WO1994004664A1 - New xylanases having high activity and stability at alkaline conditions and high temperatures - Google Patents
New xylanases having high activity and stability at alkaline conditions and high temperatures Download PDFInfo
- Publication number
- WO1994004664A1 WO1994004664A1 PCT/DK1993/000277 DK9300277W WO9404664A1 WO 1994004664 A1 WO1994004664 A1 WO 1994004664A1 DK 9300277 W DK9300277 W DK 9300277W WO 9404664 A1 WO9404664 A1 WO 9404664A1
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- WIPO (PCT)
- Prior art keywords
- enzyme preparation
- enzyme
- strain
- xylanase
- bacillus
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01032—Xylan endo-1,3-beta-xylosidase (3.2.1.32), i.e. endo-1-3-beta-xylanase
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D8/00—Methods for preparing or baking dough
- A21D8/02—Methods for preparing dough; Treating dough prior to baking
- A21D8/04—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes
- A21D8/042—Methods for preparing dough; Treating dough prior to baking treating dough with microorganisms or enzymes with enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/189—Enzymes
-
- 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)
- C12N9/2477—Hemicellulases not provided in a preceding group
- C12N9/248—Xylanases
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01008—Endo-1,4-beta-xylanase (3.2.1.8)
-
- 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
Definitions
- the present invention relates to novel xylanolytic enzymes obtainable from strains of alkalophilic Bacillus sp . Moreover, the invention relates to a method for producing the enzymes and the use of the enzymes in the pulp and paper industry .
- Xylanases with high activity and stability at alkaline conditions are of great commercial interest, e.g. for applications in the pulp and paper industries, for modification of lignocellulose.
- very few xylanases are reported which are able to function at pH values 9-12, and the available literature indicates that these enzymes are rapidly inactivated at a pH of more than 10, especially at temperatures exceeding 50°C.
- the present invention describes new xylanase enzymes obtained from alkaline Bacillus sp.. which are superior to previously described bacterial xylanases with respect to activity and stability in the alkaline region. Furthermore, the xylanases of the present invention are also able to function at high temperature, e.g. 70°C at pH 7-8.
- the invention provides enzyme preparations having xylanolytic activity, and having more than 50% relative activity in the range pH 6-9 at 50°C and temperature optimum in the range of from 55 to 75°C (at pH 6-10) .
- the invention provides a process for the preparation of the enzyme preparations comprising cultivation of a strain of Bacillus sp., preferably the strain Bacillus sp., DSM 7197, or a mutant or a variant thereof, in a suitable nutrient medium, containing carbon and nitrogen sources and inorganic salts, followed by recovery of the desired enzyme.
- the invention relates to the use of the enzyme preparation in a process for treatment of lignocellulosic pulp.
- the invention provides an agent containing an enzyme preparation, provided in the form of a granulate, preferably a non-dusting granulate, a liquid, in particular a stabilized liquid, a slurry, or a protected enzyme.
- Figs. 1-3 show the temperature profiles of the fraction purified according to Ex. 2, in standard Britton & Robinson buffers at pH 7, pH 9, and pH 10, respectively. All reaction mixtures contained 0.013 EXU/ml and were incubated for 30 minutes ( ⁇ sample; A substrate blank);
- Fig. 4 shows the effect of pH on the activity of the fraction purified according to Ex. 2, in 50 mM Britton & Robinson buffers (0.013 EXU/ml; 30 minutes of incubation; 50oC; ⁇ sample; A buffer; ⁇ enzyme blank); and
- Figs. 5, 6, and 7 show the effect of temperature and pH on the stability of the fraction purified according to Ex. 2, in the absence of substrate.
- the fraction was diluted to a concentration of 0.05 EXU/ml in 50 mM Britton & Robinson buffers of pH 7, pH 9, and pH 10, respectively, and incubated at 40°C. At appropriate intervals, 50 ⁇ l samples were removed from each incubation mixtures and transferred to 950 ⁇ l 50 mM Britton & Robinson buffer pH 10. The residual xylanolytic activity was determined at 50°C using Xylazyme TabletsTM (Megazyme, Australia). The incubation time was 30 minutes in all cases ( ⁇ sample; A blind). DETAILED DISCLOSURE OF THE INVENTION
- the invention provides xylanase preparations having high stability and excellent activity at alkaline conditions.
- the enzyme preparation of the invention can be further described by the following characteristics.
