3574results about "Glycosylases" patented technology

The invention belongs to the technical field of gene engineering, and particularly relates to a beta-glycosidase SS-BGL mutant for preparing rare ginsenosides and application of the beta-glycosidase SS-BGL mutant. The beta-glycosidase SS-BGL mutant is obtained by conducting single-site mutation or double-site mutation on beta-glycosidase SS-BGL with the amino acid sequence shown as SEQ ID NO.1. The beta-glycosidase SS-BGL mutant is obtained by conducting single-site mutation or double-site mutation on beta-glycosidase SS-BGL with the amino acid sequence shown as SEQ ID NO.2. Wherein the single site mutation is that phenylalanine at the 229 site of SEQ ID NO. 1 is mutated into proline; the double-site mutation is as follows: phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into proline, and glycine at the 301 site is mutated into proline; or the phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into proline, and the leucine at the 337 site is mutated into valine; or the phenylalanine at the 229 site in SEQ ID NO. 1 is mutated into the proline, and the glycine at the 338 site is mutated into the proline. On the basis of natural beta-glycosidase SS-BGL, the molecular structure of the beta-glycosidase SS-BGL is modified through site-directed mutagenesis, and four beta-glycosidase SS-BGL mutants with the saponin conversion activity improved are obtained.

The present disclosure provides proteins useful for preventing, treating, inhibiting and / or eliminating infestation and infection of plants caused by various phytopathogenic pests, as well as formulations comprising such proteins, polynucleotides encoding such proteins, organisms expressing such proteins, and methods for preparing such proteins. And methods of using such proteins, formulations, polynucleotides, and organisms in agricultural and other effort areas.

The present disclosure provides, in various embodiments, stable injectable particle suspensions of therapeutic biologics and hyaluronan degrading agents that permit administration without the need for resuspension. In particular, the compositions disclosed herein are highly concentrated, low-volume injectable particle suspensions that increase bioavailability of therapeutic biologics.

The invention discloses a compound enzyme preparation for improving the absorption and utilization rate of a fertilizer and a preparation method thereof, and belongs to the technical field of enzyme preparations. The compound enzyme preparation is prepared from the following components in parts by weight: 100 parts of purified water, 6 to 8 parts of modified cellulase, 10 to 12 parts of immobilized protease, 10 to 12 parts of phosphatase, 10 to 14 parts of urease, 8 to 10 parts of nitrate reductase, 10 to 12 parts of pectinase, 8 to 10 parts of chitinase, 6 to 8 parts of xylanase, 10 to 12 parts of beta-glucosidase, 10 to 12 parts of superoxide dismutase and 4.5 to 6 parts of a microbial agent, the modified cellulase is obtained by chemically modifying cellulase; the immobilized protease is prepared by the following steps: adsorbing activated and coupled modified polylactic acid and a protease solution, crosslinking with glutaraldehyde, filtering, washing and drying. The compound enzyme preparation improves the fertilizer absorption and utilization rate of plants.

The invention relates to a gene modified cell and a method for producing fucosyllactose by using the gene modified cell, in particular to a kluyveromyces kluyveromyces gene modified cell and application thereof, and belongs to the technical field of biological gene engineering. The present invention provides a genetically modified cell comprising a recombinant nucleic acid sequence encoding a polypeptide having an [alpha]-1, 2-fucosyltransferase polypeptide ([alpha]-1, 2-fucosyltransferase) activity. The alpha-1, 2-fucosyltransferase polypeptide includes, but is not limited to, polypeptides having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% and 100% identical amino acid sequences with the amino acid sequences of SEQ ID NO: 1-25. According to the gene modified cell, the fucosyllactose synthesis capability of an original strain is improved.

The invention provides a Kitasatospora xanthophycea JY01 strain and application thereof, the strain is separated from intestinal tracts of spring silkworms, has good disease-resistant and growth-promoting effects, and is specifically expressed in that cellulase, beta-1, 3-glucanase, siderophore and auxin IAA can be secreted; pot experiments show that the biocontrol efficiency of the microbial inoculum to wheat stalk rot is 57.90%, the biocontrol efficiency of the microbial inoculum to corn stalk rot is 46.41%, and the plant height and fresh weight are both significantly increased. The microbial agent is expected to be applied to preparation of biological bacterial fertilizer for effectively preventing and controlling soil-borne fungal diseases.

