Glucanases, nucleic acids encoding them and methods for making and using them

• US 10,329,549 B2
• Filed: 03/28/2014
• Issued: 06/25/2019
• Est. Priority Date: 08/04/2006
• Status: Active Grant

• Current Assignee: BP Corporation North America Incorporated (BP PLC)
• Original Assignee: BP Corporation North America Incorporated (BP PLC)
• Inventors: Steer, Brian, Healey, Shaun, Esteghlalian, Alireza, Miles, Stacy Marie, Barrett, Kenneth, Quadt, Rene
• Primary Examiner(s): Tsay, Marsha
• Application Number:
  US14/229,633
• Publication Number:
  US 20140295523A1
• Time in Patent Office: 1,915 Days
• CPC Class Codes: A01H 6/14 Asteraceae or Compositae, e.g. safflower, sunflower, artichoke or lettuce
• CPC Class Codes: A01H 6/46 Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
• CPC Class Codes: A23K 20/147 Polymeric derivatives, e.g. peptides or proteins
• CPC Class Codes: A23L 33/18 Peptides; Protein hydrolysates
• CPC Class Codes: A61P 1/14 Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C09K 2208/24 Bacteria or enzyme containing gel breakers
• CPC Class Codes: C09K 8/035 Organic additives
• CPC Class Codes: C09K 8/52 Compositions for preventing, limiting or eliminating depositions, e.g. for cleaning
• CPC Class Codes: C09K 8/68 containing organic compounds
• CPC Class Codes: C09K 8/74 combined with additives added for specific purposes
• CPC Class Codes: C09K 8/805 Coated proppants
• CPC Class Codes: C09K 8/90 of natural origin, e.g. polysaccharides, cellulose
• CPC Class Codes: C12N 9/2405 Glucanases
• CPC Class Codes: C12N 9/2437 Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
• CPC Class Codes: C12P 19/02 Monosaccharides 2-ketogulonic acid C12P7/60
• 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: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: C12Y 302/01006 : Endo-1,3(4)-beta-glucanase (3.2.1.6)
• CPC Class Codes: C12Y 302/01008 : Endo-1,4-beta-xylanase (3.2.1.8)
• CPC Class Codes: C12Y 302/01039 : Glucan endo-1,3-beta-D-glucosidase (3.2.1.39)
• CPC Class Codes: E21B 43/26 : by forming crevices or fractures
• CPC Class Codes: G16B 30/00 : ICT specially adapted for sequence analysis involving nucleotides or amino acids
• CPC Class Codes: G16B 30/10 : Sequence alignment; Homology search
• CPC Class Codes: G16B 30/20 : Sequence assembly
• CPC Class Codes: Y02E 50/10 : Biofuels, e.g. bio-diesel

Abstract

The invention relates to polypeptides having glucanase, e.g., endoglucanase, mannanase, xylanase activity or a combination of these activities, and polynucleotides encoding them. In one aspect, the glucanase activity is an endoglucanase activity (e.g., endo-1,4-beta-D-glucan 4-glucano hydrolase activity) and comprises hydrolysis of 1,4-beta-D-glycosidic linkages in cellulose, cellulose derivatives (e.g., carboxy methyl cellulose and hydroxy ethyl cellulose) lichenin, beta-1,4 bonds in mixed beta-1,3 glucans, such as cereal beta-D-glucans or xyloglucans and other plant material containing cellulosic parts. In addition, methods of designing new enzymes and methods of use thereof are also provided. In alternative aspects, the new glucanases e.g., endoglucanases, mannanases, xylanases have increased activity and stability, including thermotolerance or thermostability, at increased or decreased pHs and temperatures.

Claims

1. A synthetic or recombinant polypeptide with endoglucanase activity comprising:

(a) the amino acid sequence as set forth in SEQ ID NO;

9;

(b) the synthetic or recombinant polypeptide encoded by the nucleic acid sequence as set forth in SEQ ID NO;

8;

or

(c) the polypeptide of (a) or the polypeptide encoded by (b) but lacking a signal sequence or a carbohydrate binding module;

wherein, the polypeptide further comprises a heterologous sequence.

2. The polypeptide of claim 1, wherein the endoglucanase activity is thermostable or thermotolerant.

3. A food or feed composition and/or food additive comprising the polypeptide of claim 1.

4. A method for preparing a food or feed composition and/or food or feed additive, comprising mixing the polypeptide of claim 1 with one or more food or feed and/or food or feed additive ingredients.

5. A detergent composition comprising the polypeptide of claim 1.

6. The detergent composition of claim 5 further comprising one or more additional polypeptides.

7. The detergent composition of claim 5 or 6 additionally comprising a surfactant.

8. A fracturing fluid composition comprising:

(a) an aqueous fluid,

(b) a hydratable polymer; and

(c) the polypeptide of claim 1.

9. The fracturing fluid of claim 8 wherein the composition further comprises a crosslinking agent capable of cross-linking the hydratable polymer.

10. The fracturing fluid of claim 8 or 9, wherein the hydratable polymer is selected from the group consisting of guar, guar derivatives, cellulose derivatives, water soluble biopolymers, or combinations thereof.

11. The fracturing fluid of claim 8, 9, or 10, further comprising one or more additional polypeptides.

12. A method of fracturing a subterranean formation that surrounds a well bore, the method comprising the steps of:

(a) combining an aqueous fluid, a hydratable polymer, a crosslinking agent, and an enzyme breaker, comprising the polypeptide of claim 1;

(b) injecting the crosslinked polymer fluid into the well bore and into contact with the formation under sufficient pressure to fracture the surrounding subterranean formation; and

(c) allowing the enzyme breaker to degrade the crosslinked polymer gel so that it can be removed from the subterranean formation, the enzyme breaker being catalytically active and temperature stable in a temperature range of about 60°

F. to about 225°

F. and in a pH range of about 7 to about 12.

13. A method of modifying a composition comprising lignocellulose, a cellulose, a hemicellulose, a lignin, a xylan, a glucan, or a mannan comprising contacting the composition with the polypeptide of claim 1.

16. A cellulose, wood, wood pulp, wood product, paper, paper pulp, paper product, pharmaceutical composition, dietary supplement, fuel, dairy product, textile, fabric , processed waste, disinfectant, biodefense, bio-detoxifying agent comprising the polypeptide of claim 1.

17. A method of hydrolyzing or disrupting a cellulose containing composition comprising contacting the composition with the polypeptide of claim 1, thereby hydrolyzing or disrupting the composition.

14. A polypeptide having endoglucanase activity comprising SEQ ID NO:

9.

15. A polypeptide having endoglucanase activity encoded by SEQ ID NO:

8.

18. The polypeptide of claim 15, wherein the endoglucanase activity is thermostable.

19. A composition comprising a mixture of SEQ ID NO:

9 or a polypeptide having at least 98% or more sequence identity to SEQ ID NO;

9 and at least one or more additional polypeptides.

20. A method of hydrolyzing or disrupting a cellulose containing composition comprising contacting the cellulose containing composition with a composition of claim 19, thereby hydrolyzing or disrupting the cellulose containing composition.