- the enzyme preparation of the invention comprises at least 5 xylanolytic enzymes, having pi in the range of from appr. 3 to appr. 9.5.
- the fraction of the enzyme preparation, purified according to Ex. 2 has more than 50% relative activity in the range pH 6-10, determined after 30 minutes of incubation. No pronounced pH optimum is detectable, but appears to be in the range pH 5.5 to 9.0 (cf. Fig. 4).
- the fraction of the enzyme preparation, purified according to Ex. 2 has a temperature optimum in the range of 60 to 75°C, more specifically around 70°C, determined after 30 minutes of incubation (cf. Fig. 1).
- the fraction of the enzyme preparation, purified according to Ex. 2 has a temperature optimum in the range of 55 to 75°C, more specifically in the range of 60 to
- the fraction of the enzyme preparation, purified according to Ex. 2 has a temperature optimum in the range of 50 to 70°C, more specifically around 60°C, determined after 30 minutes of incubation (cf. Fig. 3).
- the fraction of the enzyme preparation, purified according to Ex. 2 has a relative residual activity after incubation for 6 hours at pH 10 and 40°C of at least 90%, more preferred at least 95%, most preferred at least 99%.
- a similar relative residual activity was observed after incubation for 6 hours at pH 10 and 55°C, at pH 10 and 50°C; at pH 10 and 40°C; at pH 9 and 40°C; at pH 9 and 50°C; and at pH 7 and 50°C, cf.
- the enzyme preparation of the invention has immunochemical properties identical or partially identical (i.e. at least partially identical) to those of a xylanase derived from the strain Bacillus sp., DSM 7197.
- the immunochemical properties can be determined immunologically by cross-reaction identity tests.
- the identity tests can be performed by the well-known Ouchterlony double immunodiffusion procedure or by tandem crossed immunoelectrophoresis according to Axelsen N.H.; Handbook of Immunoprecipitation-in-Gel Techniques; Blackwell Scientific Publications (1983), chapters 5 and 14.
- the terms "antigenic identity” and "partial antigenic identity” are described in the same book, chapters 5, 19 and 20.
- Monospecific antiserum was generated according to the above-mentioned method by immunizing rabbits with the purified xylanase of the invention.
- the immunogen was mixed with Freund's adjuvant and injected subcutaneously into rabbits every second week.
- Antiserum was obtained after a total immunization period of 8 weeks, and immunoglobulin was prepared therefrom as described by Axelsen N.H.. supra.
- the enzyme preparations are obtainable by cultivation of alkalophilic Bacillus sp. in a suitable nutrient medium, containing carbon and nitrogen sources and inorganic salts, followed by recovery of the desired enzyme.
- the enzyme preparations are obtained by cultivation of the alkalophilic species described as Group 3 by Gordon & Hyde [Gordon R.E and Hyde J.L. (1982); Journal of General Microbiology, 128 1109-1116, Table
- the enzyme preparations are obtained by cultivation of a strain of the alkalophilic species represented by the strain Bacillus sp.. DSM 7197. In a further preferred embodiment, the enzyme preparations are obtained by cultivation of the strain Bacillus sp., DSM 7197, or a mutant or a variant thereof.
- the enzyme can also be obtained by recombinant DNA-technology.
- the strain Bacillus sp.. DSM 7197 was deposited on 4 August 1992 according to the Budapest Treaty on the International Recognition of the Deposits of Microorganisms for the Purpose of Patent Procedures, at Deutsche Sammlung von Mikroorganismen und Zellkulturen, Mascheroder Weg lb, 3300 Braunschweig, Germany.
- the xylanolytic activity is measured in endo-xylanase units (EXU), determined at pH 9.0 with remazol-xylan as substrate.
- EXU endo-xylanase units
- a xylanase sample is incubated with remazol-xylan substrate.
- the background of non-degraded dyed substrate is precipitated by ethanol.
- the remaining blue colour in the supernatant is proportional to the xylanase activity, and the xylanase units are then determined relatively to an enzyme standard at standard reaction conditions, i.e. at 50.0 +/-0.1°C, pH 9.0, and 30 minutes' reaction time.
- the absorbance of the filtrate is measured at 590 nM. Blank incubations were run in all cases in order to correct for chemical hydrolysis of AZCL-xylan (cf. also Megazyme Product Information Leaflet).