The present disclosure provides, in various embodiments, stable injectable particle suspensions of therapeutic biologics and hyaluronan degrading agents that permit administration without the need for resuspension. In particular, the compositions disclosed herein are highly concentrated, low-volume injectable particle suspensions that increase bioavailability of therapeutic biologics.

The present disclosure relates to a process for preparing an upcycled material from a dry raw material with a moisture content of 0-30% w / w comprising lignocellulose and derived from spent cereal and / or pseudocereal grain and / or food waste or lignocellulosic biomass or a mixture thereof, the process comprising: a) a pre-treatment comprising the steps of: i) grinding and / or pulverizing the raw material to obtain a powder capable of passing through a sieve having a mesh size of 10-1000 μm and ii) performing autohydrolysis, followed by b) enzymatic hydrolysis of the of powder obtained from step ii), wherein the enzymatic hydrolysis comprises adding an enzyme mixture comprising at least a glucanase and a xylanase, and c) a post-treatment comprising the step of grinding and / or pulverizing the enzymatically hydrolysed mixture obtained from step b) to obtain the upcycled material in the form of a flour-like product.

Methods for stimulating plant growth and / or promoting plant health using free enzymes or recombinant microorganisms that overexpress enzymes are provided. Plant seeds coated with free enzymes or recombinant microorganisms that overexpress enzymes are also provided. Compositions comprising a fertilizer and an enzyme or a recombinant microorganism that overexpresses an enzyme are provided. Modified enzymes having ACC deaminase activity, recombinant microorganisms expressing the modified enzymes, plant seeds treated with the modified enzymes or recombinant microorganisms, and methods for stimulating plant growth and / or promoting plant health using the modified enzymes or recombinant microorganisms are also provided.

The invention disclosed concerns a fixed dose combination (FDC) of pertuzumab, trastuzumab, and, optionally, recombinant human hyaluronidase (rHuPH20), which is administered subcutaneously to patients. The final efficacy and safety data for the FeDeriCa clinical trial, United States Prescribing Information (USPI) (including home-use) methods, and primary analysis of the PHranceSCa clinical trial are disclosed and claimed.

The present invention is related to the field of protein engineering technology which increases the enzymatic activity and thermal stability of human hyaluronidase which is an enzyme that hydrolyzes hyaluronic acid; and more particularly to hyaluronidase PH20 variants or fragments thereof, which comprise one or more amino acid residue substitutions in the region corresponding to the alpha-helix region and its linker region in the amino acid sequence of wild-type PH20 of SEQ ID NO: 1 and in which one or more amino acid residues at the N-terminus and / or the C-terminus are selectively cleaved additionally.Specifically, the present invention relates to PH20 variants or fragments thereof, which comprise one or more amino acid residue substitutions selected from the group consisting of T341A, T341C, T341G, S343E, M345T, K349E, L353A, L354I, N356E and I361T in wild-type PH20 having the amino acid sequence of SEQ ID NO: 1, and additionally comprise the substitution of amino acids located in the alpha-helix 8 region and / or a linker region between alpha-helix 7 and alpha-helix 8 in the amino acid sequence of wild-type PH20, and in which one or more amino acids located at the N-terminal and C-terminal regions are deleted.

The present disclosure provides proteins useful for preventing, treating, suppressing and / or eliminating infestations and infections of plants by various phytopathogenic pests, as well as formulations comprising such proteins, polynucleotides encoding such proteins, organisms expressing such proteins, and methods of using such proteins, formulations, polynucleotides and organisms in agriculture and other fields of endeavor.

The invention discloses an aspergillus niger recombinant bacterium AnCT-XynB-K-agdA and application thereof, and belongs to the technical field of microbiology and bioengineering. According to the invention, Aspergillus niger AnCat is taken as an expression host, firstly, an acid protease regulatory factor prtT is knocked out to obtain a defective strain, and then different chaperonins are subjected to fusion expression by optimizing integration sites of a laccase lcc9 expression cassette, so that the high-expression laccase chaperonin is obtained. According to the method, an expression vector pC3-5 'agdA-PcitA-XynB-L-lcc9-hph-3' agdA is constructed, a recombinant bacterium AnCT-XynB-K-agdA is further obtained, the enzyme activity of the extracellular laccase subjected to shake flask fermentation reaches 1821.2 U / L, and finally, the recombinant efficient expression of the laccase Lcc9 in aspergillus niger is realized.

