2485results about "Connective tissue peptides" patented technology

A recombinant humanized collagen and a preparation method therefor. The recombinant humanized type III collagen can be efficiently and stably expressed in three expression systems of yeast, Escherichia coli, and CHO cells, and has the biological activity superior to that of a commercially available collagen product. The recombinant humanized type III collagen has structural characteristics and functional activity similar to those of a natural collagen, and thus has wide application prospects in biomedical materials, tissue engineering products, cosmetics, foods, health care products or drugs, medical instruments, medical cosmetology and the like.

The invention belongs to the technical field of cosmetics, and discloses anti-aging recombinant collagen as well as a preparation method and application thereof. The amino acid sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.1, and the nucleotide sequence of the anti-aging recombinant collagen is as shown in SEQ ID NO.2. The recombinant collagen CaI1 is constructed and expressed by recombining the effective sequences of the I-type collagen, the III-type collagen and the XVII-type collagen, and the recombinant collagen CaI1 is free of cytotoxicity, free of hidden danger of endotoxin and good in safety. The skin care product added with the recombinant collagen CaI1 can promote skin collagen absorption, inhibit skin aging and improve skin elasticity, and has a good application prospect.

The invention provides a recombinant IV type collagen composition for repairing a basement membrane and relieving and fading red. The inventor finds that the recombinant IV-type collagen disclosed by the patent specification inhibits VE-cadherin protein phosphorylation through a mediated PKC alpha / PYK2 / Src signal channel, protects tight connection between endothelial cells, prevents blood component leakage and reduces vascular permeability, and the recombinant IV-type collagen has the advantages that the recombinant IV-type collagen can inhibit the VE-cadherin protein phosphorylation through the mediated PKC alpha / PYK2 / Src signal channel; the dissociation of the VE-cadherin compound caused by oxidative stress can be improved, and the steady state of an endothelial barrier is maintained; therefore, the composition has good effects of repairing a basement membrane barrier, relieving and fading red. The inventor also finds that the repairing nonapeptide (GAAGLPGPK) has the effects of repairing the basal membrane barrier, relieving and fading red for the first time, and the combination of the recombinant IV type collagen and the nonapeptide in a certain proportion shows synergistic interaction in the aspects of repairing the basal membrane barrier, relieving and fading red.

A ternary deep eutectic solvent (TDES) system for a biomolecule (for example, chitin) recovery from a biomass (for example, seafood waste), and the method of preparation of ternary deep eutectic solvent (TDES) system, and a process to differentiate / extract / recover biomolecules (such as chitin) from a biomass (such as seafood waste).

The invention provides humanized recombinant XVII collagen for promoting hair follicle stem cell repair and resisting aging as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The humanized recombinant XVII type collagen comprises polypeptide derived from the XVII type collagen or repeated protein formed by N times of repeated series connection of the polypeptide serving as a unit, n is an integer from 2 to 4; the polypeptide derived from the XVII type collagen comprises a polypeptide A and / or a polypeptide B; the amino acid sequence of the A polypeptide is as shown in SEQ ID NO: 1; and the amino acid sequence of the B polypeptide is as shown in SEQ ID NO: 2. The humanized recombinant XVII type collagen not only can be massively and stably recombined and expressed, but also has cell adhesion activity, cell proliferation promotion activity, cell migration promotion activity and excellent hair follicle repair and regeneration promotion activity, and has no sensitization side effect. The invention provides a new way for preparing collagen products with rich functions.

The invention relates to a recombinant A type humanized XVII type collagen as well as construction and application thereof, the protein is the humanized XVII type collagen which is composed of an intracellular region, a transmembrane region and an extracellular region, and the amino acid sequence is as shown in SEQ No. 1. The hair follicle repairing agent has the advantages of hair follicle repairing, hair regeneration, good biological activities of cell proliferation, cell adhesion and the like, and can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering, cosmetics and the like. The gene sequence of the recombinant A type XVII collagen is 100% identical to the corresponding part of the gene sequence of the recombinant A type XVII collagen, so that the recombinant A type XVII collagen has good hydrophilicity and high activity, and has application potential in the fields of biomedical materials, beauty cosmetics, skin care products and the like.

The invention relates to recombinant III-type humanized collagen and application of a composition containing the same to eyes. The invention provides an application of recombinant III type humanized collagen in preparation of a product for preventing and / or treating eye inflammation, a composition containing the recombinant III type humanized collagen, and an application of the composition in preparation of a product for preventing and / or treating eye inflammation and / or eye dryness. The recombinant III-type humanized collagen and the composition containing the recombinant III-type humanized collagen can obviously promote updating and repairing of corneal epithelial cells, and meanwhile, can inhibit release of inflammatory mediators, enhance eye tissue barriers, enhance tear film stability and relieve eye inflammation and eye dryness.

The invention belongs to the technical field of genetic engineering, and particularly relates to recombinant collagen based on engineering bacterium expression and a preparation method thereof, and the recombinant collagen is formed by connecting repetitive units derived from human III type collagen in series for three times; the invention also discloses a base sequence of the coding gene of the recombinant collagen, efficient expression and accurate folding of target protein are realized through genetic modification of engineering bacteria, a light-operated dynamic folding process and an orthogonal purification strategy, and cascade purification of a His6-SUMO tag and an intein-CBD module is utilized to obtain the recombinant collagen. The product purity and the native conformation retention rate are remarkably improved, and the technical problems that hydroxyproline depends on exogenous addition, the folding controllability is poor and the purification efficiency is low in a traditional process are solved.