- the invention relates to a method for enzymatic treatment of lignocellulosic pulp, comprising employment of an enzyme of this invention.
- Enzymatic treatment of lignocellulosic pulp improves the bleachability of the pulp and/or reduces the amount of chemicals necessary for obtaining a satisfactory bleaching.
- the enzyme of the invention may also be applied in a complexing stage of the pulp process, prior to hydrogen peroxide or ozone bleaching.
- the xylanase should preferably be provided in the form of a granulate, preferably a non-dusting granulate, a liquid, in particular a stabilized liquid, a slurry, or a protected enzyme.
- the agent contains the xylanase in amounts of at least 20%, preferably at least 30%, of the total enzyme protein.
- the xylanolytic activity can be measured in xylanase units.
- two kinds of units are used: FXU and EXU.
- AF 293.6/1 FXU
- EXU AF 293.9/1
- FXU is determined at pH 6.0
- EXU is determined at pH 9.0.
- FXU and EXU express enzymatic activity in the same order of magnitude.
- the folders AF 293.6/1 and 293.9/1 are available upon request to Novo Nordisk A/S, Denmark, which folders are hereby included by reference.
- the process of the invention is performed at temperatures between 40 and 100oC, more preferred between 50 and 90°C, most preferred between 60 and 80°C.
- the enzymatic treatment is performed at a pH above 5.0, more preferred above 6.0, most preferred above 7.0.
- the enzymatic treatment is performed within a period of 5 minutes to 24 hours, more preferred within 15 minutes to 6 hours, most preferred within 20 minutes to 3 hours.
- a suitable xylanase dosage will usually correspond to a xylanase activity of 10 to 5000 FXU/kg or EXU/kg dry pulp, more preferred 100 to 5000 FXU/kg or EXU/kg dry pulp.
- the enzymatic treatment takes place at a consistency of 3-35%, more preferred 5-25%, most preferred 8-15%.
- the consistency is the dry matter content of the pulp. A pulp with a consistency above 35% is difficult to mix effectively with the enzyme preparation, and a pulp with a consistency below 3% carries too much water, which is a disadvantage from an economic point of view.
- the xylanases of this invention can be implemented in processes for treatment of lignocellulosic pulp essentially as described in e.g. International Patent Application PCT/DK91/00239, or International Patent Publication WO 91/02839.
- the new xylanase enzymes according to the invention may also be well suited for use as baking agents and as additives to animal fodder as described in EP 0 507 723. They may especially be useful for addition to animal feeds for in vivo breakdown of the pentosan fraction as the pH in the small intestine of e.g. poultry, piglets and pigs typically will be in the area of 5.5 to 7 in which area the new xylanase enzymes have significant activity.
- the strain Bacillus sp., DSM 7197 was cultivated at 40°C on a rotary shaking table (300 r.p.m.) in 500 ml baffled Erlenmeyer flasks containing 100 ml of medium of the following composition (per litre):
- the medium is sterilized by heating at 120 "C for 45 minutes .
- the pH of the medium is adjusted to 10.0 by addition of approx. 10 ml 1 M sodium sesquicarbonate to each flask.
- a fraction of xylanases having acidic pl was partially purified by conventional purification techniques involving sample concentration by ultrafiltration and ammonium sulfate precipitation, and conventional chromatographic separation by ionexchange chromatography on S-Sepharose High Load and Q- Sepharose High Load, size exclusion chromatography on Superdex 200 or G-2000 SW, as well as affinity chromatography for specific removal of proteinases.
- the partially purified enzyme fraction obtained according to Ex. 2, was subjected to kinetic studies, and the activity was found to be linear (zero-order kinetics) for at least 6 hours, when incubated at the conditions stated in Table 1, below.
- the partially purified enzyme fraction obtained according to Ex. 2, was subjected to experiments concerning the effects of temperature and pH on enzyme activity and stability using AZCL-xylan tablets (Xylazyme TabletsTM, provided by Megazyme, Australia).
- the assay was performed as follows:
- the above described fraction was diluted to a concentration of 0.05 EXU/ml in 50 mM Britton & Robinson buffers of pH 7, pH 9, and pH 10, respectively, and incubated at 40°C. At appropriate intervals, 50 ⁇ l samples were removed from each incubation mixture and transferred to 950 ⁇ l 50 mM Britton & Robinson buffer pH 10. The residual xylanolytic activity was determined at 50°C using Xylazyme TabletsTM. The incubation time was 30 minutes in all cases. The results are presented in Figs. 5-7. EXAMPLE 5
- N-terminal amino acid sequences of the xylanases were determined using standard methods for obtaining and sequencing peptides (Findlay & Geisow (Eds.), Protein Sequencing - a Practical approach. 1989, IRL Press).