The invention discloses a multifunctional cellulolytic enzyme and application thereof in degradation of agricultural by-products, and belongs to the technical field of biology. The multifunctional cellulolytic enzyme is named RuCelI, and the amino acid sequence of the multifunctional cellulolytic enzyme is as shown in SEQ ID NO. 1. Based on a rumen microbial metagenome resource mining technology, a novel multifunctional cellulose decomposition enzyme RuCelI is developed, and the enzyme has double activities of endo-cellulase and xylanase, can be used for degrading agricultural by-products, reducing the treatment cost and improving the degradation efficiency, and is beneficial to improving the utilization rate and the economic benefit of the agricultural by-products.

The invention relates to an endonuclease S2 mutant, a preparation method of the endonuclease S2 mutant and application of the endonuclease S2 mutant in antibody glycosylation transformation. The glycosidase S2 mutant comprises a carbohydrate binding module (CBM) defined by M677-D843 in SEQ ID NO: 1, and the amino acid sequence of the glycosidase S2 mutant comprises SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5.

The invention discloses a methanophilic genetically engineered bacterium for producing trehalose as well as a construction method and application of the methanophilic genetically engineered bacterium, and belongs to the technical field of industrial biology. The methanophilic genetically engineered bacterium capable of producing the trehalose is obtained by taking methanophilic bacteria methanophilic 5GB1, Alkalicicus glycogenes WONF2802 or methanophilic bacteria NG09 as host bacteria, introducing maltooligosyl trehalose synthase (MTS) into the host bacteria, and carrying out over-expression on maltooligosyl trehalose hydrolase (MTH), so as to obtain the methanophilic genetically engineered bacterium capable of producing the trehalose. Under the condition that methane or methanol serves as a carbon source, the yield of the produced trehalose can reach the gram level or above, a new low-carbon biosynthesis way can be provided for commercial production of the trehalose, and the methanophilic genetically engineered bacterium for producing the trehalose has a good industrial application prospect.

The present invention discloses an engineered Trichoderma reesei for producing lipase, and a construction method and application thereof, and belongs to the fields of molecular biology and biotechnology. The engineered Trichoderma reesei heterologously expresses a lipase having an amino acid sequence as shown in SEQ ID NO.1; in the engineered Trichoderma reesei, the genes of the original cellulases cbh1 and cbh2 and xylanases xyn1 and xyn2 in Trichoderma reesei are replaced in sequence with genes encoding the lipase; the nucleotide sequence of the gene encoding the lipase is shown in SEQ ID NO.2. The present invention successfully constructs an engineered Trichoderma reesei strain that secretes and expresses lipase. The engineered strain can be used for the production of food-grade lipase, and the produced lipase meets food safety standards, and can significantly improve the specific volume and whiteness of baked goods (such as bread), which will be more suitable for actual food industry needs.

The invention discloses a molecular modification design method for improving the catalytic activity and the thermal stability of citrobacter vinegaticus-derived hyaluronidase and expression application of the citrobacter vinegaticus-derived hyaluronidase. The interaction of citrobacter vinegaticus hyaluronidase active pocket amino acid residues and hyaluronate tetrasaccharide molecules is visually explored through Pymol, conservative analysis of pocket amino acid evolution is carried out through Consurf, and rational design is carried out through policies of FoldX for calculating enzyme-substrate binding free energy, so that the hyaluronidase catalytic activity is improved. A mutant with significantly improved enzyme activity is obtained through screening, and the thermal stability of the enzyme is further improved through PROSS design and an analysis strategy of mutation site amino acid evolution conservative property and position. According to the invention, the sequence, structure and function of hyaluronidase are deeply studied, and hyaluronidase is mutated from two aspects of stabilizing a catalytic structure and promoting a catalytic reaction, so that a high-activity enzyme with higher application value in industrial production is obtained.

