Multimeric protein for fermentation of ethanol from pla nt biomass; enhancing quality of animal feeds
Inventors
BAEZ-VASQUEZ, MARCO A. • Burlingame, Richard • Magnuson, Jon K. • Bradford, Marion • Sinitsyn, Arkady Panteleimonovich
Assignees
Battelle Memorial Institute Inc • Dyadic International Inc • Iowa Corn Promotion Board
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Publication Number
US-8304212-B2
Publication Date
2012-11-06
Expiration Date
Abstract
Methods to convert lignocellulosic biomass to fermentable sugars with enzymes that degrade the lignocellulosic material are provided, as well as novel combinations of enzymes, including those that provide a synergistic release of sugars from plant biomass.
Core Innovation
The invention relates to a multi-enzyme product comprising isolated enzymes from Trichoderma that exhibits defined minimum enzymatic activities, including glucoamylase activity, β-xylosidase activity, β-glucosidase activity, and α-arabinofuranosidase activity. The multi-enzyme product is characterized as increasing the yield of fermentable sugars from Distiller’s Dried Grains (DDG) compared with compositions not including the claimed amount of glucoamylase.
The invention addresses saccharification of lignocellulosic biomass and describes multi-enzyme products derived from fungi such as Chrysosporium, Trichoderma, Aspergillus, and Penicillium/Talaromyces. Synergistic multi-enzyme products are used to convert lignocellulosic material and plant biomass into fermentable sugars, including in the context of DDG/DDGS derived from corn dry milling byproducts.
The disclosure provides multi-enzyme products that are optionally provided as crude fermentation products or after purification, and may include accessory enzymes. It further describes methods of contacting lignocellulose or DDG with the multi-enzyme products, fermentation of the resulting sugars to produce organic substances including ethanol, and production of the multi-enzyme products by cultivating fungi or using genetically modified organisms overexpressing relevant enzymes. It also describes using recovered saccharified solids as animal feed additives to improve nutritional quality.
The invention includes experimental support showing DDG hydrolysis using fungal enzyme preparations, analysis of sugar composition including HPLC, and observed synergy of enzyme mixtures, including benefit of heat pretreatment in improving saccharification. It also describes screening for efficient enzyme sources and factors affecting saccharification, including phenolics, enzyme ratios, and relief of β-glucosidase inhibition.
Claims Coverage
The independent claim covers a Trichoderma-derived multi-enzyme product with specified minimum enzyme activity thresholds and a functional requirement to increase fermentable sugar yield from DDG relative to compositions that do not include the claimed glucoamylase amount. Dependent claims refine the product form, processing, and the substrate/source scope by narrowing specific fungal species and the DDG feedstock origin.
Trichoderma-derived multi-enzyme product with specified minimum enzyme activities
A multi-enzyme product comprising isolated enzymes from a Trichoderma and exhibiting: a glucoamylase activity of at least about 0.001 U per milligram of dry matter, a β-xylosidase activity of at least about 280 nmol/min/mg of dry matter, a β-glucosidase activity of at least about 454 nmol/min/mg of dry matter, and an α-arabinofuranosidase activity of at least about 100 nmol/min/mg of dry matter.
Increased fermentable sugar yield from DDG relative to compositions lacking the claimed glucoamylase amount
The multi-enzyme product increases the yield of fermentable sugars from Distiller's Dried Grains (DDG) over compositions not including the claimed amount of glucoamylase.
Multi-enzyme product characterized by Trichoderma reesei
The multi-enzyme product is characterized by having Trichoderma as Trichoderma reesei.
Multi-enzyme product having a purification step
The multi-enzyme product has undergone a purification step.
Multi-enzyme product characterized as a crude fermentation product
The multi-enzyme product is a crude fermentation product.
Multi-enzyme product capable of converting lignocellulosic material to sugars
The multi-enzyme product is capable of converting lignocellulosic material to sugars.
DDG derived from corn
The DDG is derived from corn.
Overall, the claim set is directed to a Trichoderma isolated-enzyme multi-enzyme product meeting minimum activity thresholds for glucoamylase, β-xylosidase, β-glucosidase, and α-arabinofuranosidase, with a performance requirement to increase fermentable sugar yield from DDG versus compositions lacking the claimed glucoamylase level. Dependent claims further define the product source as Trichoderma reesei, product processing as crude fermentation product or purification, and substrate/feedstock limitations such as corn-derived DDG, and define capability for converting lignocellulosic material to sugars.
Stated Advantages
Increases the yield of fermentable sugars from Distiller’s Dried Grains (DDG) over compositions not including the claimed amount of glucoamylase.
Documented Applications
Saccharification of lignocellulosic biomass, including DDG/DDGS, to produce fermentable sugars for fermentation and production of organic substances including ethanol.
Use of recovered saccharified solids as animal feed additives to improve nutritional quality.
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