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WO1997010352A1 - Traitement de materiaux cellulosiques par des bacteries productrices de cellulase - Google Patents

Traitement de materiaux cellulosiques par des bacteries productrices de cellulase Download PDF

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Publication number
WO1997010352A1
WO1997010352A1 PCT/US1995/011597 US9511597W WO9710352A1 WO 1997010352 A1 WO1997010352 A1 WO 1997010352A1 US 9511597 W US9511597 W US 9511597W WO 9710352 A1 WO9710352 A1 WO 9710352A1
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atcc
microorganism
cellulase
mutant strain
identifying characteristics
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PCT/US1995/011597
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English (en)
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H. Craig Dees
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Lockheed Martin Energy Systems, Inc.
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Publication date
Application filed by Lockheed Martin Energy Systems, Inc. filed Critical Lockheed Martin Energy Systems, Inc.
Priority to AU36308/95A priority Critical patent/AU3630895A/en
Priority to PCT/US1995/011597 priority patent/WO1997010352A1/fr
Publication of WO1997010352A1 publication Critical patent/WO1997010352A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/01—Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • D—TEXTILES; PAPER
    • D21—PAPER-MAKING; PRODUCTION OF CELLULOSE
    • D21C—PRODUCTION OF CELLULOSE BY REMOVING NON-CELLULOSE SUBSTANCES FROM CELLULOSE-CONTAINING MATERIALS; REGENERATION OF PULPING LIQUORS; APPARATUS THEREFOR
    • D21C5/00—Other processes for obtaining cellulose, e.g. cooking cotton linters ; Processes characterised by the choice of cellulose-containing starting materials
    • D21C5/005—Treatment of cellulose-containing material with microorganisms or enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/38—Pseudomonas

