Generation of Chemical Building Blocks from Plant Biomass by Selective Depolymerization

• US 20090004697A1
• Filed: 06/12/2008
• Published: 01/01/2009
• Est. Priority Date: 03/19/2007
• Status: Active Grant

• Current Assignee: Sued-Chemie IP GmbH & Co., KG
• Original Assignee: Sud-Chemie AG (Clariant AG)
• Inventors: Rarbach, Markus, Bruck, Thomas, Kettling, Ulrich, Koltermann, Andre
• Granted Patent: US 8,524,471 B2
• Time in Patent Office: Days
• US Class Current: 435/72
• CPC Class Codes: C12N 9/0061 Laccase (1.10.3.2)
• CPC Class Codes: C12N 9/2434 acting on beta-1,4-glucosidic bonds
• CPC Class Codes: C12P 19/02 Monosaccharides 2-ketogulonic acid C12P7/60
• CPC Class Codes: C12P 19/12 Disaccharides
• CPC Class Codes: C12P 19/14 produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase , e.g. by cellulase, hemicellulase
• CPC Class Codes: C12P 19/16 produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin non-biological hydrolysis of starch C08B30/00
• CPC Class Codes: C12P 19/18 produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
• CPC Class Codes: C12P 19/20 produced by the action of an exo-1,4 alpha-glucosidase, e.g. dextrose
• CPC Class Codes: C12P 19/22 produced by the action of a beta-amylase, e.g. maltose
• CPC Class Codes: C12P 19/24 produced by the action of an isomerase, e.g. fructose
• CPC Class Codes: C13B 35/005 using microorganisms or enzymes
• CPC Class Codes: C13K 1/02 obtained by saccharification of cellulosic materials manufacture of fodder A23K10/32
• CPC Class Codes: C13K 1/06 obtained by saccharification of starch or raw materials containing starch
• CPC Class Codes: C13K 13/00 Sugars not otherwise provided for in this class
• CPC Class Codes: C13K 13/002 Xylose

Abstract

The invention concerns a method for the enzymatic treatment of raw polymeric feedstock comprising the following steps: (a) preferably separation of soluble components from the raw polymeric feedstock, (b) treating the raw polymeric feedstock with an enzyme system in order to liberate defined soluble monomeric or oligomeric building blocks from the insoluble raw polymeric feedstock; and (c) separating the defined monomeric or oligomeric building blocks produced in step b) from the remainder of the raw polymeric feedstock. Preferably, the enzyme system used in step b) contains not more than 50%, preferably not more than 20%, more preferably not more than 10%, more preferably not more than 5%, more preferably not more than 2%, more preferably not more than 1% of other enzyme activities apart from the enzyme activity resulting in liberation of said defined monomeric or oligomeric building blocks from the raw polymeric feedstock according to step b). Further aspects of the invention concern the use of “less pure” and thus less costly enzyme systems in subsequent enzymatic treatment steps and methods for determining the optimum sequence of enzymatic treatment steps by analysis of the raw polymeric feedstock used.

Claims

1. A method for the enzymatic treatment of raw polymeric feedstock comprising:

a. treating the insoluble raw polymeric feedstock with an enzyme system in order to liberate defined soluble monomeric or oligomeric building blocks from the raw polymeric feedstock;

b. separating the defined soluble monomeric or oligomeric building blocks produced in a) from the remainder of the insoluble raw polymeric feedstock.

2. The method of claim 1, wherein the raw polymeric feedstock comprises at least 1 wt-% lignin.

3. The method of claim 2, wherein the raw polymeric feedstock comprises at least 10 wt-% lignin.

4. The method of claim 1, wherein a) and b) are repeated one or more times with different enzyme systems in order to liberate from the remainder of the raw polymeric feedstock other defined soluble monomeric or oligomeric building blocks.

5. The method of claim 1, wherein no ligninolytic enzyme treatment is performed.

6. The method of claim 1 or claim 4, wherein the content of lignin in the polymeric feedstock, calculated as wt-% of the overall composition of the polymeric feedstock is not reduced during a) and b) or their repetition.

7. The method of claim 1, wherein prior to a) soluble components are separated from the raw polymeric feedstock.

8. The method of claim 7, wherein the separation of soluble components from the raw polymeric feedstock is performed using one or more washings and/or one or more physico-chemical treatments.

