80 results about "Beta-Mannanase" patented technology

The invention relates to the genetic engineering field, especially to a strain Bispora sp.MEY-1 which produces an acidophilic beta-mannanase and the acidophilic beta-mannanase MAN5A got from the strain having an amino acid sequence shown as SEQ ID NO.1 or 2, a gene man5A for coding said beta-mannanase having a nucleotide sequence shown as SEQ ID NO.1 or 2, and a recombinant vector containing said gene and applications. The beta-mannanase of the invention has the following advantages: the optimum pH 1.0-1.5, the optimum temperature 65 DEG C, good pH stability and thermostability, high specific activity, good protease resistance and easy for industrial fermentation production. The product of the invention can be widely used for the animal feeding-stuffs, the food, the medicament, the brewing and the energy industry as a novel enzyme preparation.

Methods and compositions of fracturing formations are provided. The fracturing fluid includes an enzyme breaker that decreases the viscosity of the fracturing fluid over time. The enzyme breaker can be used in environments having a pH value ranging from about 7 to about 12.

The invention provides a Beta-mannose which has amino acid sequences shown in SEQ ID: NO.2, SEQ ID: NO.4, SEQ ID: NO.6 and SEQ ID: NO.8. Or substitution, depletion or addition of one or multiple amino acids is carried out to the amino acid sequences shown in the SEQ ID: NO.2, SEQ ID: NO.4, SEQ ID: NO.6 or SEQ ID: NO.8 to obtain amino acid sequences of Beta-mannose with the same activity. The invention also provides a gene for coding the Beta-mannose, a recombinant vector containing the gene and a host cell containing the recombinant vector. The invention further provides a preparation method of the Beta-mannose, including the culture of the host cell provided by the invention. The Beta-mannose provided by the invention has heat-resisting property and acid resistance and a prokaryotic expression system established by the invention can be utilized for production. Furthermore, six histidine tags can be utilized for purification.

The present invention discloses a rapeseed meal enzyme-linked micro-ecological preparation and a solid fermentation method of rapeseed meal. The preparation comprises ingredients: xylanase, beta-glucanase, galactosidase, beta-mannanase, cellulase, phytase, acid protease, neutral protease, papain, alkaline protease, amylase, lactic acid bacteria, bacillus, yeasts, starch and polysaccharides. The preparation is prepared by mixing the various enzyme preparations with the fermentation micro-organisms, reduces the use of an acidifier, reduces production costs, and at the same time enhances decomposition and detoxification effects of the preparation. A produced fermented rapeseed meal feed product is rich in 3-15% of lactic acid; glucosinolate is reduced by 80% or more and toxic effects on livestock and poultry are significantly reduced; crude proteins are increased by 1-2% and a consumption of non-protein materials is ensured; a small peptide content is increased to 15-25% and utilization of animals for feed proteins is effectively improved; and bitter substances in the rapeseed meal are effectively removed and a tannin degradation rate is 60% or more.

The invention discloses a [beta]-Mannanase in feed and a preparation method thereof, belonging to the technical fields of microbial fermentation and enzyme engineering. The invention obtains, by means of mutagenesis and sieving, an Aspergillus niger MA-56 CGMCC No.2722 which can take an abundant and inexpensive agricultural byproduct as a fermentation raw material and in which the resulting acidic [beta]-Mannanase has relatively strong stability to heat and pH; and the invention proposes a production method comprising the steps of: taking the strain as a production strain to be inoculated in a solid culture medium that is formulated by bran, dregs of beans and konjaku flour based on the weight part of 60-90:10-40:1-6; fermenting, culturing, drying and detecting the strain to prepare the [beta]-Mannanase in feed. The enzyme keeps the temperature for 1 hour at 70 DEG C, which still can maintain the enzyme activity of about 85%, and keeps the temperature for 2 hours in a pH range from 3.0 to 9.0, which still can maintain the enzyme activity of over 90%, therefore, the enzyme is relatively suitable for being used as a feed additive. The [beta]-Mannanase in feed can be popularized and applied in the field of feed production.

The invention discloses a compound enzyme preparation for a hen feed as well as a production method and an application thereof. The enzyme activity composition of the compound enzyme preparation comprises 12000-15000 U / g of xylanase, 1000-1200 U / g of beta-glucanase, 500-600 U / g of cellulose, 500-750 U / g of beta-mannanase, 500-750 U / g of pectinase, 3000-4000 U / g of acid protease, 1000-1200 U / g of neutral protease, 750-1000 U / g of mesophilic alpha-amylase, 1500-2000 U / g of fungal alpha-amylase and 2000-2500 U / g of glucoamylase. The compound enzyme preparation is diluted by using a mixture of equal mass parts of corn flour and calcium sulphate; and the addition amount of the product of the invention is 150-200 g per ton of complete formula feed.

