8 , —CN, —NO 2 , —N(R 9 )(R 10 ), —C(O)R 20 , —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 21 , or an optionally substituted moiety selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl-alkyl, heterocycloalkyl, heterocycloalkyl-alkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl;
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 , R 20 , and R 21 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-alky

Assignees

• Novozymes Inc

Inventors

• Quinlan Jason
• Xu Feng
• Sweeney Matthew

Classifications

• C12N9/96

  Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates · CPC title

• C12Y302/01004

  Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase · CPC title

• C12P7/28

  Acetone-containing products · CPC title

• C12P2203/00

  Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source · CPC title

• C12P2201/00

  Pretreatment of cellulosic or lignocellulosic material for subsequent enzymatic treatment or hydrolysis · CPC title