9. The method of claim 1, wherein the enzyme system used in a) contains not more than 50%, of other enzyme activities apart from the enzyme activity resulting in liberation of said defined soluble monomeric or oligomeric building blocks from the insoluble raw polymeric feedstock according to a).

10. The method according to claim 4, wherein prior to repetition of a), the separation of soluble components from the processed polymeric feedstock is performed using one or more pretreatments optionally one or more washings.

11. The method of claim 1, wherein the enzymatic treatment is performed in an aqueous medium, said defined monomeric or oligomeric building blocks liberated from the raw or processed polymeric feedstock are soluble in the aqueous medium and the separation according to b) is performed by liquid/solid separation of the soluble building blocks in the aqueous medium from the remainder of the insoluble raw or processed polymeric feedstock.

12. The method of claim 1, wherein the defined monomeric or oligomeric building blocks according to a) are chosen from one of the group consisting of glucose, xylose, arabinose, galactose, mannose, amino acids and phenolic compounds.

13. The method of claim 4, wherein the feedstock is a cellulose-xylan-rich feedstock and wherein the enzyme or enzyme system used in the first enzymatic treatment of a) is a glucose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is glucose, and wherein the enzyme or enzyme system used in the second enzymatic treatment of a) is a xylose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is xylose.

14. The method according to claim 4 wherein the enzyme or enzyme system used in the first treatment of a) is selected from the group consisting of Beta-glucosidase from A. niger or from T. reesei;

Cellobiohydrolase I-II from T. reesei; and

Endo-beta-1-4-D-glucanase I-V from T. reesei, and wherein the enzyme or enzyme system used in the second treatment of a) is selected from the group consisting of Endoxylanase from A. niger or T. reesei or C. thermocellum; and

Xylosidase from A. niger or T. reesei.

15. The method of claim 4, wherein the feedstock is a cellulose-xylan-rich feedstock and wherein the enzyme or enzyme system used in the first enzymatic treatment of a) is a xylose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is xylose, and wherein the enzyme or enzyme system used in the second enzymatic treatment of a) is a glucose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is glucose.

16. The method of claim 4 wherein the enzyme or enzyme system used in the first treatment of a) is selected from the group consisting of Endoxylanase from A. niger or T. reesei or C. thermocellum;

and Xylosidase from A. niger or T. reesei, and wherein the enzyme or enzyme system used in the second treatment of a) is selected from the group consisting of Beta-glucosidase from A. niger or from T. reesei;

Cellobiohydrolase I-II from T. reesei; and

Endo-beta-1-4-D-glucanase I-V from T. reesei.

17. The method of claim 4, wherein the feedstock is an arabinan-pectin-rich feedstock and wherein the enzyme or enzyme system used in the first enzymatic treatment of a) is a arabinose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is arabinose, and wherein the enzyme or enzyme system used in the second enzymatic treatment of a) is a uronic acid-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is uronic acid.

18. The method of claim 4 wherein the enzyme or enzyme system used in the first treatment of a) is selected from the group consisting of Endoarabinase from A. niger;

and Arabinofucosidase from A. niger, and wherein the enzyme or enzyme system used in the second treatment of a) is selected from the group consisting of Pectinase, pectate lyase, and polygalactourenase from A. aculeatus, A. niger or C. japonicus.

19. The method of claim 4, wherein the feedstock is an arabinan-pectin-cellulose-rich feedstock and wherein the enzyme or enzyme system used in the first enzymatic treatment of a) is a arabinose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is arabinose, wherein the enzyme or enzyme system used in the second enzymatic treatment of a) is a glucose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is glucose, and wherein the enzyme or enzyme system used in the third enzymatic treatment of a) is a uronic acid-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is uronic acid.

20. The method of claim 4 wherein the enzyme or enzyme system used in the first treatment of a) is selected from the group consisting of Endoarabinase from A. niger;

and Arabinofucosidase from A. niger, wherein the enzyme or enzyme system used in the second treatment of a) is selected from the group consisting of Beta-glucosidase from A. niger or from T. reesei;

Cellobiohydrolase I-II from T. reesei; and

Endo-beta-1-4-D-glucanase I-V from T. reseei, and wherein the enzyme or enzyme system used in the third treatment of a) is selected from the group consisting of Pectinase, pectate lyase, and polygalactourenase from A. aculeatus, A. niger or C. japonicus.