Definitions

  • the present invention relates generally to the conversion of cellulosic materials into fuels, sugars, and specialty chemicals, and more particularly to bacteria which produce cellulase and other enzymes under various environmental conditions.
  • Waste disposable cellulosic materials i.e., newsprint, diapers, packaging, containers, etc.
  • Cellulose is generally defined as a polymer of ⁇ -D-glucose units.
  • Organisms with enhanced cellulase producing capabilities that can tolerate alkaline conditions are advantageous for use in bioreactors for converting cellulose waste or fossil fuels.
  • Most efforts on cellulose degrading microorganisms has centered on the cellulose degrading fungi (i.e., Trichoderma reesei).
  • filamentous fungi are more difficult to grow in a fermentor than a bacterial cellulose degrader.
  • Fungi are more difficult than bacteria to genetically engineer (utilizing recombinant DNA methods) to enhance cellulase production or to modify metabolic capabilities.
  • Most efforts on bacterial cellulases have been concentrated on the anaerobic cellulose degrading bacteria.
  • RIF ⁇ TS OF THF INVFNTI N Accordingly, it is an object of the present invention to provide a new and improved bacterium with increased enzyme producing ability. It is another object of the present invention to provide a new and improved bacterium with increased cellulase producing ability. It is another object of the present invention to provide a new and improved bacterial cloning host with increased cellulase producing ability. It is another object of the present invention to provide a new and improved bacterium with increased enzyme producing ability over a wide pH range. It is another object of the present invention to provide a new and improved bacterium with increased cellulase producing ability over a wide pH range.
  • a microorganism consisting of ATCC 55703, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism.
  • a microorganism consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism.
  • the foregoing and other objects are achieved by: a cellulase produced by a microorganism consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism.
  • a method for producing a cellulase which comprises culturing a cellulase-producing microorganism in a medium, the microorganism consisting of ATCC 55703, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microor ' T iism.
  • a method for producing a cellulase which comprises culturing a cellulase-producing microorganism in a medium, the microorganism consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism.
  • a composition comprising: a cellulase produced by a microorganism consisting of ATCC 55703, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism; and a cellulosic material.
  • a composition comprising: a cellulase produced by a microorganism consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism; and a cellulosic material.
  • a method of treating a cellulosic material comprising: contacting the cellulosic material with an amount of a cellulase produced by a microorganism consisting of ATCC 55703, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism.
  • a method of treating a cellulosic material comprising: contacting the cellulosic material with an amount of a cellulase produced by a microorganism consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism.
  • a detergent comprising: a cellulase produced by a microorganism consisting of ATCC 55703, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism; a surfactant; and a builder.
  • a detergent comprising: a cellulase produced by a microorganism consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of the microorganism; a surfactant; and a builder.
  • the foregoing and other objects are achieved by: a bacteria] cloning host consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of said bacterial cloning host.
  • a bacteria] cloning host consisting of ATCC 55702, or a mutant strain thereof possessing substantially all of the identifying characteristics of said bacterial cloning host.
  • an enzyme produced by a microorganism consisting of ATCC 55703, or a mutant strain thereof possessing substantially all of the identifying characteristics of said microorganism.
  • Figure 1 illustrates a temperature growth curve for the bacteria corresponding to ATCC 55703 and ATCC 55702, in accordance with one aspect of the present invention.
  • Figure 2 illustrates a pH stability curve for the bacteria corresponding to ATCC 55703 and ATCC 55702, in accordance with one aspect of the present invention.
  • Figure 3 illustrates a salt tolerance growth curve for the bacteria corresponding to ATCC 55703 and ATCC 55702, in accordance with one aspect of the present invention.