The invention provides a heat-resistant beta-mannanase and its coding gene, recombinant bacterium and use. The heat-resistant beta-mannanase as a protein is shown in the formula (a) or (b). The heat-resistant beta-mannanase shown in the formula (a) comprises the amino acid sequence 1 of the sequence table; and the heat-resistant beta-mannanase shown in the formula (b) is a protein derived from the amino acid sequence 1 though replacement and / or deletion and / or addition of multiple amino acid residues and having a beta-mannanase activity. The invention also provides the coding gene of the heat-resistant beta-mannanase and the recombinant bacterium which is engineering bacterium and contains the coding gene. The recombinant bacterium is recombinant pichia pastoris. The heat-resistant beta-mannanase has a high specific activity, good acid stability and a wide catalysis pH range, and is suitable for being used as an additive of monogastric animals such as pigs and chickens.

The invention puts forward a cloning method for complete mRNA and DNA sequences of a novel acidic beta-mannanase gene deriving from the Aspergillus niger LW-1 strain. Nucleotide sequences of the gene are respectively SEQ ID NO:1 and SEQ ID NO:2. Bioinformatics analyses show that the xylanase belongs to the 5th family of glycoside hydrolases and is named as An Man5A, with an amino acid sequence of SEQ ID NO:3 and a corresponding gene named as An man5A. The invention also discloses a method for An Man5A construction as well as high expression and purification of recombinant GS115 / man5A. The optimal temperature and pH of the prepared recombinant Man5A are respectively 70DEG C and 4.0, and the recombinant Man5A can be stable under pH of 2.5-7.5 and a temperature lower than 60DEG C, thus boasting great industrial production potential and economic value.

The invention discloses a method for rapidly preparing galactomannan oligosaccharide enzymatic hydrolysate by utilizing guar gum. The method is characterized by firstly preparing a guar gum solution with a citric acid-sodium citrate buffer solution, then controlling the temperature and time to carry out heating swelling, after cooling, adding beta-mannanase, xylanase and cellulase at the same time, fully mixing the materials uniformly and controlling the enzymolysis temperature and time to carry out degradation. The method has the effects of achieving rapid enzymolysis of guar gum at high concentration, obtaining the galactomannan oligosaccharide enzymatic hydrolysate and saving the drying and purifying costs during subsequent production of galactomannan oligosaccharide.

The present invention relates to a palm bio-feed additive, a preparation method thereof and a bio-feed containing the additive. The palm bio-feed additive comprises the following components in parts by weight: 985-995 parts of palm meal and 5-15 parts of digestive auxiliary materials. The digestive auxiliary materials comprise probiotics and compound enzymes at a mass ratio of 1:1. The probioticscomprise the following micro-organisms in parts by weight: 1-3 parts of bacillus subtilis powder, 1-3 parts of bacillus licheniformis powder and 0.5-1.5 parts of lactic acid bacterium powder. The compound enzymes comprise the following enzymes in parts by weight: 1-3 parts of xylanase, 1.5-4.5 parts of beta-mannanase, 0-1 part of acidic protease, 0-1 part of cellulase, 0-0.5 part of beta-glucanase, 0-0.5 part of medium-temperature alpha-amylase, 0-0.3 part of pectinase, 0-0.7 part of neutral protease, 0-0.5 part of alkaline protease and 0-0.5 part of alpha-galactosidase. The palm bio-feed additive prepared by using the components in the above ratios has characteristics of improving digestion and absorption of livestock and poultry, and being small in side effects and high in utilization rate of amino acids.

The invention discloses beta-mannanase and a preparation method thereof. The beta-mannanase provided in the invention is a protein which is a protein 1, a protein 2 or a protein 3, wherein the protein 1 is composed of a 22-416 amino acid sequence shown in a sequence 1 in a sequence table; the protein 2 is composed of an amino acid sequence represented by the sequence 1 in the sequence table; and the protein 3 is obtained through substituting and / or deleting and / or adding one or more amino acid residues to an amino acid residue sequence of the protein 1 or the protein 2, has activities of the beta-mannanase, and is derived from the protein 1 or the protein 2. Engineering bacteria formed by introducing protein coding gen into Pichia pastoris are fermented in a 5L fermenting tank in a high density manner, and the enzymatic activity of a fermenting solution can reach 50029.6U / mL (the protein content is 6.1mg / mL), so efficient express is realized. The beta-mannanase of the invention has application potentials in the food industry, the medicine industry, the papermaking industry, the forage industry, the petroleum exploitation industry, the fine chemical engineering industry and the like.

