41 results about "Endocellulases" patented technology

The present invention relates to a heterologous exo-endo cellulase fusion construct, which encodes a fusion protein having cellulolytic activity comprising a catalytic domain derived from a fungal exo-cellobiohydrolase and a catalytic domain derived from an endoglucanase. The invention also relates to vectors and fungal host cells comprising the heterologous exo-endo cellulase fusion construct as well as methods for producing a cellulase fusion protein and enzymatic cellulase compositions.

A Clostridium thermocellum thermostable cellulase enzyme with both endocellulase activity and exocellulase activity that is able to degrade cellulose in the absence of scaffolding and other cellulosomic proteins is provided. The use of the enzyme to degrade cellulosic materials to soluble sugars is also provided.

The invention provides a feeding complex ferulic acid esterase additive, which comprises four enzymes of ferulic acid esterase, cellulase, xylanase and glucanase; among which, the enzyme activity of endo cellulose is more than 16U / g, the enzyme activity of xylanase is more than 1024U / g, the enzyme activity of beta-glucanase is more than 2.0U / g, and the enzyme activity of ferulic acid esterase is more than 2.0U / g. The feeding complex ferulic acid esterase additive of the invention has the advantages of remarkably improving the release amount of reducing sugar, the cell wall degrading rate and the vitro rumen fermentation volatile fatty acids yield in the hydrolysis process of forage plants, crop straws and bran feed, enhancing the feed biological transfer rate in an all-round way. The invention has a high practical application value in the feed industry and the ruminant breeding industry.

The invention discloses a trichoderma reesei recombinant strain capable of highly producing cellulase and application thereof. Vlbltr which grows an incompatible repression protein gene is transformed into trichoderma reesei Rut-C30, so as to obtain the trichoderma reesei Vib1 (CCMCC No.13578). Compared with the strain Vib1 and the trichoderma reesei Rut-C30, the trichoderma reesei recombinant strain has the advantages that the excretion of extracellular protein and the production of cellulase are obviously improved; compared with the trichoderma reesei Rut-C30, the activity of cellulase in the Vib1 in cellulose and bran culture mediums is increased by about 200%, and reaches 3.3U / ml; the enzyme activities of cellobiohydrolase, endo-cellulase and beta-glucosidase are respectively improved; the excretion amount of the extracellular protein in the Vib1 strain is equal to 2.19 times of the excretion amount of trichoderma reesei Rut-30; proved by the results of hydrolysis reaction of pretreated corn straw, compared with the Rut-30 strain, the output of glucose in the Vib1 strain cellulase hydrolyte is increased by 20%.

Disclosed herein are cellulase compositions useful as papermaking performance additives for improving paper dry strength of a paper product and reducing refining energy in papermaking processes, and improving paper production. These cellulase compositions are formulated using cellulase, papermaking contaminant control polymers, protein stabilizers and cellulase enhancers. These cellulase compositions measure higher in endo-cellulase activity with better stability than conventional cellulase, and have shown differentiating performance in improving paper dry strength properties versus cellulase alone.

Disclosed herein are cellulase compositions useful as papermaking performance additives for improving paper dry strength of a paper product and reducing refining energy in papermaking processes, and improving paper production. These cellulase compositions are formulated using cellulase, papermaking contaminant control polymers, protein stabilizers and cellulase enhancers. These cellulase compositions measure higher in endo-cellulase activity with better stability than conventional cellulase, and have shown differentiating performance in improving paper dry strength properties versus cellulase alone.

The invention relates to a small molecular cellulose endocellulase gene, wherein the gene is shown by the nucleotide sequence of SEQ ID NO. 1 in a sequence table; Or a sequence having 90% or more homology with the nucleotide sequence shown in SEQ ID NO. 1 and encoding the same biological function protein; Or a sequence capable of hybridizing to a nucleotide sequence shown in SEQ ID NO: 1 and encoding a protein having the same biological function. As that nucleotide sequence of SEQ ID NO. 1 in the sequence table, the pET32 vector and the Escherichia coli expression strain can be use, the fusionof the expressed product and solubilizer Trx is realized, a large amount of soluble small molecular cellulose endocellulase fusion proteins were obtained and purified by simple affinity chromatography to obtain high purity and high activity recombinant small molecular cellulose endocellulase, which can improve the yield and quality of cellulase and accelerate the production and popularization ofcellulase.

The invention relates to a cellulose endocellulase gene, wherein the nucleotide sequence of the gene is shown as SEQ ID NO. 1, and the protein encoded is shown as SEQ ID NO. 2. The invention also provides a method for efficiently expressing and purifying the recombinant protein, the purity of the purified protein is as high as 95%, and the enzyme activity of hydrolyzing cellulose to produce glucose is provided, which has important industrial value for the production of biofuel ethanol, food, feed and other industries.