- This amino acid sequence is identical to the amino acid sequence of residues 18 to 29 in a 45 kDa xylanase from the alkalophilic Bacillus sp. C-125 (Hamamoto et al., Agric. Biol. Chem. 51, 1987, pp. 953-955).
- N-terminal amino acid sequence of another xylanase purified from the fermentation broth of ex. 1 by conventional chromatographic methods and characterized by having a MW of approx. 22 kDa using SDS-PAGE and a pl value of approx. 9 in a 3.5 to 9.5 isoelectric focusing gel was found to be (SEQ ID No.2 of the attached sequence listing):
- Asn-Thr-Tyr-Trp-Gln-Tyr-Xaa-Thr-Asp-Gly-Gly-Thr-Val-Asn-Ala-Xaa-Asn-GlyXaa designates unidentified residues.
- This amino acid sequence is homologous to some of the other low molecular weight xylanases characterized so far (e.g. the xylanase from Bacillus subtilis, for reference see Paice et al., Arch. Microbiol. 144. 1986, pp. 201-206).
- Matrix assisted lase desorption ionisation time-of-flight mass spectrometry was carried out using a ToftSpecTM mass spectrometer from VG Analytical according to the manufacturers instructions.
- Matrix assisted lase desorption ionisation time-of-flight mass spectrometry gave a mass value of 20532 Da ⁇ 0.1% for the xylanase described in ex.5 having a MW of approx. 22 kDa and a pl of approx. 9.
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Abstract
Description
Claims
Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU49446/93A AU4944693A (en) | 1992-08-26 | 1993-08-25 | New xylanases having high activity and stability at alkaline conditions and high temperatures |
EP93919031A EP0663949A1 (en) | 1992-08-26 | 1993-08-25 | New xylanases having high activity and stability at alkaline conditions and high temperatures |
JP6505799A JPH08500485A (en) | 1992-08-26 | 1993-08-25 | Novel xylanase with high activity and stability under alkaline conditions and high temperature |
BR9306980A BR9306980A (en) | 1992-08-26 | 1993-08-25 | Enzyme preparation having xylanolitic activity Process for preparing an enzyme preparation using the enzyme preparation and agent containing the same |
FI950852A FI950852A (en) | 1992-08-26 | 1995-02-24 | New xylanases very active and stable under alkaline conditions and high temperatures |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DK1055/92 | 1992-08-26 | ||
DK921055A DK105592D0 (en) | 1992-08-26 | 1992-08-26 | NEW ENZYM |
Publications (1)
Publication Number | Publication Date |
---|---|
WO1994004664A1 true WO1994004664A1 (en) | 1994-03-03 |
Family
ID=8100479
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/DK1993/000277 WO1994004664A1 (en) | 1992-08-26 | 1993-08-25 | New xylanases having high activity and stability at alkaline conditions and high temperatures |
Country Status (8)
Country | Link |
---|---|
EP (1) | EP0663949A1 (en) |
JP (1) | JPH08500485A (en) |
AU (1) | AU4944693A (en) |
BR (1) | BR9306980A (en) |
CA (1) | CA2142818A1 (en) |
DK (1) | DK105592D0 (en) |
FI (1) | FI950852A (en) |
WO (1) | WO1994004664A1 (en) |
Cited By (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995027779A1 (en) * | 1994-04-11 | 1995-10-19 | Biotech International Ltd. | Bacterial xylanase |
WO1996002632A1 (en) * | 1993-03-12 | 1996-02-01 | Showa Denko K.K. | Novel xylanase, process for producing the same, method for the treatment of pulp, and production of xylo-oligosaccharides |
EP0698667A1 (en) * | 1994-07-26 | 1996-02-28 | SOLVAY & Cie (Société Anonyme) | Xylanase, microorganisms for its production, DNA molecules, process of preparation and use thereof |
WO1996013574A1 (en) * | 1994-10-26 | 1996-05-09 | Biotech International Limited | Bacterial protein with xylanase activity |
BE1008570A3 (en) * | 1994-07-26 | 1996-06-04 | Solvay | Xylanase, micro-organisms producing same, DNA molecule, preparation methodsof said xylanase and uses thereof |