The invention relates to the field of genetic engineering, and specifically relates to a high-activity beta-mannanase MAN5A with a pH value within the range of 2.5-6.5, and a gene and an application thereof. The invention provides the novel beta-mannanase MAN5A with an amino acid sequence represented by SEQ ID No.1 or 2. The invention also provides a gene coding the beta-mannanase MAN5A. The nucleotide sequence of the gene is represented by SEQ ID No.3 or 4. The invention also provides a recombinant vector and a recombinant strain containing the gene, and an application thereof. The high-activity beta-mannanase MAN5A with the pH value within the range of 2.5-6 is highly acidophilic, and has a wide action pH range, good heat resistance, and good protease resistance. The beta-mannanase MAN5A can be applied in industries such as feedstuffs, foodstuffs, medicines, and the like. With a technical scheme provided by the invention, beta-mannanase which has excellent property and which is suitable for industrial application can be produced with a genetic engineering method.

The invention discloses a method for rapidly preparing galactomannan oligosaccharide enzymatic hydrolysate by utilizing a high-concentration guar gum solution. The method is characterized by firstly preparing a guar gum solution with a citric acid-sodium citrate buffer solution, then controlling the temperature and time to carry out heating swelling, after cooling, adding beta-mannanase, xylanase and endoglucanase at the same time, fully mixing the materials uniformly and controlling the enzymolysis temperature and time to carry out degradation. The method has the effects of achieving rapid enzymolysis of guar gum at high concentration, obtaining the galactomannan oligosaccharide enzymatic hydrolysate and saving the drying and purifying costs during subsequent production of galactomannan oligosaccharide.

The invention discloses a bacterial strain for producing beta-mannanase and a production process of the bacterial strain. The bacterial strain is named as bacillus subtilis-hzbzbc and preserved in the China General Microbiological Culture Collection Center (CGMCC) located at No.3 building, No. 1 courtyard, Beichen West Road, Chaoyang District, Beijing, the preservation number is CGMCC No. 4358, and the preservation time is November 22, 2010. The activity of the beta-mannanase produced by the bacterial strain is high, the optimum pH value is 2.5, the optimum temperature is 50 DEG C, and the beta-mannanase has good pH value and temperature stability. Meanwhile, the stability of the mannanase to metal ions and chelating agents is good.

The invention relates to a producing strain of Beta mannanase. The classification and nomenclature of the producing strain of Beta mannanase is Bacillus subtilis with accession number of CGMCC No.2077. The most suitable pH and temperature for the glucanase produced by the producing strain of Beta mannanase of the invention are respectively 6.0 and 70 DEG C, enzyme activity changes little under the temperature of 50-70 DEG C and the rest enzyme activity is 20 percent after the glucanase being treated for 5 hours under the temperature of 60 DEG C. The invention has fairly good heat resistance and research value.

The invention provides a feed additive, a preparation method and application thereof and relates to the technical field of livestock and poultry breeding. The feed additive is prepared from the following components: beta-glucanase (3.0-5.0)*105 U / kg, beta-mannanase (0.5-1.0)*105 U / kg, xylanase (4.4-6.6)*106 U / kg, cellulase (3.0-6.0)*104U / kg, bacillus subtilis greater than or equal to 6000 CFU / kg,bacillus licheniformis greater than or equal to 4000 CFU / kg, bacillus coagulans greater than or equal to 5000 CFU / kg, lactobacillus plantarum greater than or equal to 4000 CFU / kg, clostridium butyricum greater than or equal to 6000 CFU / kg and antimicrobial peptide of 5% or above. The feed additive can improve the utilization rate of daily ration of livestock and poultry and improve the immunity oflivestock and poultry. The use of the feed additive for livestock and poultry feed can significantly improve the production performance of livestock and poultry, reduce the cost of feed, and also reduce the use rate of antibiotics, and the additive is environmentally friendly.

