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Thermoacidophilic derived enzymes for biomass conversion to fuels and chemicals

Improving the economics of biomass preprocessing

Idaho National Laboratory

Contact INL About This Technology

Publications:

PDF Document PublicationXylanase Fact Sheet (246 KB)

Xylanase activity
Xylanase activity

Technology Marketing Summary

INL’s thermoacidophilic enzyme suite has potential to improve the cost and efficiency of multiple industrial bio-processes. Commercial enzymes available today are generally active between 25°C and 50°C at a pH of about 5. These conditions are not aligned with reaction conditions in many industrial processes. In order to make use of enzyme catalysts in such systems, excess enzymes dosages are required or additional unit operations are added to cool and adjust the pH of reactants, each of which add substantial cost. One example occurs in the animal feed industry where several of types of enzymes are added to make the feed more digestible. After enzymes are added, the feed is pelleted in a high temperature process (100-120°C), destroying a portion of the enzyme.  In order to compensate, excess enzyme must be added. Similar conditions exist in other industrial processes that require enzymes such as detergent additives, wine or beer processing, pulp and paper bleaching, food and juice processing, baking, dairy processing, and custom prebiotic syntheses. Using thermo- and acid-tolerant enzymes has the potential to significantly improve efficiency of such industrial processes.

INL’s Alicyclobacillus derived enzymes are active at temperatures up to 70°C and pHs from 2–8.  Activity at 70°C, pH 6 can be up to 8 times greater than other commercially available enzymes. Using Alicyclobacillus enzymes reduces cost by decreasing enzyme loading since more enzymes remain viable in high temperature, low pH conditions. Further characterization and enzyme cocktail optimization is necessary to understand performance in specific applications. 

Description

This INL technology includes a suite of lignocellulose degrading enzymes which operate at higher temperatures and/or lower pHs than current commercial lignocellulose enzymes. The thermoacidophile, Alicyclobacillus acidocaldarius, produces these enzymes which release sugars from biomass by breaking linkages between hemicellulose and lignin to free hemicellulose from the biomass matrix, cleaving sidechains in hemicellulose that block its breakdown, hydrolyzing cellulose, and hemicellulose to release cellobiose and xylobiose and cleaving cellobiose and xylobiose to generate glucose and xylose. In biorefineries, these sugars can then be used to produce fuels and chemicals through fermentation.

Additionally, the enzymes produced by Alicyclobacillus can be utilized as standalone products individually or in combination for other industrial applications such as detergent additives, animal feed additives, wine or beer processing, pulp and paper bleaching, food and juice processing, baking, dairy processing, and custom prebiotic syntheses. The Alicyclobacillus enzymes are more robust than current commercially available enzymes and operate at industrially relevant temperatures and/or pHs. All of the enzymes can be produced and purified in at least milligram quantities and in many cases gram to kilogram quantities and can be made available for testing.

Benefits
  • Reduced operating cost by decreasing required enzyme loading in industrial processes.
  • Allows for the higher temperature, lower pH process conditions to improve reactor efficiency.
  • Eliminates the need for costly unit operations to reduce reactant temperature and increase pH to be compatible with enzyme catalysts.

 

Applications and Industries
  • Biofuel / biochemical production
  • Food processing
  • Pulp & Paper
  • Detergents
More Information

Additional development will be required to demonstrate and scale this technology for commercial applications. BEA may be in a position to support additional research and development of this technology under a mutually acceptable Cooperative Research & Development Agreement (CRADA), Strategic Partnership Projects (SPP) agreement, or a similar, all of which are subject to approval by DOE.