The present invention relates to a heterologous exo-endo cellulase fusion construct, which encodes a fusion protein having cellulolytic activity comprising a catalytic domain derived from a fungal exo-cellobiohydrolase and a catalytic domain derived from an endoglucanase. The invention also relates to vectors and fungal host cells comprising the heterologous exo-endo cellulase fusion construct as well as methods for producing a cellulase fusion protein and enzymatic cellulase compositions.

The invention discloses a novel enzyme preparation for improving the separation efficiency of starch and protein in the processing of corn starch. The preparation is characterized by comprising ingredients such as endocellulase, exocellulase, endoxylanase, exoxylanase, pectinase and mannose. The preparation can quickly and efficiently degrade non-starch polysaccharides in corn slurry, so that the viscosity of the slurry is reduced, and the separation efficiency of the starch and the protein in corn kernels can be remarkably improved; therefore, the yield of the corn starch and corn protein powder is increased; and the washing and drying efficiencies of fiber can be improved, the utilization rate of equipment can be improved, and the energy consumption can be reduced.

The invention discloses a gene of endo-cellulase derived from Paenibacilluspolymyxa as well as preparation and an application of the endo-cellulase. The gene of the endo-cellulase is cloned on an escherichia coli expression vector according to the technical method of genetic engineering to obtain an escherichia coli recombination strain capable of heterologously expressing the endo-cellulase, andthe endo-cellulase prepared through the heterologous expression of the strain effectively degrades konjac glucomannan (also known as konjac glucomannan). The provided endo-cellulase can be widely applied to the fields of preparation of feed additives, medicines and oligo-glucomannan in agriculture and foods.

The present invention discloses one kind of clostridium and its culture process and application. The clostridium is rumen clostridium H1 in the preservation number of CGMCCNo.2125, and has high culture efficiency and contained cellulose degrading composite enzyme system comprising exocellulase, endo cellulase, xylanase, mannanase, esterase and pectase. The rumen clostridium H1 has capacity of decomposing natural cellulose, esterase activity of decomposing ester, easy medium temperature culture at 39 deg.c and capacity of utilizing pentose. It may find its wide application in cellulose degrading industry.

The present invention discloses one kind of clostridium and its culture process and application. The clostridium is rumen clostridium H1 in the preservation number of CGMCCNo.2125, and has high culture efficiency and contained cellulose degrading composite enzyme system comprising exocellulase, endo cellulase, xylanase, mannanase, esterase and pectase. The rumen clostridium H1 has capacity of decomposing natural cellulose, esterase activity of decomposing ester, easy medium temperature culture at 39 deg.c and capacity of utilizing pentose. It may find its wide application in cellulose degrading industry.

The invention provides aspergillus oryzae fermentation liquor and a preparation method of the aspergillus oryzae fermentation liquor. The aspergillus oryzae fermentation liquor contains manganese peroxide with the enzymatic activity being 100-10000U / L, endo-cellulase with the enzymatic activity being 150-3000U / L, beta-dextranase with enzymatic activity being 150-3000U / L, and beta-mannase with enzymatic activity being 50-2000U / L. The invention further provides a process technology of corn stalk sugar liquor. The corn stalk sugar liquor uses corn stalks as main raw materials and is prepared by adding a hydrogen dioxide solution to degrade the corn stalks through the aspergillus oryzae fermentation liquor. The process technology has the advantages of being short in production cycle, small in occupied space, low in production cost and high in sugar yield. In addition, the prepared corn stalk sugar liquor can be used as a carbon source raw material for producing butanol, ethyl alcohol and other fermentation industries.

The invention concerns an enzymatic method for making oenological tannins starting with lumps of wood and an enzymatic method for transforming tannins into tannins for wine-making purposes. The inventive method comprises a step which consists in contacting the lumps of wood or tannins with an aqueous solution comprising a composition containing enzymes of the cellulase class. The invention also concerns an enzymatic composition mainly consisting of enzyme of the cellulase class for making oenological tannins, comprising an endocellulase activity, a xylanase activity a beta-mannanase and / or alpha-amylase activity.

This invention is about preparation and application of beta-1,4- gluglucosan-6,2,3-sulfuric ester. This preparation is as followings: add cellulose into ion liquid, stir to form the homogeneous system of cellulose and ion, then add sulfation agent, stir at 30-35deg C and 800-1000r / min for 6-8hous. Add the alcohol-alkali liquor to precipitate the product, filter, wash to get raw esterification product. Dissolve it in the NaH2 (PO4)3-Na2H (PO4)3 buffer, PH 7.5. Add EGI to degenerate. We finally get beta-1,4- gluglucosan-6,2,3-sulfuric ester after hyperfiltration grading, condensation with depression and vacuum drying. It could be used as cosmetic additives.

