287 results about "Protein engineering" patented technology

The invention provides recombinant IV type collagen and application thereof, and relates to the technical field of protein engineering. In the invention, an amino acid residue repetitive unit with good biological activity and stability in the human-derived IV-type collagen is found, and the amino acid residue repetitive unit is used as a core for repeating and constructing the recombinant IV-type collagen, so that the novel human-derived recombinant IV-type collagen with biological activity is obtained. The human-derived recombinant IV-type collagen provided by the invention is a small-molecular-weight protein, is easy to absorb by a human body, is good in safety, can remarkably promote secretion of collagen, also has the effects of promoting cell adhesion and cell migration, and has a good application prospect in the aspects of basement membrane repair and remodeling. A new functional component is provided for products for repairing and tightening the skin, resisting aging, resisting wrinkles, preserving moisture and the like.

The present invention relates to the technical field of protein engineering and provides a recombinant humanized type III collagen having a triple helix structure, a preparation method therefor, and a use thereof. The amino acid sequence of the recombinant humanized type III collagen comprises n core units, wherein n is 1 or an integer greater than 1, and the amino acid sequence of the core unit is as shown in SEQ ID NO: 1. The present invention further provides a method for constructing an expression system and a method for preparing a recombinant humanized type III collagen. The present invention further provides an actual industrial use on the basis of the characteristics of the protein. The recombinant humanized type III collagen provided by the present invention has high purity, good stability, and high biological activity. Experiments have shown that the recombinant humanized type III collagen has various biological activities such as promoting cell proliferation and migration.

The invention belongs to the technical field of gene engineering and protein engineering, and particularly relates to thermal-stable D-psicose 3-epimerase as well as a preparation method and application thereof. According to the invention, a series of D-psicose 3-epimerase DAE mutants are constructed, and six mutants which not only can retain good catalytic activity, but also have high thermal stability are obtained through screening. The catalytic temperature of the three mutants A19D / A223P / V241I is increased by 25 DEG C compared with that of a wild type, while the three mutants A19D / A223P / V241I still show the optimal thermal stability, the catalytic activity of the three mutants is improved by 17.0%, and the three mutants show good industrial suitability. The D-psicose 3-epimerase DAE mutant with improved thermal stability provided by the invention is beneficial to improving the production efficiency of the DAE in industrial application of D-psicose, and provides powerful industrial enzyme guarantee for large-scale and low-cost preparation of D-psicose.

The invention belongs to the technical direction of protein engineering in the technical field of biology, and particularly relates to a triple-helix folding and stabilizing method for recombinant collagen, which is a technology for efficiently assisting correct folding of human recombinant collagen expressed by escherichia coli and forming a stable triple-helix structure. The technology mainly solves the common key technical problems of low folding efficiency, unstable triple helix structure and the like of the recombinant collagen in an escherichia coli expression system, and obviously improves the yield of the functional recombinant collagen. The method has important application value in the fields of biomedical materials, tissue engineering, beauty and skin care and the like, and an efficient and economical technical path is provided for industrial production of recombinant collagen.

The invention relates to a chicken beta-defensin-6 mutant based on AI aided design, a recombinant expression system and application, and relates to the technical field of gene engineering and protein engineering. The chicken beta defensin-6 mutant has the following amino acids at the following five sites: lysine at the 33rd site, lysine at the 35th site, asparagine at the 49th site, arginine at the 61st site and proline at the 65th site. The chicken beta-defensin-6 mutant has high stability, activity and solubility, compared with wild chicken beta-defensin-6, the chicken beta-defensin-6 mutant has higher antibacterial activity and expression level, an efficient and safe antibacterial substitute is provided for livestock and poultry breeding, and the chicken beta-defensin-6 mutant can be widely applied to the fields of livestock and poultry breeding, animal health care and biological preservation.

The present invention relates to the field of protein engineering, and specifically relates to a Morchella esculenta peptide and the use thereof. By means of comprehensively applying bioinformatic methods such as virtual enzymolysis, activity prediction, physicochemical property and safety evaluation, and molecular docking, one Morchella esculenta peptide having the potential effects of "inhibiting oxidation + skin whitening" is screened from Morchella esculenta, which peptide has a sequence of WWVCAK. In-vitro chemical experiments verify that WWVCAK is the most effective antioxidant and tyrosinase-inhibitory peptide. Therefore, WWVCAK can exert the effects of inhibting oxidation and skin whitening via multiple targets, multiple functions and multiple pathways. WWVCAK is expected to be developed and used in cosmetics as a functional skin-care ingredient with the twin effects of "inhibiting oxidation + skin whitening".

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 / or 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 truncated 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 portion(s) of amino acids located at the N-terminal and C-terminal regions are deleted.