The invention provides a preparation method of konjak gum serial products, belonging to the konjak gum modification technique. The characteristics of high viscosity and low solubility of the konjak gum limits the application range of the konjak gum. The method of the invention comprises the following steps: compositing beta-glucanase and beta-mannanase, controlling the enzyme proportion, enzyme addition amount and enzymolysis time to enzymatically degrade the high concentration konjak powder so as to reduce the viscosity of the konjak powder; and processing by drying and the like to obtain white, standardized serial products with an adjustable viscosity of 500-5,000mpa.s. Due to the different functions and application characteristics of degradation products with different viscosities, a product with a certain viscosity can be added into jelly, soft sweets and drinks as a food additive to play a role of retaining water, gelatinizing and thickening. The complex enzyme technique of the invention can shorten the enzymolysis time, and the controllable degradation technique expands the use range of konjak gum and has high application value.

The invention provides a method for preparing mannooligosaccharide from konjac powder. The mannooligosaccharide is obtained by utilizing the synergistic effect of recombinant beta-mannanase (reAuMan5AN3C3) and recombinant beta-endoglucanase (reAuMan5AN3C3) from a laboratory to perform enzymolysis on the konjac powder to obtain mannooligosaccharide. The catalytic efficiency of a key enzyme used by the method is relatively high, the production and using cost of the enzyme is relatively low, the period is short, the pollution is avoided, and the method has relatively great industrial production and application potential and economic values.

The invention relates to a method for producing beta-mannanase through fermentation by utilizing konjac flour microorganisms, relating to a method for producing beta-mannanase by utilizing the fermentation of konjac flour. in the method, the problems that konjac flour is currently used as the carbon source for producing beta-mannanase so that the fermentation culture medium of microorganisms is sticky, cooling is slow after sterilization, the stirring resistance is large, and the like are solved. The fermentation method comprises the following steps: 1) preparing primary fermentation culture medium; 2) adding konjac flour, then inoculating the seed liquor of microorganisms; and 3) fermenting, and adding konjac flour again, and fermenting continously. The method is used to produce beta-mannanase.

The invention relates to bio-enzyme gel breaker and applications thereof. The bio-enzyme gel breaker is an aqueous solution of beta-mannanase, alpha-galactosidase, and a cross-linking agent. According to an application method, by volume, 80 to 120 parts of a fracturing fluid base solution is mixed with 5 to 10 parts of the bio-enzyme gel breaker, and after gel forming, an obtained mixture is taken as a fracturing fluid. According to applications, galactosidase is used for removing a part of galactose residues on the side chains of galactomannan, so that mannose glycosidic bonds on the main chain of galactomannans are exposed, and then degradation effect of mannanase is adopted for selective degradation of galactomannan. Combination of the above two enzymes is capable of realizing high-efficiency rapid gel breaking of the fracturing fluid, reducing molecular weight of gel breaking products greatly, and reducing reservoir damage.

The invention belongs to the technical field of molecular biology, and in particular, relates to a recombinant bacillus subtilis strain expressing and secreting beta-mannanase efficiently and a methodfor obtaining the strain. Specifically, the method comprises the steps: a codon-optimized beta-mannanase gene is cloned to the downstreams of different signal peptide sequences in a pMA5 vector to secrete to extracellular sites; then restriction factors involved in the process of secretion are overexpressed in a host bacteria genome; and finally promoters on the expression vector are replaced, and the production of beta-mannanase is increased at the level of protein synthesis by enhancing transcription and translation processes. The recombinant bacillus subtilis expressing and secreting beta-mannanase efficiently utilizes a 2*SR culture medium for fermentation production of mannanase under conditions of the temperature of 37 DEG C and the rotating speed of 220 rpm, the highest enzyme activity of beta-mannanase in 72 h fermentation broth can reach 2207 U / mL, and the recombinant bacillus subtilis strain is a strain with good application prospect.

The invention relates to a metagenome-derived beta-mannanase, and an encoding gene and an expression thereof. A beta-mannanase gene is cloned from a biogas slurry metagenome library by the inventor, a protein encoding the gene can well hydrolyze a glycosidic bond in an incision mode, and the gene can be highly expressed in various hosts. The beta-mannanase has good stability and high activity under neutral-weak acidic conditions, and can be well applied in industrial production.