The invention discloses a complex enzyme preparation for a waste paper making technology. The complex enzyme preparation comprises the following components in percent by mass: 25-35% of neutral endopectate lyase (enzyme activity being higher than or equal to 20000 u / ml), 20-30% of endocellulase (enzyme activity being higher than or equal to 5000 u / ml), 15-25% of xylanase (enzyme activity being higher than or equal to 2000 u / ml), 10-20% of lipase (enzyme activity being higher than or equal to 200 u / ml), 5-10% of laccase (enzyme activity being higher than or equal to 20 u / ml) and 15-20% of a stabilizer, wherein the neutral endopectate lyase is prepared through fermentation of bacillus cereus WENA703; the bacillus cereus WENA703 is collected in China Center For Type Culture Collection, with the collection number of CCTCC NO: M2017368.

The invention belongs to the technical field of gene engineering, in particular to a new salt resistant endo cellulase and a coding gene thereof. The invention discloses a sequence and a coding area of the gene; the sequence of the gene is shown as SEQ ID NO: 1; the full length of the sequence is 1053bp; and 350 amino acids are coded. Escherichia coli DH5 Alpha / S664 / 6p / cel8h containing a gene plasmid is preserved in China Center for Type Culture Collection (CCTCC); and the serial number of preservation is CCTCC NO:M208233. Biology validates and shows that the endo cellulase has strong tolerance to high saline salinity. The endo cellulase can be widely applied in high saline catalysis, cellulose fermentation and other industrial fields.

The invention discloses a continuous endo-cellulase as well as a coding gene and application thereof, and belongs to the technical field of bioengineering. The nucleotide sequence of the gene is shown as SEQ ID NO.3. The amino acid sequence of the continuous endo-cellulase coded by the gene is shown as SEQ ID NO.4. The continuous endo-cellulase SmCel5A is a multifunctional enzyme of a single structural domain, the molecular weight of the continuous endo-cellulase SmCel5A is less than 40 kD, heterologous expression is easy to conduct, active protein with the activity of endo-cellulase and excision-cellulase enzyme is obtained, and the problems that the growth cycle of sporocytophaga is long, enzyme components are complex, and the cellulase is not easy to directly separate and purify are effectively solved. The enzyme has wide application potential in the industries of biological energy, feed and the like.

A preparation method for neutral esterase, and a stickies control enzyme reagent used for a waste paper papermaking process are disclosed. The stickies control enzyme reagent used for the waste paperpapermaking process includes, by weight, the following components: 50%-60% of the neutral esterase (enzyme activity>=10000 u / ml), 10%-15% of endoglueanase (enzyme activity>=5000 u / ml), 15%-25% of lipase (enzyme activity>=200 u / ml), and 20% of a stabilizer; ad wherein, the neutral esterase is prepared from fermentation of bacillus cereus. A stickies control enzyme provided by the invention can effectively hydrolyze vinyl acetate (PVAc) to make the vinyl acetate become a water-soluble low molecular substance. A hydrolyzed product can be stably dispersed on a fiber surface, and the surface viscosity is lowered, so that the possibility of agglomerating into large stickies particles is greatly reduced, and the particles can be attached to the fiber surface and taken out of a system with paper without causing quality losses of the paper, thereby enabling a phenomenon that stickies affects production to be effectively controlled. The invention also includes the preparation method of the neutral esterase.

The invention discloses a method for extracting high-purity fucoidan from brown algae, which is a method for extracting fucoidan from economic brown algae serving as a raw material by combining complex enzyme assistance and hot water extraction. By limiting the addition amount of the compound enzyme, namely adding 61.5-75.06 FPU of filter paper enzyme activity, 81-115.64 CMCU of endo-cellulase and 750.8-1117.34 CBU of cellobiase into every 100g of brown algae powder and limiting specific parameters of an extraction process, the protection of a brown algae cell wall structure on fucoidan can be effectively broken through, and under the condition that protease and pectinase are not added and auxiliary means such as microwaves and ultrasonic waves are not used, the purity of the extracted fucosan sulfate can reach 87.13%, and the content of sulfate groups is 20.25%.