The invention discloses a rhamnosyltransferase mutant and application thereof, and belongs to the technical field of genetic engineering. The catalytic activity of the rhamnosyltransferase is improved through a protein engineering modification method, so that the rhamnolipid yield of the pseudomonas aeruginosa is improved, reaches 62.87 g.L <-1 > and is 1.89 times that of a wild type, and the pseudomonas aeruginosa has a good industrial application prospect.

The invention discloses recombinant XVII type collagen with an anti-inflammatory effect and a preparation method of the recombinant XVII type collagen, and belongs to the technical field of protein engineering. The recombinant XVII type collagen provided by the invention comprises a plurality of repetitive units, and the repetitive units are as shown in SEQ ID NO. 1. Compared with natural XVII type collagen, the recombinant XVII type collagen is smaller in molecular weight, more stable and capable of being directly used for product production; meanwhile, the recombinant XVII type collagen disclosed by the invention can be combined with an inflammatory factor receptor to inhibit activation of a signal channel, so that the anti-inflammatory activity is improved, and therefore, the recombinant XVII type collagen disclosed by the invention can be used as a core active component to be applied to preparation of anti-inflammatory related products, and has a relatively high development prospect.

The present application relates to the field of protein engineering, in particular to a fusion protein with an expanded PAM recognition range and application thereof. The fusion protein provided by the present application comprises: a) an amino acid sequence from a non-PI domain of xCas9; b) an amino acid sequence from a PI domain of a SpCas9 variant. The fusion protein has a more relaxed PAM restriction and significantly improved editing efficiency, and is a novel editing tool with an expanded targeting range.

The present disclosure is generally related to the fields of microbial host cells, molecular biology, protein engineering, fermentation, protein production, and the like. Certain aspects of the disclosure are related to novel promoter and 5′-untranslated region a nucleic acid (DNA) sequences.

The invention relates to a binding protein B capable of binding with klebsiella pneumoniae and application of the binding protein B, and belongs to the technical field of molecular biology and protein engineering. The binding protein B provided by the invention can be specifically bound with an extramembrane part of a specific protein P24017. OMPAKLEPN of klebsiella pneumoniae, and experiments such as bimolecular fluorescence complementation and flow cytometry prove that the binding protein B has good recognition and stable binding capacity on the klebsiella pneumoniae, can be used for identifying and screening the klebsiella pneumoniae, and has good application prospects. And a key tool is provided for targeted drug development aiming at the bacterium.

The invention belongs to the technical field of gene engineering and protein engineering, and particularly relates to a fully human antagonistic antibody taking a lactylated STAT6 protein as a target spot and application of the fully human antagonistic antibody. According to the present invention, the completely humanized antagonistic antibody using the lactylated STAT6 protein as the target spot is screened from the human scFv phage antibody library, the amino acid sequence of the completely humanized antagonistic antibody is represented by SEQ ID NO: 1, and the completely humanized antagonistic antibody has the specificity of the targeted lactylated STAT6 protein, and has the application prospect in preparation of the product for diagnosis, treatment or alleviation of sepsis immunosuppression.

The present disclosure belongs relates to a 2709 alkali protease mutant modified based on molecular dynamics calculation and use thereof in liquid detergents. The dynamic structure feature of a 2709 alkali protease derived from Bacillus licheniformis are studied by using a molecular dynamics method through computer simulation, which is a unique effective method capable of studying a relationship between the structure of the 2709 alkali protease and molecular dynamics as well as an enzyme function in an atomic level. After molecular dynamics calculation, appropriate sites are selected for protein engineering to obtain the 2709 alkali protease mutant suitable for industrial application of detergents. The protease mutant has significantly improved enzyme activity under the alkaline condition, is better in stability, not only has good decontamination effect but also has good component compatibility with components in the detergent, and can be widely applied to the field of washing industry.

This invention relates to a recombinant type V collagen, its encoding gene, and its applications, belonging to the fields of genetic engineering and protein engineering technology. Based on the amino acid sequence of the α1 chain of human type V collagen, this invention utilizes bioinformatics and other methods to design the amino acid sequence, enabling the recombinant collagen to achieve higher expression efficiency and high stability while maintaining high biological activity. A novel collagen sequence containing 178 amino acids was designed. The nucleotide sequence of this recombinant collagen was optimized using E. coli codon preference to construct a high-expression recombinant strain. The recombinant collagen produced by fermentation of this strain contains no exogenous amino acids such as tags, making it a recombinant humanized type V collagen with good safety. Compared with commercially available type V collagen products, the recombinant type V collagen prepared using the above technical solution has better antioxidant activity and barrier repair capabilities, thus possessing good practical application value.