The invention discloses an endoglucanase truncated mutant and an application of the endoglucanase truncated mutant. According to the invention, based on endoglucanase BsEGL from bacillus subtilis DLG, a truncated mutant is constructed by deleting a carbohydrate binding domain (CBM) of the endoglucanase BsEGL, and the truncated mutant is named as BsEGL [delta] CBM. Compared with BsEGL, the truncated mutant BsEGL delta CBM has the advantages that the thermal stability, the pH stability, the organic solvent tolerance and the like are remarkably improved, and the activity of degrading phosphoric acid expanded cellulose and corncob xylan is obtained. In addition, the enzymatic activity of the truncated mutant BsEGL delta CBM is remarkably improved by 6.41%, and the catalytic efficiency is remarkably improved. Due to the characteristics, the mutant can be suitable for industrial processes such as feed processing, papermaking, biological energy sources, spinning and food brewing, and is particularly suitable for enzyme catalytic reaction under extreme environments such as high temperature and low pH. The invention provides an enzyme preparation tool with excellent performance for efficient degradation of cellulose biomass, and has important application value.

The invention provides an H1N1 broad-spectrum mRNA (messenger ribonucleic acid) vaccine taking HA (hemagglutinin) and NA (neuraminidase) protein tandem as a target and a preparation method of the H1N1 broad-spectrum mRNA vaccine, and relates to the technical field of vaccine preparation, the amino acid sequence of the mRNA vaccine is shown as SEQ ID NO.1, and the nucleotide sequence of the mRNA vaccine is shown as SEQ ID NO.2, namely, the mRNA vaccine is obtained by connecting GGGSGGSGGGSGGGGS with conserved amino acid sequences of hemagglutinin HA and neuraminidase NA. The defects in the prior art are overcome, and the broad spectrum and the protection effect of the vaccine are improved, so that the vaccine can better cope with the immune escape of the latest variant in the future.

The invention discloses a self-assembly enzyme for directional synthesis of uniform low molecular weight dextran, the self-assembly enzyme is named as YG-SpyTag-SpyCatcher-padex, the YG is a dextran sucrase gene (Genbank No.DQ345760), the padex is a dextran enzyme gene (Genbank No.KF999646.1), the SpyTag and the SpyCatcher are two specific oligopeptides, and aspartic acid in the SpyTag and lysine in the SpyCatcher spontaneously form an isopeptide bond. The invention belongs to the technical field of biology, and relates to an enzyme engineering technology. According to the invention, dextran sucrase derived from an escherichia coli genetically engineered bacterium BL21 (DE3)-YG-SpyTag and dextran enzyme derived from a pichia pastoris genetically engineered bacterium X33-SpyCatcher-padex are combined through a SpyTag-SpyCatcher protein covalent coupling system, and the obtained self-assembled enzyme can be used for catalyzing the substrate sucrose to be directly converted into dextran with the molecular weight of 3000 to 4000 Da.

The invention belongs to the technical field of biology, in particular to the technical field of agricultural biology. More specifically, the present invention relates to methods for modulating carbon assimilate distribution to increase plant yield by gene editing (e.g., guided editing) precise introduction of environmentally responsive elements.

The invention relates to a method for constructing an enzyme polymer to improve enzyme catalytic activity. The method comprises the following steps: modifying a phage sponge protein Acb2 at a C terminal or an N terminal of glucosidase, glucuronidase, sucrose phosphorylase and the like through a genetic engineering means; the enzyme polymer with better catalytic activity is obtained by screening and optimizing the types of connecting peptides between the sponge protein and the natural enzyme and the number of amino acids. The specific enzyme activity of the sponge protein driven clustered enzyme aggregate enzyme designed and constructed in the invention is improved by 3.43 times compared with that of a wild enzyme, and kcat is improved by 32.3 times. The sponge protease polymer is high in catalytic activity, simple in preparation process and wide in application range, and has the potential of being applied to large-scale industrial production.