WO1997009423A1 (en) * | 1995-09-07 | 1997-03-13 | Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno | Non-starch polysaccharide hydrolysing enzymes |
WO2001030984A2 (en) * | 1999-10-27 | 2001-05-03 | Aarhus Universitet | Novel halotolerant and halophilic enzymes and the use of halotolerant and halophilic enzymes |
US6300114B1 (en) | 1994-07-29 | 2001-10-09 | Rohm Enzyme Finland Oy | Sequences of xylanase and xylanase expression vectors |
US7504120B2 (en) | 2002-06-14 | 2009-03-17 | Verenium Corporation | Xylanases, nucleic acids encoding them and methods for making and using them |
WO2010089302A1 (en) | 2009-02-06 | 2010-08-12 | University Of Chile | Protein and dna sequence encoding a cold adapted xylanase |
US8043839B2 (en) | 2006-02-14 | 2011-10-25 | Verenium Corporation | Xylanases, nucleic acids encoding them and methods for making and using them |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1991010724A1 (en) * | 1990-01-10 | 1991-07-25 | Korsnäs Ab | Preparation exhibiting enzymatic delignification activity, a method of producing the same, and applications thereof |
WO1991018976A1 (en) * | 1990-06-08 | 1991-12-12 | Novo Nordisk A/S | HEMICELLULASES PRODUCED BY $i(BACILLUS STEAROTHERMOPHILUS) |
-
1992
- 1992-08-26 DK DK921055A patent/DK105592D0/en not_active Application Discontinuation
-
1993
- 1993-08-25 AU AU49446/93A patent/AU4944693A/en not_active Abandoned
- 1993-08-25 EP EP93919031A patent/EP0663949A1/en not_active Withdrawn
- 1993-08-25 JP JP6505799A patent/JPH08500485A/en active Pending
- 1993-08-25 CA CA002142818A patent/CA2142818A1/en not_active Abandoned
- 1993-08-25 BR BR9306980A patent/BR9306980A/en not_active Application Discontinuation
- 1993-08-25 WO PCT/DK1993/000277 patent/WO1994004664A1/en not_active Application Discontinuation
-
1995
- 1995-02-24 FI FI950852A patent/FI950852A/en unknown
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1991010724A1 (en) * | 1990-01-10 | 1991-07-25 | Korsnäs Ab | Preparation exhibiting enzymatic delignification activity, a method of producing the same, and applications thereof |
WO1991018976A1 (en) * | 1990-06-08 | 1991-12-12 | Novo Nordisk A/S | HEMICELLULASES PRODUCED BY $i(BACILLUS STEAROTHERMOPHILUS) |
Non-Patent Citations (3)
Title |
---|
Agric. Biol. Chem., Volume 49, No. 11, 1985, HIROSHI HONDA et al., "Purification and Partial Characterization of Alkaline Xylanase from Escherichia Coli Carrying pCX311", figures 4 and 5. * |
Agric. Biol. Chem., Volume 51, No. 3, 1987, TETSUO HAMAMOTO et al., "Nucleotide Sequence of the Xylanase A Gene of Alkalophilic Bacillus Sp. Strain C-125", Fig. 2 around Nucleotide 150. * |
Biotechnology Letters, Volume 14, No. 11, November 1992, N. GUPTA et al., "A Thermostable Extracellular Xylanase from Alkalophilic Bacillus Sp. NG-27", figures 1-2. * |
Cited By (27)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1996002632A1 (en) * | 1993-03-12 | 1996-02-01 | Showa Denko K.K. | Novel xylanase, process for producing the same, method for the treatment of pulp, and production of xylo-oligosaccharides |
WO1995027779A1 (en) * | 1994-04-11 | 1995-10-19 | Biotech International Ltd. | Bacterial xylanase |
US6346407B1 (en) | 1994-07-26 | 2002-02-12 | Genencor International, Inc. | Xylanase, microorganisms producing it, DNA molecules, methods for preparing this xylanase and uses of the latter |
US8148104B2 (en) | 1994-07-26 | 2012-04-03 | Danisco Us Inc. | Xylanase, microorganisms producing it, DNA molecules, methods for preparing this xylanase and uses of the latter |
BE1008570A3 (en) * | 1994-07-26 | 1996-06-04 | Solvay | Xylanase, micro-organisms producing same, DNA molecule, preparation methodsof said xylanase and uses thereof |