The invention relates to a persistent endo-cellulase mutant with remarkably improved enzyme activity and application thereof. The mutant comprises 70th and / or 235th amino acid mutation of an amino acid sequence, and the amino acid sequence of the endo-cellulase is as shown in SEQ ID NO: 1. A single-point mutation technology is adopted, based on homologous modeling and molecular docking methods, key amino acids in an enzyme activity framework are selected to optimize the molecular structure of the persistent endo-cellulase, and the enzyme activity is improved. Compared with a wild type enzyme, the content of reducing sugar generated by degrading filter paper by the continuous endo-cellulase mutant is greatly increased, and the continuous endo-cellulase mutant has important significance on efficient degradation of a cellulose substrate and reduction of the production cost.

The invention belongs to the field of enzyme engineering, and particularly relates to recombinant cellulase with a primary product cellotetrose and a building method and application thereof. A targetside, which adjusts and controls mutation of the primary product cellotetrose, of persistent endo-cellulase CcCe19A is one or more of H123, H372, R374 and Y415. The endo-cellulase can be fast expressed in escherichia coli, and is high in enzyme activity and stability so that the endo-cellulase and other enzymes can be compounded for cellulase hydrolysis. The cellotetrose generated by means of a cellulase substrate is high in concentration and purity, and can be used as a standard substance for scientific research.

The invention provides aspergillus oryzae fermentation liquor and a preparation method of the aspergillus oryzae fermentation liquor. The aspergillus oryzae fermentation liquor contains manganese peroxide with the enzymatic activity being 100-10000U / L, endo-cellulase with the enzymatic activity being 150-3000U / L, beta-dextranase with enzymatic activity being 150-3000U / L, and beta-mannase with enzymatic activity being 50-2000U / L. The invention further provides a process technology of corn stalk sugar liquor. The corn stalk sugar liquor uses corn stalks as main raw materials and is prepared by adding a hydrogen dioxide solution to degrade the corn stalks through the aspergillus oryzae fermentation liquor. The process technology has the advantages of being short in production cycle, small in occupied space, low in production cost and high in sugar yield. In addition, the prepared corn stalk sugar liquor can be used as a carbon source raw material for producing butanol, ethyl alcohol and other fermentation industries.

The invention belongs to the technical field of gene engineering, in particular to a new salt resistant endo cellulase and a coding gene thereof. The invention discloses a sequence and a coding area of the gene; the sequence of the gene is shown as SEQ ID NO: 1; the full length of the sequence is 1053bp; and 350 amino acids are coded. Escherichia coli DH5 Alpha / S664 / 6p / cel8h containing a gene plasmid is preserved in China Center for Type Culture Collection (CCTCC); and the serial number of preservation is CCTCC NO:M208233. Biology validates and shows that the endo cellulase has strong tolerance to high saline salinity. The endo cellulase can be widely applied in high saline catalysis, cellulose fermentation and other industrial fields.

The present invention relates to two-phase liquid-solid fermentation process of producing high activity cellulase for fodder, and belongs to the field of bioengineering technology. Koningic engineering strain with high antimetabolite repression, high cellulase yield and complete endocellulose, exocellulose and glucosidase is selected and used in two-phase liquid-solid fermentation process of stereo fermentation engineering technology to produce cellulase. The fermented product has activity as high as 500-700 IU / g dry leaven, and may be further prepared through serial biochemical engineering process to obtain high purity cellulase powder with activity up to 8000-10000 IU / g. The present invention is used in fodder industry to develop stalk resource while avoiding environmental pollution.

An object of the present invention is to provide a soft biomass decomposition method, a production method for a target substance from soft biomass, and an enzyme or group of enzymes for decomposing soft biomass. Provided is a soft biomass decomposition method, including a step of bringing an enzyme selected from specific exocellulase, endocellulase, and processive endocellulase into contact with soft biomass such as bagasse and rice straw. Also provided is a production method for a target substance from soft biomass by incorporating the soft biomass decomposition method as a step. Further provided is an enzyme or group of enzymes for decomposing soft biomass selected from specific exocellulase, endocellulase, and processive endocellulase.

This invention is about preparation and application of beta-1,4- gluglucosan-6,2,3-sulfuric ester. This preparation is as followings: add cellulose into ion liquid, stir to form the homogeneous system of cellulose and ion, then add sulfation agent, stir at 30-35deg C and 800-1000r / min for 6-8hous. Add the alcohol-alkali liquor to precipitate the product, filter, wash to get raw esterification product. Dissolve it in the NaH2 (PO4)3-Na2H (PO4)3 buffer, PH 7.5. Add EGI to degenerate. We finally get beta-1,4- gluglucosan-6,2,3-sulfuric ester after hyperfiltration grading, condensation with depression and vacuum drying. It could be used as cosmetic additives.