Patents and Patent Applications
ID Number
Title and Abstract
Primary Lab
Date
Patent 7,727,755
Patent
7,727,755
Enzyme and methodology for the treatment of a biomass
An enzyme isolated from an extremophilic microbe, and a method for utilizing same is described, and wherein the enzyme displays optimum enzymatic activity at a temperature of greater than about 80.degree. C., and a pH of less than about 2, and further may be useful in methodology including pretreatment of a biomass so as to facilitate the production of an end product.
Idaho National Laboratory 06/01/2010
Issued
Patent 7,923,234
Patent
7,923,234
Thermal and acid tolerant beta-xylosidases, genes encoding, related organisms, and methods
Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius and variations thereof are provided. Further provided are methods of at least partially degrading xylotriose and/or xylobiose using isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius and variations thereof.
Idaho National Laboratory 04/12/2011
Issued
Patent 9,862,981
Patent
9,862,981
Thermophilic acetylxylan esterase genes and enzymes from alicyclobacillus acidocaldarius and related organisms and methods
A genetically modified organism comprising at least one nucleic acid sequence and/or at least one recombinant nucleic acid isolated from Alicyclobacillus acidocaldarius and encoding a polypeptide involved in at least partially degrading, cleaving, transporting, metabolizing, or removing polysaccharide, lignocellulose, hemicellulose, lignin, chitin, heteroxylan, and/or xylan-decorating group; and at least one nucleic acid sequence and/or at least one recombinant nucleic acid encoding a polypeptide involved in fermenting sugar molecules to a product. Additionally, enzymatic and/or proteinaceous extracts may be isolated from one or more genetically modified organisms. The extracts are utilized to convert biomass into a product. Further provided are methods of converting biomass into products comprising: placing the genetically modified organism and/or enzymatic extracts thereof in fluid contact with polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, and/or xylan-, glucan-, galactan-, or mannan-decorating groups.
Idaho National Laboratory 01/09/2018
Issued
Patent 8,716,011
Patent
8,716,011
Transcriptional control in Alicyclobacillus acidocaldarius and associated genes, proteins, and methods
Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods of modulating transcription or transcription or transcriptional control using isolated and/or purified polypeptides and nucleic acid sequences from Alicyclobacillus acidocaldarius.
Idaho National Laboratory 05/06/2014
Issued
Patent 8,969,033
Patent
8,969,033
Alteration and modulation of protein activity by varying post-translational modification
Embodiments of the invention include methods of altering the enzymatic activity or solubility of an extremophilic enzyme or post-translationally modifying a protein of interest via using isolated or partially purified glycosyltransferases and/or post-translational modification proteins, extracts of cells comprising glycosyltransferases and/or post-translational modification proteins, and/or in cells comprising one or more glycosyltransferases and/or post-translational modification proteins.
Idaho National Laboratory 03/03/2015
Issued
Patent 9,234,228
Patent
9,234,228
Thermophilic and thermoacidophilic glycosylation genes and enzymes from Alicyclobacillus acidocaldarius and related organisms, methods
Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods for glycosylating and/or post-translationally modifying proteins using isolated and/or purified polypeptides and nucleic acid sequences from Alicyclobacillus acidocaldarius.
Idaho National Laboratory 01/12/2016
Issued
Patent 8,492,114
Patent
8,492,114
Methods of combined bioprocessing and related microorganisms, thermophilic and/or acidophilic enzymes, and nucleic acids encoding said enzymes
A genetically modified organism comprising: at least one nucleic acid sequence and/or at least one recombinant nucleic acid isolated from Alicyclobacillus acidocaldarius and encoding a polypeptide involved in at least partially degrading, cleaving, transporting, metabolizing, or removing polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups; and at least one nucleic acid sequence and/or at least one recombinant nucleic acid encoding a polypeptide involved in fermenting sugar molecules to a product. Additionally, enzymatic and/or proteinaceous extracts may be isolated from one or more genetically modified organisms. The extracts are utilized to convert biomass into a product. Further provided are methods of converting biomass into products comprising: placing the genetically modified organism and/or enzymatic extracts thereof in fluid contact with polysaccharides, cellulose, lignocellulose, hemicellulose, lignin, starch, sugars, sugar oligomers, carbohydrates, complex carbohydrates, chitin, heteroxylans, glycosides, and/or xylan-, glucan-, galactan-, or mannan-decorating groups.
Idaho National Laboratory 07/23/2013
Issued
Patent 8,728,803
Patent
8,728,803
Thermophilic and thermoacidophilic metabolism genes and enzymes from Alicyclobacillus acidocaldarius and related organisms, methods
Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods for modulating or altering metabolism in a cell using isolated and/or purified polypeptides and nucleic acid sequences from Alicyclobacillus acidocaldarius.
Idaho National Laboratory 05/20/2014
Issued
Patent 7,858,353
Patent
7,858,353
Thermophilic and thermoacidophilic biopolymer-degrading genes and enzymes from alicyclobacillus acidocaldarius and related organisms, methods
Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods of at least partially degrading, cleaving, or removing polysaccharides, lignocellulose, cellulose, hemicellulose, lignin, starch, chitin, polyhydroxybutyrate, heteroxylans, glycosides, xylan-, glucan-, galactan, or mannan-decorating groups using isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius.
Idaho National Laboratory 12/28/2010
Issued
Patent 8,569,030
Patent
8,569,030
Type II restriction-modification system methylation subunit of Alicyclobacillus acidocaldarius
Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods for modulating or altering recombination inside or outside of a cell using isolated and/or purified polypeptides and/or nucleic acid sequences from Alicyclobacillus acidocaldarius.
Idaho National Laboratory 10/29/2013
Issued
Technology Status
Technology IDDevelopment StageAvailabilityPublishedLast Updated
BA-062, BA-102DevelopmentAvailable08/01/201808/01/2018

Contact INL About This Technology

To: Ryan Bills<Ryan.Bills@inl.gov>