21. The method of claim 4, wherein the feedstock is an galactan-pectin-rich feedstock and wherein the enzyme or enzyme system used in the first enzymatic treatment of a) is a galactose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is galactose, and wherein the enzyme or enzyme system used in the second enzymatic treatment of a) is a uronic acid-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is uronic acid.

22. The method of claim 4 wherein the enzyme or enzyme system used in the first treatment of a) is selected from the group consisting of Endogalactonase from A. niger or from C. thermocellum;

and beta-Galactosidase from A. niger or K. fragilis, and wherein the enzyme or enzyme system used in the second treatment of a) is selected from the group consisting of Pectinase, pectate lyase, and polygalactourenase from A. aculeatus, A. niger or C. japonicus.

23. The method of claim 4, wherein the feedstock is a mannan-xylan-rich feedstock and wherein the enzyme or enzyme system used in the first enzymatic treatment of a) is a mannose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is mannose, and wherein the enzyme or enzyme system used in the second enzymatic treatment of a) is a xylose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is xylose.

24. The method of claim 4 wherein the enzyme or enzyme system used in the first treatment of a) is selected from the group consisting of Endo-Mannanase from A. niger, B. subtilis, T. maritima or from T. reesei;

and Exo-Mannosidase from C. fimi, and wherein the enzyme or enzyme system used in the second treatment of a) is selected from the group consisting of Endoxylanase from A. niger or T. reesei or C. thermocellum; and

Xylosidase from A. niger or T. reesei.

25. The method of claim 4, wherein the feedstock is a mannan-cellulose-rich feedstock and wherein the enzyme or enzyme system used in the first enzymatic treatment of a) is a mannose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is mannose, and wherein the enzyme or enzyme system used in the second enzymatic treatment of a) is a glucose-liberating enzyme or enzyme system, and the defined soluble monomeric or oligomeric building block is glucose.

26. The method of claim 4 wherein the enzyme or enzyme system used in the first treatment of a) is selected from the group consisting of Endo-Mannanase from A. niger, B. subtilis, T. maritima or from T. reesei;

and Exo-Mannosidase from C. fimi, and wherein the enzyme or enzyme system used in the second treatment of a) is selected from the group consisting of Beta-glucosidase from A. niger or from T. reesei;

Cellobiohydrolase I-II from T. reesei; and

Endo-beta-1-4-D-glucanase I-V from T. reesei.

27. The method of claim 4, wherein the enzyme system used in a particular enzymatic treatment contains not more than 50% of contaminating enzymatic activities, which have not been employed in a previous enzymatic treatment using a different enzyme system or which can cause liberation of other defined monomeric or oligomeric building blocks, which have not been liberated in previous enzymatic treatments, or which can only cause liberation of products from polymeric substrates that are initially essentially absent from the polymeric feedstock.

28. The method of claim 4, wherein the raw polymeric feedstock comprises cellulose and hemicellulose as polymeric substrates and the enzyme system used in a particular enzymatic treatment has cellulase activity, and optionally beta-glycosidase, glucohydrolase and/or alpha- or beta-amylase activity, but is essentially free of hemicellulase activity.

29. The method of claim 4, wherein the enzyme system used in a particular enzymatic treatment contains as contaminating enzymatic activities one or more of such enzymatic activities, which have been employed in a previous enzymatic treatment using a different enzyme system or which can only cause liberation of other monomeric or oligomeric building blocks from polymeric substrates that are initially essentially absent in the raw polymeric feedstock.

30. The method of claim 4, wherein the enzyme system used in a particular enzymatic treatment has a first enzymatic activity and contains at least one additional enzymatic activity, which leads to the same defined monomeric or oligomeric building block from the raw or processed polymeric feedstock as the first enzymatic activity of the enzyme system.

31. The method of claim 1 or claim 4, wherein the insoluble raw or processed polymeric feedstock is subjected to a selective or unselective physico-chemical treatment prior to a) or prior to repetition of a).

