News Column

"Processing Biomass" in Patent Application Approval Process

January 30, 2014

By a News Reporter-Staff News Editor at Agriculture Week -- A patent application by the inventors MEDOFF, Marshall (Brookline, MA); MASTERMAN, Thomas Craig (Brookline, MA); YOSHIDA, Aiichiro (Canton, MA); MOON YEE FUNG, Jennifer (Salem, MA); LYNCH, James J. (Woburn, MA), filed on September 3, 2013, was made available online on January 16, 2014, according to news reporting originating from Washington, D.C., by VerticalNews correspondents.

This patent application is assigned to Xyleco, Inc.

The following quote was obtained by the news editors from the background information supplied by the inventors: "As demand for petroleum increases, so too does interest in renewable feedstocks for manufacturing biofuels and biochemicals. The use of lignocellulosic biomass as a feedstock for such manufacturing processes has been studied since the 1970s. Lignocellulosic biomass is attractive because it is abundant, renewable, domestically produced, and does not compete with food industry uses.

"Many potential lignocellulosic feedstocks are available today, including agricultural residues, woody biomass, municipal waste, oilseeds/cakes and sea weeds, to name a few. At present these materials are either used as animal feed, biocompost materials, are burned in a cogeneration facility or are landfilled.

"Lignocellulosic biomass is recalcitrant to degradation as the plant cell walls have a structure that is rigid and compact. The structure comprises crystalline cellulose fibrils embedded in a hemicellulose matrix, surrounded by lignin. This compact matrix is difficult to access by enzymes and other chemical, biochemical and biological processes. Cellulosic biomass materials (e.g., biomass material from which substantially all the lignin has been removed) can be more accessible to enzymes and other conversion processes, but even so, naturally-occurring cellulosic materials often have low yields (relative to theoretical yields) when contacted with hydrolyzing enzymes. Lignocellulosic biomass is even more recalcitrant to enzyme attack. Furthermore, each type of lignocellulosic biomass has its own specific composition of cellulose, hemicellulose and lignin.

"While a number of methods have been tried to extract structural carbohydrates from lignocellulosic biomass, they are either too expensive, produce too low a yield, leave undesirable chemicals in the resulting product, or simply degrade the sugars.

"Saccharides from renewable biomass sources could become the basis of chemical and fuels industries by replacing, supplementing or substituting petroleum and other fossil feedstocks. However, techniques need to be developed that will make these monosaccharides available in large quantities and at acceptable purities and prices."

In addition to the background information obtained for this patent application, VerticalNews journalists also obtained the inventors' summary information for this patent application: "Provided herein are methods of inducing the production of one or more enzymes by a microorganism, through the use of an inductant.

"In one aspect, a method is provided that includes combining a cellulosic or lignocellulosic biomass, which has been treated to reduce its recalcitrance, with a microorganism, to induce the production of one or more enzyme(s) by the microorganism by maintaining the microorganism-biomass combination under conditions that allow for the production of the enzyme(s) by the microorganism. In some implementations, the enzyme(s) are then used to saccharify cellulosic or lignocellulosic biomass.

"Also provided herein is a method for inducing the production of an enzyme by a microorganism, where the method includes: providing a first cellulosic or lignocellulosic biomass; treating the first biomass with a treatment method to reduce its recalcitrance, thereby producing a first treated biomass; providing a microorganism; providing a liquid medium; combining the first treated biomass, the microorganism, and the liquid medium, thereby producing a microorganism-biomass combination; and maintaining the microorganism-biomass combination under conditions allowing for the production of an enzyme by the microorganism, thereby producing an inductant-enzyme combination; thereby inducing the production of the enzyme by the microorganism.

"Also provided herein is a composition that includes a liquid medium, a cellulosic or lignocellulosic biomass treated to reduce its recalcitrance, a microorganism, and one or more enzymes made by the microorganism.

"In any of the methods or compositions provided herein, the treatment for reducing the recalcitrance of the biomass material(s) can be any of: bombardment with electrons, sonication, oxidation, pyrolysis, steam explosion, chemical treatment, mechanical treatment, and freeze grinding. Preferably, the treatment method is bombardment with electrons.

"The methods and compositions can also include mechanically treating the first or the second cellulosic or lignocellulosic biomass to reduce its bulk density and/or increase its surface area. The biomass material(s) can be comminuted before being combined with the microorganism and liquid medium. The comminution can be dry milling or wet milling. The biomass material can have a particle size of about 30 to 1400 .mu.m.