  • Figure 4 illustrates a comparison of cellulase (CMCase) activity in culture supernatants of the bacterium corresponding to ATCC 55702 compared to the bacterium corresponding to ATCC 55703, in accordance with one aspect of the present invention.
  • Figure 5 illustrates an ultraviolet kill curve for a rec- equivalent cellulose degrading bacterium corresponding to ATCC 55702, as compared to an E. coli rec- and ATCC 55703, in accordance with one aspect of the present invention.
  • microorganisms that are capable of producing cellulases have increased with new demands for industrial processes to degrade or modify cellulosic biomass to commercially important materials.
  • the enzymes produced by those microorganisms are not only important in conversion of cellulosic waste to commercially useful feed stocks but also to degrade cellulose and hemicellulose to materials that can be used in the production of alternative fuels.
  • a wide variety of anaerobic and aerobic bacterial cellulase producers have been described including the bacterium NCIB 10462. This organism was originally named Pseudomonas ⁇ uorescens var. cellulosa.
  • the bacterium has also been called Pseudomonas ⁇ uorescens subsp. cellubsa in recent publications. This bacterium was isolated from soil in the early 1950' s and was described by Ueda. The bacterium was placed in the National Council for Industrial Bacteriology (hereinafter referred to as NCIB) repository in Scotland, and was given the designation "NCIB 10462.” Interest in the study of NCIB 10462 has recently increased along with the recent upsurge of interest in biotechnological conversion of cellulose. When a sample of NCIB 10462 was recently examined, it was found to have very few identifying characteristics of Pseudomonas fluorescens var.
  • Pseudomonas cellulosa #28 was deposited in the American Type Culture Collection (hereinafter referred to as ATCC) and was given Accession No. 55703 (hereinafter referred to as ATCC 55703).
  • ATCC 55703 was then subjected to intense genetic modification using mutagenesis by N-methyl-N'-nitro-N-Nitrosoguanidine (hereinafter referred to as MNNG) treatment.
  • MNNG N-methyl-N'-nitro-N-Nitrosoguanidine
  • the protocol for this mutagenesis treatment is fully explained in F.xppriment ⁇ in nlffnilar Ot-.np.tir-;, Miller, pp 125-129 (1972), the entire disclosure of which is incorporated herein by reference.
  • MNNG was utilized because it induces a high frequency of mutations at doses which result in little killing.
  • the resulting microorganism was then designated as Pseudomonas species #142 (or Pseudomonas sp. #142).
  • ATCC 55702 This microorganism was then deposited in the ATCC and was given Accession No. 55702 (hereinafter referred to as ATCC 55702). Due to the new importance of these novel bacteria as candidates for commercial biomass conversion, the metabolic and physical properties of ATCC 55703 and ATCC 55702 were examined. ATCC 55703 and ATCC 55702 were examined by utilizing a wide variety of standard bacteriological methods which included assimilation studies, fatty acid analysis, and lipid analysis. ATCC 55703 and ATCC 55702 were found to grow aerobically on a wide variety of solid and soluble cellulosic materials. ATCC 55703 and ATCC 55702 grew in complex medium under reducing conditions and weakly on complex media under full anaerobic conditions.
  • ATCC 55703 and ATCC 55702 were found to grow best aerobically but also grew well in complex media under reducing conditions. ATCC 55703 and ATCC 55702 grew slowly under full anaerobic conditions on complex media and did not grow anaerobically on cellulose media as the sole carbon source.
  • Total fatty acid analysis (MIDI) of ATCC 55703 and ATCC 55702 failed to group these bacteria with any known genus or species (very low probability match with the genus Vibrio).
  • ATCC 55703 and ATCC 55702 will continue to be designated as belonging to the Pseudomonas genus.
  • ATCC 55703 and ATCC 55702 were maintained on solid media consisting of M9 liquid medium to which 15 grams per liter agarose had been added along with 0.1 % carboxymethyl cellulose (CMC) as the sole carbon source.
  • CMC carboxymethyl cellulose
  • ATCC 55703 and ATCC 55702 were then transferred to Trypticase soy agar prior to MIDI analysis. Fatty acid analysis by the MIDI system was performed on ATCC 55703 and ATCC 55702 growing anaerobically on 5% sheeps blood agar.
  • Cellulosic media consisted of M9 salt solution as previously described. Agar was added to 15 grams per liter for solid media along with soluble cellulosic component (i.e., carboxymethyl cellulose (CMC)) to 0J % (w/v). Cellulosic liquid media consisted of M9 salt solution to which strips of filter paper or newspaper had been added. Alternatively, Avicell (powder) was added to 0.1% (w/v). All bacteriologic assays and growth procedures were performed using standard bacteriology techniques, methods and medias. Non-fermentative analysis was performed using an API non-fermentor identification system.