32. The method according to claim 31, wherein the physico-chemical treatment comprises a treatment with aqueous solvents, organic solvents, or any combination or mixtures of these with ethanol or glycerol.

33. The method of claim 1 or claim 4, wherein the enzyme system giving the highest purity of the defined monomeric or oligomeric building blocks after treatment of the raw polymeric feedstock with the enzyme system and separation of the defined monomeric or oligomeric building blocks from the remainder of the raw polymeric feedstock is chosen for the first enzymatic treatment according to a).

34. The method according to claim 4, wherein the second enzyme system chosen for the second enzymatic treatment according to a) is one which gives the highest purity of the defined monomeric or oligomeric building blocks after treatment of the processed polymeric feedstock obtained with the enzyme system and separation of the defined monomeric or oligomeric building blocks from the raw polymeric feedstock.

35. The method according to claim 34, wherein any subsequent enzyme treatments are performed in the order of decreasing purity of the defined monomeric or oligomeric building blocks obtained after treatment of the processed polymeric feedstock obtained from the previous enzyme treatment with the respective enzyme system and separation of the defined monomeric or oligomeric building blocks from the processed polymeric feedstock.

36. The method of claim 4, wherein the enzyme systems to be employed and their sequence of use is determined by first analyzing the raw polymeric feedstock.

37. The method according to claim 36, wherein, in order to determine the enzyme systems to be used and their sequence, the insoluble raw polymeric feedstock is

i. first treated in separate enzymatic treatments with each of a plurality of enzyme systems;

ii. for each enzymatic treatment the defined monomeric or oligomeric building blocks liberated from the raw polymeric feedstock are analyzed for purity of the defined monomeric or oligomeric building blocks after separation of the defined monomeric or oligomeric building blocks from the raw polymeric feedstock;

iii. the enzyme system giving the highest purity is chosen for the first enzymatic treatment according to a);

iv. optionally the sequence of i. to iii. is repeated with the remainder of the raw or processed polymeric feedstock in order to determine the enzyme system to be used in the subsequent enzymatic treatment.

38. The method according to claim 36, wherein, in order to determine the enzyme systems to be used and their sequence, the insoluble raw polymeric feedstock is

i. first treated in separate enzymatic treatments with each of a plurality of enzyme systems to determine the monomeric or oligomeric building blocks contributing the largest mass ratio in the raw polymeric feedstock;

ii. the defined monomeric or oligomeric building blocks contributing the largest mass ratio in the raw polymeric feedstock are analyzed for purity after separation from the raw polymeric feedstock;

iii. if the purity as determined is such that more than 75 wt. of the total solids content consists of the defined monomeric or oligomeric building blocks, the respective enzyme system is chosen for the first enzymatic treatment according to a);

iv. if the purity determined according to ii. is lower than required according to iii., the defined monomeric or oligomeric building blocks contributing the next largest mass ratio in the raw polymeric feedstock are analyzed for purity after separation from the raw polymeric feedstock accordingly, until a purity satisfies the requirement according to iii. and the respective enzyme system is chosen for the first enzymatic treatment according to a);

v. optionally the sequence of i. to iv. is repeated with the remainder of the raw or processed polymeric feedstock in order to determine the enzyme system to be used in the subsequent enzymatic treatment.

39. The method according to claim 36, wherein, in order to determine the enzyme systems to be used and their sequence, the raw polymeric feedstock is

i. first treated in separate enzymatic treatments with each of a plurality of enzyme systems to determine the enzymatic treatment that leads to the highest yield of monomeric or oligomeric building blocks contained in the raw polymeric feedstock;

ii. select those enzymatic treatments that yield a defined monomeric or oligomeric product with a purity of more than 75. wt. of the total solids;

iii. determine among the remaining enzymatic treatments the treatment with the highest yield of monomeric or oligomeric product;

iv. optionally the sequence of i. to iii. is repeated with the remainder of the raw or processed polymeric feedstock in order to determine the enzyme system to be used in the subsequent enzymatic treatment.

41. The method according to any one of claims 36 to 39 wherein the enzyme systems comprise enzymes liberating soluble monomeric or oligomeric saccharide building blocks from the raw polymeric feedstock.

40. (canceled)