"In any of the methods and compositions described herein, any of the cellulosic or lignocellulosic biomasses can be: paper, paper products, paper waste, paper pulp, pigmented papers, loaded papers, coated papers, filled papers, magazines, printed matter, printer paper, polycoated paper, card stock, cardboard, paperboard, cotton, wood, particle board, forestry wastes, sawdust, aspen wood, wood chips, grasses, switchgrass, miscanthus, cord grass, reed canary grass, grain residues, rice hulls, oat hulls, wheat chaff, barley hulls, agricultural waste, silage, canola straw, wheat straw, barley straw, oat straw, rice straw, jute, hemp, flax, bamboo, sisal, abaca, corn cobs, corn stover, soybean stover, corn fiber, alfalfa, hay, coconut hair, sugar processing residues, bagasse, beet pulp, agave bagasse, algae, seaweed, manure, sewage, offal, agricultural or industrial waste, arracacha, buckwheat, banana, barley, cassava, kudzu, oca, sago, sorghum, potato, sweet potato, taro, yams, beans, favas, lentils, peas, or mixtures of any of these. Alternatively, the cellulosic or lignocellulosic biomass can include material that was remaining after a prior cellulosic or lignocellulosic biomass was previously converted to a product by an enzyme of a microorganism.

"In these methods and compositions, the microorganism can be any of a fungus, a bacterium, or a yeast. The microorganism can actually be a population of different microorganisms. The microorganism can be a strain that produces high levels of cellulase, and/or it can be genetically engineered. The microorganism can be Trichoderma reesei, or it can be Clostridium thermocellum, for example. The microorganism can be a T. reesei strain such as RUT-NG14, PC3-7, QM9414 or RUT-C30.

"In any of these methods and compositions, the cellulosic or lignocellulosic biomass can be combined with the microorganism at a time when the microorganism is in lag phase.

"The methods and compositions can also include removing all or a portion of the liquid from the microorganism-inductant-enzyme combination, to produce an enzyme extract. The methods and compositions can also include concentrating one or more of the enzymes, and/or isolating one or more of the enzymes.

"The methods and compositions can also include allowing saccharification of the second cellulosic or lignocellulosic biomass to occur, so that one or more sugars are produced. The one or more sugars can be isolated and/or concentrated.

"It should be understood that this invention is not limited to the embodiments disclosed in this Summary, and it is intended to cover modifications that are within the spirit and scope of the invention, as defined by the claims.


"The foregoing will be apparent from the following more particular description of example embodiments of the invention, as illustrated in the accompanying drawings in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating embodiments of the present invention.

"FIG. 1 is a diagram illustrating the enzymatic hydrolysis of cellulose to glucose. Cellulosic substrate (A) is converted by endocellulase (i) to cellulose (B), which is converted by exocellulase (ii) to cellobiose (C), which is converted to glucose (D) by cellobiase (beta-glucosidase) (iii).

"FIG. 2 is a flow diagram illustrating conversion of a biomass feedstock to one or more products. Feedstock is physically pretreated (e.g., to reduce its size) (200), optionally treated to reduce its recalcitrance (210), saccharified to form a sugar solution (220), the solution is transported (230) to a manufacturing plant (e.g., by pipeline, railcar) (or if saccharification is performed en route, the feedstock, enzyme and water is transported), the saccharified feedstock is bio-processed to produce a desired product (e.g., alcohol) (240), and the product can be processed further, e.g., by distillation, to produce a final product (250). Treatment for recalcitrance can be modified by measuring lignin content (201) and setting or adjusting process parameters (205). Saccharifying the feedstock (220) can be modified by mixing the feedstock with medium and the enzyme (221).

"FIG. 3 is a flow diagram illustrating the treatment of a first biomass (300), addition of a cellulase producing organism (310), addition of a second biomass (320), and processing the resulting sugars to make products (e.g., alcohol(s), pure sugars) (330). The first treated biomass can optionally be split, and a portion added as the second biomass (A).

"FIG. 4 is a flow diagram illustrating the production of enzymes. A cellulase-producing organism is added to growth medium (400), a treated first biomass (405) is added (A) to make a mixture (410), a second biomass is added (420), and the resulting sugars are processed to make products (e.g., alcohol(s), pure sugars) (430). Portions of the first biomass (405) can also be added (B) to the second biomass (420).

"FIG. 5 shows results of protein analysis using SDS PAGE."

URL and more information on this patent application, see: MEDOFF, Marshall; MASTERMAN, Thomas Craig; YOSHIDA, Aiichiro; MOON YEE FUNG, Jennifer; LYNCH, James J. Processing Biomass. Filed September 3, 2013 and posted January 16, 2014. Patent URL:

Keywords for this news article include: Treatment, Cellulases, Xyleco Inc, Agricultural, Glucosidases, Glycoside Hydrolases, Enzymes and Coenzymes.

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Source: Agriculture Week

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