The invention discloses a black fungus AMY protein and a coding gene and application thereof, and belongs to the technical field of protein engineering. The invention provides a black fungus AMY protein as well as an encoding gene and application thereof in order to excavate a key enzyme gene of a starch sucrose metabolic pathway in black fungus, and discovers and verifies that the black fungus AMY protein and the encoding gene thereof play a key regulation and control role in inducing synthesis of black fungus alpha-amylase through transcriptome sequencing. A solid foundation is laid for deeply exploring the function of a key gene of a starch sucrose metabolic pathway.

The application discloses an antibody glycosylation modification regulation method, device and application, relates to the technical field of biopharmaceuticals and protein engineering, and realizes directional regulation of modification types, modification sites and modification proportions of N-glycan and O-glycan of the antibody through three core processes of constructing a glycosyltransferase engineering strain, optimizing a fermentation culture system and precisely regulating modification reaction conditions; solves the technical problems of low modification efficiency, poor specificity and insufficient product uniformity in the existing modification method, can improve the target glycan modification proportion to more than 90%, the modification product purity is greater than or equal to 98%, is suitable for glycosylation modification optimization of therapeutic monoclonal antibodies, bispecific antibodies and antibody drug conjugates, significantly improves the biological activity and pharmacokinetic performance of the antibody, wherein the ADCC activity is improved by 3-5 times, and the CDC activity is improved by 2-4 times, and has the advantages of strong controllability, good adaptability to large-scale production and high cost-effectiveness.

The invention discloses a beta-glucuronidase mutant and application thereof, and belongs to the technical field of genetic engineering and protein engineering modification. The beta-glucuronidase mutant disclosed by the invention has the following mutation on the basis of an amino acid sequence as shown in SEQ ID NO.3: T512S or T512A. The catalytic efficiency of the beta-glucuronidase mutant T512S is improved by 13.92 times compared with that of AtGUS (-3t), and the beta-glucuronidase mutant T512S shows remarkable industrial application potential.

The application provides a recombinant cholesteryl esterase, a polynucleotide thereof, a recombinant plasmid, an expression system and application. The application belongs to the technical field of protein engineering, and specifically, the amino acid sequence of the recombinant cholesteryl esterase provided by the application is shown as SEQ ID No. 1, and the nucleotide sequence for encoding the recombinant cholesteryl esterase is shown as SEQ ID No. 2. The sequence of the recombinant cholesteryl esterase provided by the application has a full length of 482 amino acids, is derived from Aspergillus ruber, has the advantage of high thermal stability, and has a thermal stability deviation of less than 10% after thermal storage at 37 DEG C for 11 days and after 30 days of on-board opening, and has a wide application prospect in the field of in-vitro diagnostic reagents.

The present disclosure provides recombinant type XVII collagen having a triple helix structure and uses thereof, which belong to the field of protein engineering technology. The recombinant type XVII collagen has an amino acid sequence formed by tandem repeats of n or more core units, where n is an integer of 1 or more, and the core unit has the amino acid sequence shown in SEQ ID NO: 1. The present disclosure also provides methods for constructing multiple expression systems and methods for preparing recombinant type XVII collagen. provide.

The present invention pertains to the field of protein engineering, molecular imaging, molecular diagnostics, and biopharmaceutics. Provided herein are six unique Proteimer protein scaffolds (TEX-S2 / S3, TEX-S4, YTHDF3, PUM, DARPin, and Aca2) that demonstrate specificity and affinity comparable to antibodies across a broad range of therapeutic targets. Methods and protocols are provided for generating non-native protein aptamers, Proteimers, which are capable of binding to a diverse set of targets, including RNA, DNA, proteins, post-translational modifications, peptides, small molecules, and prosthetic groups. This platform supports the creation of biotherapeutic aptamers from Proteimer scaffolds for the treatment of various diseases.

The invention discloses a recombinant collagen, a coding gene and a preparation method and application thereof, and belongs to the technical field of gene engineering and protein engineering. On the basis of an amino acid sequence of human VII type collagen, the amino acid sequence is designed by means of bioinformatics and the like, a new recombinant collagen is constructed, further, a pichia pastoris strain for efficiently expressing the recombinant collagen is constructed by optimizing a gene sequence through codons, and the recombinant collagen is obtained. The recombinant collagen has good biocompatibility and biological activity, can remarkably promote proliferation, adhesion and migration of L929 cells, and has wide application prospects in multiple fields such as food, medicines, cosmetics or instrument products.

The invention discloses a BPS mutant HsamBPS12Mutant and application thereof, and belongs to the field of protein engineering.The BPS mutant HsamBPS12Mutant is modified into a mutant with a chalcone synthase (CHS)-like function on the basis of benzophenone synthase (BPS) in hypericum plants through precisely designed directional mutation of 12 sites, and an effective strategy is provided for modifying natural BPS to obtain new catalytic activity. The obtained mutant retains part of BPS activity, obtains CHS-like catalytic ability, has the characteristic of dual functions, and expands the application boundary of BPS enzymes in metabolic pathway engineering. The mutant can be applied to the fields of plant metabolism engineering, synthetic biology, natural product biosynthesis, efficient production of medicinal components and the like, and has good biotechnology application potential and industrial value.