  • CMC carboxymethyl cellulose
  • ATCC 55703 and ATCC 55702 are small non-fermentative gram negative rod bacteria with few characteristics that are typically associated with P. fluorescens or even with the genus Pseudomonas.
  • Table 1 illustrates the similarities and differences between ATCC 55703 and ATCC 55702 with respect to P. fluorescens regarding standard biochemical assays:
  • Table 2 illustrates various mo ⁇ hological and biochemical characteristics of both ATCC 55703 and ATCC 55702:
  • ATCC 55703 and ATCC 55702 can utilize a wide variety of solid and soluble cellulosic media as a sole carbon source which is consistent with previous observations.
  • ATCC 55703 and ATCC 55702 are oxidase and catalase positive, motile, and aerobic, which are the few characteristics that suggest it might be related to the Pseudomonads.
  • ATCC 55703 and ATCC 55702 only weakly clot a ly ulus lysate test which is interesting since they appear to be 0.5-1 ⁇ m gram negative rods using gram stain. These studies show that ATCC 55703 and ATCC 55702 are not closely related to the genus Pseudomas, but may have a very low probability match to the genus Sphingomonas.
  • ATCC 55703 and ATCC 55702 have an operating temperature range of 5-37°C, a preferred temperature range of 25-32°C, and an optimal temperature of 30°C. No growth of ATCC 55703 or ATCC 55702 has been observed at 45°C.
  • the operating temperature range is 5-70°C (at pH 7.5).
  • the preferred temperature is 55°C (at pH 7.5).
  • ATCC 55703 and ATCC 55702 have an operating pH range of 6.8 to 9.0, a preferred pH range of 7.2 to 8.0, with growth decreasing after the pH is adjusted below pH 7.0.
  • ATCC 55703 and ATCC 55702 reduced the medium pH below pH 6.8, whereas in liquid cellulosic medium the pH does not fall more than 0.3 from the starting pH of 7.2.
  • the operating pH range is 5-11 (at 55°C).
  • the preferred pH range is 7-8 (at 55°C).
  • the salt tolerance curve illustrates that ATCC 55703 and ATCC 55702 have increased tolerance to twice as much sodium than is commonly used in growth medium.
  • ATCC 55703 and ATCC 55702 have an operating salt range of 8.5-32 mM sodium, with a preference of 8.5 mM sodium. Less dilution of high salt brines used in paper processing will be required to use ATCC 55703 and ATCC 55702 to biodegrade cellulosic waste from the paper industry. Taxonomic characterization of ATCC 55703 and ATCC 55702 using total fatty acid analysis failed to group these bacteria with any of the currently recognized genera. Since the non-fermentative analysis suggested that ATCC 55703 and ATCC 55702 may be distantly related members of the genus Sphingomonas, a total lipid analysis was also performed on ATCC 55703 and ATCC 55702.
  • the polar lipid fractions were comprised of ester-linked acyl- glycerophophatides, which would rule out classification of ATCC 55703 or ATCC 55702 in the genus Sphingomonas.
  • Total fatty acid profiling has been used to identify anaerobic and aerobic bacteria.
  • Total fatty acid analysis failed to adequately group ATCC 55703 or ATCC 55702 into any known genus. Only a low probability match with the genus Vibrio was obtained.
  • Total fatty acid analysis found that a large percentage of the fatty acids produced by ATCC 55703 and ATCC 55702 are in the trans configurations which is characteristic of some members of Vtbrio.
  • ATCC 55703 and ATCC 55702 are capable of degrading a wide variety of soluble and solid cellulosic materials. Specifically, ATCC 55703 and ATCC 55702 have improved characteristics and qualities for the degradation of cellulosic waste, degradation of cellulosic fossil fuels (i.e., coal) for alternative fuel production, detergents, drain pipe cleaners, septic tank cleaners/additives, textile processing, and paper processing. However, ATCC 55702 has demonstrated a far greater ability to produce large amounts of cellulase than ATCC 55703.
  • Figure 4 shows that the production of extracellular cellulase from ATCC 55702 is approximately 300 times higher than ATCC 55703. After 24 hours, the zone of clearing around the well of ATCC 55702 is very large (upper right hand corner) (arrow) compared to ATCC 55703 or eleven other cellulase degrading bacteria (no zones visible).
  • ATCC 55702 It has also been found that for ATCC 55702 to grow rapidly on solid cellulose and produce the highest amounts of extracellular endocellulase, it must also demonstrate high levels of ⁇ -glucosidase (cellobiase) activity. This enzyme converts cellobiose to glucose. ATCC 55702 rapidly produces a colored product from X-GLU (a synthetic marker/test for ⁇ -glucosidase production). Therefore, ATCC 55702 is enhanced not only in the production of extracellular cellulases (endoglucanases), but is also enhanced for ⁇ -glucosidase activity.