BE1008751A3 (en) * | 1994-07-26 | 1996-07-02 | Solvay | Xylanase, the producing microorganisms, dna molecules, methods of preparation of this xylanase and uses thereof. |
EP0698667A1 (en) * | 1994-07-26 | 1996-02-28 | SOLVAY & Cie (Société Anonyme) | Xylanase, microorganisms for its production, DNA molecules, process of preparation and use thereof |
AU711105B2 (en) * | 1994-07-26 | 1999-10-07 | Genencor International, Inc. | Xylanase, microorganisms producing it, DNA molecules, methods for preparing this xylanase and uses of the latter |
US7022827B2 (en) | 1994-07-26 | 2006-04-04 | Genencor International, Inc. | Xylanase, microorganisms producing it, DNA molecules, methods for preparing this xylanase and uses of the latter |
US7638613B2 (en) | 1994-07-26 | 2009-12-29 | Genencor International, Inc. | Xylanase, microorganisms producing it, DNA molecules, methods for preparing this xylanase and uses of the latter |
US6300114B1 (en) | 1994-07-29 | 2001-10-09 | Rohm Enzyme Finland Oy | Sequences of xylanase and xylanase expression vectors |
US6506593B2 (en) | 1994-07-29 | 2003-01-14 | Rohm Enzyme Finland Oy | Production and secretion of proteins of bacterial origin in filamentous fungi |
US6667170B1 (en) | 1994-07-29 | 2003-12-23 | Röhm Enzyme Finland OY | Sequences of Xylanase and Xylanase expression vectors |
WO1996013574A1 (en) * | 1994-10-26 | 1996-05-09 | Biotech International Limited | Bacterial protein with xylanase activity |
US6200797B1 (en) | 1994-10-26 | 2001-03-13 | Biotech International Limited | Bacterial protein with xylanase activity |
US6548283B1 (en) | 1994-10-26 | 2003-04-15 | Agenix Limited | Bacterial protein with xylanase activity |
WO1997009423A1 (en) * | 1995-09-07 | 1997-03-13 | Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno | Non-starch polysaccharide hydrolysing enzymes |
WO2001030984A3 (en) * | 1999-10-27 | 2001-11-15 | Univ Aarhus | Novel halotolerant and halophilic enzymes and the use of halotolerant and halophilic enzymes |
WO2001030984A2 (en) * | 1999-10-27 | 2001-05-03 | Aarhus Universitet | Novel halotolerant and halophilic enzymes and the use of halotolerant and halophilic enzymes |
US7504120B2 (en) | 2002-06-14 | 2009-03-17 | Verenium Corporation | Xylanases, nucleic acids encoding them and methods for making and using them |
US7547534B2 (en) | 2002-06-14 | 2009-06-16 | Verenium Corporation | Methods for making a composition to treat a wood, a pulp or a paper |
US8728769B2 (en) | 2002-06-14 | 2014-05-20 | Bp Corporation North America Inc. | Xylanases, nucleic acids encoding them and methods for making and using them |
US9765319B2 (en) | 2002-06-14 | 2017-09-19 | Bp Corporation North America Inc. | Xylanases, nucleic acids encoding them and methods for making and using them |
US8043839B2 (en) | 2006-02-14 | 2011-10-25 | Verenium Corporation | Xylanases, nucleic acids encoding them and methods for making and using them |
USRE45660E1 (en) | 2006-02-14 | 2015-09-01 | Bp Corporation North America Inc. | Xylanases, nucleic acids encoding them and methods for making and using them |
WO2010089302A1 (en) | 2009-02-06 | 2010-08-12 | University Of Chile | Protein and dna sequence encoding a cold adapted xylanase |
US8679814B2 (en) | 2009-02-06 | 2014-03-25 | University Of Chile | Protein and DNA sequence encoding a cold adapted xylanase |
Also Published As
Publication number | Publication date |
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DK105592D0 (en) | 1992-08-26 |
BR9306980A (en) | 1999-01-12 |
FI950852A0 (en) | 1995-02-24 |
JPH08500485A (en) | 1996-01-23 |
EP0663949A1 (en) | 1995-07-26 |
AU4944693A (en) | 1994-03-15 |
CA2142818A1 (en) | 1994-03-03 |
FI950852A (en) | 1995-02-24 |
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