The invention discloses a construction method of a genetic engineering strain for high expression and easy purification of beta-mannanase. The method is characterized by: designing primers according to a gene sequence published by NCBI (National Center for Biotechnology Information), and adding a histidine sequence to a downstream primer, cloning a beta-mannanase gene with a Bacillus subtilis JNA chromosome as a template, conducting double digestion to a PCR product and a plasmid pMA5 with BamH I and Mlu I, carrying out overnight connection at a temperature of 16DEG C with T4DNA ligase, transforming a competent Bacillus subtilis 168 strain so as to obtain a recombinant strain BPM1003, further purifying the centrifuged fermentation broth by an Ni-NTA column, thus obtaining purified beta-mannanase, the enzyme activity of which can reach 3768.7u / ml and the specific activity of which reaches 4579.2u / mg.

The invention discloses enterobacter ludwigii and application thereof. A bacterial strain is enterobacter ludwigii MY271 collected in China Center for Type Culture Collection (CCTCC) in March 31, 2015, and has a collection number of CCTCC M2015182. According to the bred bacterial strain disclosed by the invention, beta-mannanase can be produced with high yield by using konjaku flour, manno-oligosaccharides can also be obtained, and the obtained manno-oligosaccharides have very good biological regulation functions, can be used for effectively reducing the cholesterol level of a human body, reducing blood sugar and promoting the growth of bifidobacteria in intestinal tracts, and are good food additives.

The invention relates to a method for producing low temperature-resistance beta-mannase by using enterobacter. The enterobacter sp. N18 is inoculated to a medium containing carbon source and nitrogen source, and is cultured for 12-96 hours at 37 DEG C, wherein a broth is a beta-mannanase product. Compared with the prior art, the natural enterobacter can be naturally screened for fermentation and culture to obtain the beta-mannanase, cost is low, and operation is simple. The beta-mannanase has the advantages of high enzyme activity and good stability, activity can be kept under condition that temperature is 0-100 DEG C, pH value is 3.0-11.0. and salinity is in 0-4.5M scope, and specific activity can reach 5484U / mg at 20 DEG C. In addition, viscosity of a guar gum solution with viscosity of more than 800 mPa.s can reduced to more than 95% in 10 minutes by the enzyme.

The present compositions and methods relate to an endo-Beta-mannanase cloned from Geobacillus tepidamans, polynucleotides encoding the endo-Beta-mannanase, and methods of use thereof. Formulations containing the endo-Beta-mannanase are highly suitable for use as detergents.

The invention provides a method for preparing galacto-mannan-oligosaccharides (GMOS). The method comprises the following steps: with guar gum as a substrate, compounding and degrading with hydrogen peroxide and ascorbic acid; removing residual hydrogen peroxide in the system by utilizing catalase, and adding commercial acidic beta-mannanase for performing enzymolysis; performing enzyme deactivation on hydrolysate, centrifuging to remove impurities, concentrating, precipitating to remove carbohydrates of large molecular weight by utilizing 3-5 times ethanol, performing alcohol precipitation on supernatant, performing suspended evaporation and concentrating to the solid content of more than or equal to 20%, performing spray drying, thereby obtaining GMOS dried powder with the molecular weight of less than 10000Da; and performing wet granulation on GMOS dried powder with 80%-95% thanol, thereby obtaining solid particles with the granularity of 40-100 meshes and high solubility.

The invention discloses a high-purity konjac mannan polysaccharide. The high-purity konjac mannan polysaccharide is a polysaccharide system with a weight-average molecular weight of 6000-8000Da prepared by producing konjac powder by virtue of beta-mannanase controllable hydrolysis and purifying. The polysaccharide system has physiological functions of proliferating vaginal beneficial bacteria andinhibiting harmful bacteria and can be applied to antibacterial treatment of colpitis.

The invention discloses a biological feed additive capable of raising lean meat frequency of animals, comprising the following ingredients: 8-12wt% of beta-mannanase, 1.5-2.0wt% of soya bean oil, 1.8-2.5wt% of lysine, 0.04-0.07wt% of anti-oxidant, and the balance of corn protein powder. The biological feed additive capable of raising lean meat frequency of animals is prepared by firstly preparing beta-mannanase, and then mixing beta-mannanase with soya bean oil, lysine, anti-oxidant and corn protein powder in proportion. The biological feed additive can be used as an additive of a corn soybean meal daily ration with the additional amount of 1-3%. The additive disclosed herein is a pure biological product with safety and no toxicity, can raise the absorption and conversion rate of feed, significantly improve the body shape of pigs, effectively raise the lean meat frequency of animals, and reduce body fat contents.