  • X-GLU a synthetic marker/test for ⁇ -glucosidase production
  • ATCC 55702 is especially suited to biomass conversion of cellulosic waste for alternative fuel production.
  • ATCC 55702 grows best and produces cellulases at temperatures lower than 37°C, a temperature most often used to grow most bacteria and fungi. Lower temperature requirements means that the yield from biomass conversion by ATCC 55702 is more suitable for energy production.
  • the energy required to produce the fuel by growing ATCC 55702 at 20-24°C is much less than heating the fermentor or bioreactor to 37°C.
  • ATCC 55702 is greatly superior for alternative fuel production as compared to the thermophilic cellulose degrading bacteria whose growth requires temperatures ranging from 45-65°C.
  • a simple powder containing the cellulase and bacteria can be used as an additive to cellulosic waste (i.e., diapers, tampons, newsprint, etc.).
  • the product would be embodied as an inexpensive dried or lyophilized powder of the total fermentate which contains bacteria and cellulase. No post fermentation ("down stream") processing would be required. The product would only require fermentation, lyophilization or drying, and packaging.
  • the powder could consist alternatively of: (1) dried fermentate (cellulase) plus cells (bacteria); (2) dried cells (bacteria); or (3) dried fermentate (cellulase).
  • the product could be packaged in a liquid format.
  • the liquid product could consist alternatively of: (1) fermentate (cellulase); (2) lysed cells (bacteria); or (3) fermentate (cellulase) plus lysed cells (bacteria).
  • any trait introduced into a bacterium by either classical methods (mutagenesis) or recombinant DNA technologies (genetic engineering) can be destroyed by rearrangement of the genetic material by the DNA enzymes.
  • mutagenesis classical methods
  • recombinase deficient cloning hosts have been created that lack these recombinase genes ⁇ rec- strains).
  • cloning of cellulase genes into E. coli is not commercially practical since the organism cannot secrete the cellulase.
  • ATCC 55702 would therefore be an improved host for genetic manipulations using recombinant DNA techniques, and is less likely to destroy genetic manipulations using standard mutagenesis techniques.
  • ATCC 55702 is equivalent to Escherichia coli recombinant deficient ⁇ rec-).
  • FIG. 5 illustrates a UV kill curve performed on ATCC 55703, which is UV resistant.
  • a rec- E. coli cloning host is rapidly killed by slow exposure to UV radiation ( Figure 5).
  • the kill curve of ATCC 55702 (a UV sensitive ⁇ rec- equivalent) cellulose degrading bacterium) is shown in Figure 5. Therefore, the ATCC 55702 cloning host will be less able to recombine/repair DNA as efficiently as the wild type bacterium. This feature limits the cloning host from "moving" the genetic elements and destroying them via DNA repair or recombination mechanisms.
  • ATCC 55703 was resistant to ampicillin and was partially resistant to tetracycline. Selection via replicate plating during the process of developing the ATCC 55702 rec- strain produced a strain that was also highly sensitive to tetracycline and ampicillin. Therefore, in accordance with one aspect of the present invention, the ATCC 55702 cloning host is highly sensitive to these two antibiotics, which is illustrated in Table 5, below:
  • ATCC 55703 was subjected to an intensive mutagenesis regimen using MNNG. Resulting strains were selected for: (1) improved soluble cellulase production; (2) rapid degradation of solid cellulose and waste products (filter paper, newsprint and diapers); (3) UV sensitivity; (4) enhanced ⁇ -glucosidase activity; (5) tetracycline and ampicillin sensitivity; and (6) high levels of total carbohydrate production from solid cellulose. As a result of replicate plating procedures, ATCC 55702 was identified.
  • the present invention has many applications with respect to detergents, drain pipe cleaners, septic tank cleaners additives, conversion of cellulosic materials to animal feedstocks, conversion of starches to sugars, increased lignin and hemicellulose removal from cellulosic materials, textiles, food processing, enzymes, detergents, paper products, bioremediation, specialty chemicals, consumer goods, and renewable fuels.
  • Th ⁇ iis sheet For receiving Office use only For International Bureau use only ⁇ Th ⁇ iis sheet was received with the international application I I This sheet was received by the International Bureau on:

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Abstract

Des bactéries produisant de grandes quantités de cellulase, ainsi que d'autres enzymes, ont été identifiées. La bactérie d'origine (ATCC 55703) a été génétiquement modifiée à l'aide d'un traitement par nitrosoguanidine (MNNG, de façon à donner la bactérie améliorée productrice de cellulase (ATCC 55702), qui a été identifiée par mise en culture pour réplication. L'ATCC 55702 présente de meilleures caractéristiques et qualités pour la dégradation des déchets cellulosiques en vue de la production de combustibles, du traitement des aliments, du traitement des textiles et d'autres applications industrielles. L'ATCC 55702 est un hôte bactérien amélioré pour les manipulations génétiques faisant appel à des techniques de recombinaison de l'ADN et il est moins susceptible de détruire les manipulations génétiques faisant appel à des techniques de mutagenèse standard.
PCT/US1995/011597 1995-09-14 1995-09-14 Traitement de materiaux cellulosiques par des bacteries productrices de cellulase WO1997010352A1 (fr)

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PCT/US1995/011597 WO1997010352A1 (fr) 1995-09-14 1995-09-14 Traitement de materiaux cellulosiques par des bacteries productrices de cellulase

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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7408056B2 (en) 1999-06-22 2008-08-05 Xyleco, Inc. Cellulosic and lignocellulosic materials and compositions and composites made therefrom
WO2009113882A1 (fr) * 2008-03-11 2009-09-17 Institute Of Geological And Nuclear Sciences Limited Micro-organismes
EP2218773A1 (fr) * 2009-02-17 2010-08-18 Deinove Compositions et procédés pour dégrader la biomasse lignocellulosique
EP2251414A1 (fr) * 2009-05-14 2010-11-17 Deinove Bactérie métabolique haute performance
WO2013092645A1 (fr) * 2011-12-19 2013-06-27 Deinove Ingrédients pour compositions d'aliments pour animaux
US9005954B2 (en) 2009-01-19 2015-04-14 Deinove Methods for isolating bacteria
US9034619B2 (en) 2009-05-14 2015-05-19 Deinove Recombinant bacteria and the uses thereof for producing ethanol
US9181564B2 (en) 2007-11-14 2015-11-10 Deinove Use of bacteria for the production of bioenergy
US9279162B2 (en) 2010-03-02 2016-03-08 Deinove Bacteria and the uses thereof
US10059035B2 (en) 2005-03-24 2018-08-28 Xyleco, Inc. Fibrous materials and composites

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US3616222A (en) * 1969-05-08 1971-10-26 Exxon Research Engineering Co Process for saccharification of cellulosic and woody tissues by fungi or enzymes from fungi
US4108724A (en) * 1975-04-11 1978-08-22 Takeda Chemical Industries, Ltd. Method for preparing antibiotic P-2563 using Pseudomonas fluorescens
US4479936A (en) * 1982-09-27 1984-10-30 Microlife Technics, Inc. Method for protecting the growth of plants employing mutant siderophore producing strains of Pseudomonas Putida
US5340731A (en) * 1988-07-08 1994-08-23 University Of British Columbia Method of preparing a B-1,4 glycan matrix containing a bound fusion protein
US5348742A (en) * 1991-05-24 1994-09-20 Ciba-Geigy Corporation Anti-pathogenic bacterial strains of Pseudomonas fluorescens

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3616222A (en) * 1969-05-08 1971-10-26 Exxon Research Engineering Co Process for saccharification of cellulosic and woody tissues by fungi or enzymes from fungi
US4108724A (en) * 1975-04-11 1978-08-22 Takeda Chemical Industries, Ltd. Method for preparing antibiotic P-2563 using Pseudomonas fluorescens
US4479936A (en) * 1982-09-27 1984-10-30 Microlife Technics, Inc. Method for protecting the growth of plants employing mutant siderophore producing strains of Pseudomonas Putida
US5340731A (en) * 1988-07-08 1994-08-23 University Of British Columbia Method of preparing a B-1,4 glycan matrix containing a bound fusion protein
US5348742A (en) * 1991-05-24 1994-09-20 Ciba-Geigy Corporation Anti-pathogenic bacterial strains of Pseudomonas fluorescens

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7537826B2 (en) 1999-06-22 2009-05-26 Xyleco, Inc. Cellulosic and lignocellulosic materials and compositions and composites made therefrom
US7408056B2 (en) 1999-06-22 2008-08-05 Xyleco, Inc. Cellulosic and lignocellulosic materials and compositions and composites made therefrom
US10059035B2 (en) 2005-03-24 2018-08-28 Xyleco, Inc. Fibrous materials and composites
US9181564B2 (en) 2007-11-14 2015-11-10 Deinove Use of bacteria for the production of bioenergy
WO2009113882A1 (fr) * 2008-03-11 2009-09-17 Institute Of Geological And Nuclear Sciences Limited Micro-organismes
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EP2218773A1 (fr) * 2009-02-17 2010-08-18 Deinove Compositions et procédés pour dégrader la biomasse lignocellulosique
JP2012517817A (ja) * 2009-02-17 2012-08-09 ディーノヴ リグノセルロース系バイオマスを分解するための組成物及び方法
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EA024724B1 (ru) * 2009-02-17 2016-10-31 Деинов Способ выделения уф-устойчивых бактерий и их применение
WO2010094665A2 (fr) 2009-02-17 2010-08-26 Deinove Compositions et procédés de dégradation de biomasse ligneuse
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JP2012526538A (ja) * 2009-05-14 2012-11-01 ディーノヴ 高性能代謝細菌
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CN102482637A (zh) * 2009-05-14 2012-05-30 德诺芙公司 高性能代谢细菌
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US9725741B2 (en) 2009-05-14 2017-08-08 Deinove Recombinant bacteria and the uses thereof for producing ethanol
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