The invention relates to the technical field of synthetic biology, metabolic engineering and cell culture. The present invention provides a cell for the production of a negatively charged, preferably sialylated, oligosaccharide wherein the cell is genetically engineered to have or express, preferably overexpress, a hydrolytic UDP-N-acetyl-D-glucosamine-2-epimerase. The invention further provides the use of said cells in culture or incubation. Also described are methods of producing negatively charged, preferably sialylated oligosaccharides using the cells, and purification of the negatively charged, preferably sialylated oligosaccharides.

The invention belongs to the technical field of bioengineering technologies, and particularly relates to heparanase sulfate as well as a preparation method and application thereof. The amino acid sequence of the heparanase sulfate (HPSE) is as shown in SEQ ID NO. 2. The HS carbohydrate chain substrate capable of being specifically degraded by the heparan sulfate enzyme (WJS HPSE) provided by the invention is white jade snail polysaccharide (WJS PS) containing an IdoA2S-GlcNAc disaccharide repeating unit, heparin or HS. In addition, the heparan sulfate enzyme (WJS HPSE) provided by the invention is high in thermal stability and has optimal enzyme activity at the temperature of 57-67 DEG C and the pH value of 4.0, and the stability and reutilization efficiency of the heparan sulfate enzyme in a catalytic process are remarkably improved, so that the application value of the HPSE is improved.

The invention discloses methanophilic genetically engineered bacteria with high fermentable sugar yield as well as a construction method and application of the methanophilic genetically engineered bacteria, and belongs to the technical field of metabolic engineering and synthetic biology. The method comprises the following steps: by taking aerobic methanophilic bacteria, namely Methylovimicrobium burabiense 5GB1, Methylovimicrobium alcaliphilum 20Z, Alkalicicus glycogenes WONF2802 or extracellular polymer methanoigens NG09 as host bacteria, carrying out genetic modification on a sugar metabolism path in the host bacteria, and expressing a gene for producing the fermentable sugar in the host bacteria, so as to construct the methanophilic gene engineering bacteria for high yield of the fermentable sugar. The methanophilic genetically engineered bacterium produces the fermentable sugar by using methane as the sole carbon source and energy, can provide a new low-carbon biosynthesis approach for commercial production of the fermentable sugar, and has good industrial and agricultural application prospects.

Methods for stimulating plant growth and / or promoting plant health using free enzymes or recombinant microorganisms that overexpress enzymes are provided. Plant seeds coated with free enzymes or recombinant microorganisms that overexpress enzymes are also provided. Compositions comprising a fertilizer and an enzyme or a recombinant microorganism that overexpresses an enzyme are provided. Modified enzymes having ACC deaminase activity, recombinant microorganisms expressing the modified enzymes, plant seeds treated with the modified enzymes or recombinant microorganisms, and methods for stimulating plant growth and / or promoting plant health using the modified enzymes or recombinant microorganisms are also provided.

The invention discloses a high-yield beta-alanine engineering bacterium constructed based on an I-type CRISPRi screening system and a method. PckA genes encoding oxaloacetic acid to phosphoenolpyruvic acid and nadB genes encoding L-aspartic acid to oxaloacetic acid are knocked out, so that a countercurrent path for synthesizing beta-alanine is blocked, and carbon loss is reduced; the ldhA gene and the poxB gene are knocked out, so that the content of synthesized pyruvic acid is increased; through overexpression of the NCgl0580 gene, the tolerance of EcN to a product is improved; by knocking out lacI genes in a genome, the adding cost of an inducer in the fermentation process is reduced; a target gene influencing the yield of beta-alanine in engineering bacteria is screened in a high-throughput manner by optimizing an I-type CRISPRi system, so that the screening efficiency and accuracy are remarkably improved, the aspartic acid tRNA ligase gene aspS capable of improving the yield of beta-alanine is obtained, the PanD enzyme activity is increased by overexpressing the aspS, and the yield of beta-alanine is further improved.

The invention discloses application of acinetobacter baumannii bacteriophage depolymerases in preparation of bacteriostatic agents, and relates to the technical field of biology. The depolymerases disclosed by the invention can inhibit the growth of a plurality of acinetobacter baumannii, have a bacteriostatic effect on the acinetobacter baumannii, and lay a foundation for developing effective infection control strategies and treatment methods to resist infection of the multi-drug-resistant acinetobacter baumannii.