The invention belongs to the technical field of protein engineering, and particularly relates to an immunoglobulin binding protein mutant with high alkali stability and application of the immunoglobulin binding protein mutant. The immunoglobulin binding protein mutant with high alkali stability provided by the invention is obtained by mutating in a Z structural domain of an amino acid sequence of staphylococcus aureus protein A, such as SEQ ID NO: 1, wherein the amino acid sequence of the Z structural domain is shown as SEQ ID NO: 1, and mutation sites comprise the 3rd amino acid, the 6th amino acid, the 9th amino acid, the 11th amino acid, the 23rd amino acid, the 25th amino acid, the 28th amino acid, the 32th amino acid, the 33rd amino acid, the 40th amino acid, the 42nd amino acid, the 43rd amino acid, the 44th amino acid, the 52nd amino acid and the 53rd amino acid. The prepared immunoglobulin binding protein mutant is prepared into affinity chromatography filler, the alkali resistance of the affinity chromatography filler and the affinity to an antibody can be remarkably improved, and the affinity chromatography filler can tolerate online cleaning of a 0.5-1.0 mol / L NaOH solution and has a good application prospect.

The invention relates to the technical field of genetic engineering and protein engineering modification, in particular to a high-temperature-resistant alkaline lipase mutant and application thereof. After the mutant containing G242I single-point mutation provided by the invention is treated at 75 DEG C for 3 minutes, the enzyme activity residual rate is up to 70.53%, the heat resistance is obviously higher than that of a wild type, efficient catalytic activity can be kept in a high-temperature alkaline environment in the industrial fields of pulping, papermaking and the like, industrial energy consumption and cost are reduced, and the application prospect is wide.

The invention belongs to the technical field of genetic engineering and protein engineering modification, and particularly relates to a high-specific-activity cellulase mutant and application thereof. Compared with wild cellulase, the specific activity of the mutant is improved by 51.9% and reaches 319 U / mg, and an unexpected technical effect is achieved. The mutant can be widely applied to the field of textile industry, is uniform in flower formation, small in flower point and strong in stereoscopic impression, most importantly, the strength loss of the fabric is very small, the strength retention rate is improved by 10-15% compared with that of the fabric treated by wild type cellulase, the use amount of the cellulase can be greatly reduced, the cost is reduced, and the application prospect is wide.

The invention relates to genetic and protein engineering and can be used in medicine and the pharmaceutical industry. Recombinant plasmid DNA pET23b-CBD-HS-ES-GLP1, pET-parB-CBD-HS-ES-GLP1 and pET23bKanR-CBD-HS-ES-GLP1 were constructed to provide the synthesis of a hybrid protein containing the GLP-1 polypeptide sequence in Escherichia coli cells. By transforming the E. coli BL21 (DE3) strain with plasmids pET23b-CBD-HS-ES-GLP1, pET-parB-CBD-HS-ES-GLP1 and pET23bKanR-CBD-HS-ES-GLP1, the CBD-HS-ES-GLP1 hybrid protein has been produced. A method of producing polypeptide GLP-1 has been developed, which involves preparing and cultivating a producer of a hybrid protein containing polypeptide GLP-1, followed by isolation and cleavage of the said hybrid protein. The invention enables the production of polypeptide GLP-1 with high yield and simplified technology.

Provided is a parent phytase variant, which relates to the technical field of protein engineering. The variant, relative to the parent phytase thereof, has one or more amino acid substitutions at positions corresponding to positions 295, 349, and 374 of SEQ ID NO: 1. Compared to the parent phytase, the variant has increased thermal stability.

The present invention relates to the field of protein engineering and therapeutics, in particular, improved protein formats and uses thereof.

This invention belongs to the field of protein engineering and biomedicine, and particularly relates to a water-soluble membrane protein, a recombinant vector, a recombinant host bacterium, and their modification methods and applications. The method involves the following steps: First, an interface mutant is constructed based on the SQTY code, and its water solubility and ligand binding ability are evaluated. If the requirements are not met, multiple low-impact transmembrane regions are screened, and after mutation modification, the interface mutant is introduced to construct a single-transmembrane combined mutant, whose water solubility and ligand binding ability are evaluated. If the requirements are still not met, the multiple low-impact transmembrane regions are combined in pairs, and the interface mutant is introduced to construct various double-transmembrane combined mutants, whose water solubility and ligand binding ability are evaluated, and the optimal double-transmembrane combined mutant is selected. This method rationally mutates CXCR4 in stages to achieve water solubility, minimizing changes to the protein's structure and other physicochemical properties, thereby maintaining or even enhancing its binding ability to the ligand CXCL12.