The invention belongs to the technical field of protein engineering and biology, and discloses a recombinant III-type humanized collagen, a coding gene thereof, a protein preparation, a preparation method of the protein preparation, application of the protein preparation and a skin care product. The amino acid sequence of the recombinant III type humanized collagen is as shown in SEQ ID NO. 1. According to the recombinant III type humanized collagen, the transdermal absorption efficiency is greatly improved, and meanwhile, the proliferation and migration capabilities of keratinocytes are also remarkably improved. The collagen is widely applied to the fields of medical cosmetic plastic surgery, cosmetics and the like, and has a good application prospect.

The invention provides a recombinant XVII type collagen and application thereof, and relates to the technical field of protein engineering. The novel recombinant human XVII type collagen is obtained by optimizing and screening collagen structural domain sequences of the human XVII type collagen and then combining the sequences. The recombinant human XVII type collagen is consistent with the corresponding part of the amino acid sequence of the human XVII type collagen, does not generate immunological rejection when being applied to a human body, and can be widely applied to the industries of surgical medical treatment and medical beauty. In addition, the recombinant human XVII type collagen is high in stability, and the problem of protein degradation in the process of expressing recombinant protein by pichia pastoris can be avoided. In addition, the recombinant human XVII type collagen also has the effects of high biological activity, high biological safety, promotion of cell proliferation and differentiation and promotion of hair growth, and can be applied to the field of cosmetics or medicines as a functional component.

The invention belongs to the field of biological materials, and particularly relates to recombinant V-type humanized collagen and application thereof. The invention provides a recombinant V-type humanized collagen, the amino acid sequence of the recombinant V-type humanized collagen comprises (repetitive unit) n, and the repetitive unit comprises a sequence as shown in SEQ ID NO.1; each repeating unit is directly linked and the number n of repeating units is 10. The recombinant V-type humanized collagen can comprehensively resist skin aging.

The invention relates to the technical field of biology, and particularly discloses a recombinant collagen III and application thereof in preparation of gel. The recombinant collagen III is designed by optimizing functional area sequences of human I-type and III-type collagen, the amino acid sequence is shown as SEQ ID No.2, and the recombinant collagen III has the characteristics of high stability, good hydrophilicity and low immunogenicity. The preparation method comprises the steps of expression vector construction, escherichia coli induced expression, affinity chromatography purification and renaturation. The recombinant collagen III can be prepared into a gel dressing and comprises sodium alginate, methylparaben and other components. Experiments show that the gel can effectively promote cell proliferation and has no cytotoxicity; in a mouse skin injury model, the collagen can relieve inflammation, accelerate wound healing and inhibit scar formation, and the effect of the collagen is superior to that of natural human III-type collagen. The invention provides a safe and efficient novel material for wound repair, and is suitable for medical dressings and tissue engineering.

The invention discloses a recombinant XVII type collagen engineering bacterium as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering, and the recombinant XVII type collagen engineering bacterium is realized through three steps of designing and optimizing collagen, improving translation through codon optimization and selecting a vector for cloning. By optimizing the amino acid sequence and the codon, the expression efficiency and the stability of the recombinant XVII type collagen in engineering bacteria are improved. The optimization not only improves the production efficiency of the protein, but also reduces the production cost. The recombinant XVII type collagen has potential in the medical field, especially in the fields of skin repair, wound healing, aging resistance and the like. Meanwhile, in the fields of beauty and biological materials, collagen is widely applied to products such as masks and skin care products as a basic component, and can promote skin cell regeneration and enhance skin elasticity and repair capacity.

The invention discloses humanized III-type collagen as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. According to the method, firstly, on the basis of an original humanized III-type collagen gene, cell adhesion activity is taken as an evaluation index, a better cell adhesion sequence is screened out and spliced and integrated with the original collagen gene, and brand-new humanized III-type collagen is obtained. And constructing a recombinant expression vector of the humanized III type collagen containing the adhesion sequence by taking the probiotic EcN as a host cell, and introducing the recombinant expression vector into the EcN to obtain a strain of functional recombinant probiotic. Compared with a traditional escherichia coli engineering bacterium BL21 (DE3), the recombinant probiotics have good cell adhesion activity while efficiently expressing the humanized type III collagen, the bacterial endotoxin content of the recombinant humanized type III collagen obtained through purification meets the industrial standard, and the recombinant probiotics have good application prospects. The purification cost caused by industrial removal of endotoxin can be avoided, so that the production cost is reduced, and the method has a good application prospect.

The invention relates to the technical field of synthetic biology, in particular to recombinant XVII type collagen as well as a preparation method and application thereof.The amino acid sequence of the recombinant XVII type collagen comprises a sequence shown in any one of SEQ ID NO.1-SEQ ID NO.3. The recombinant XVII type collagen can be highly expressed in an escherichia coli and pichia pastoris system, the expression system is simple, the cost is low, and the recombinant XVII type collagen is suitable for industrial production. The yield is high; the biological activity is good. The protein can be compounded with other proteins to form raw materials of tissue engineering products, cosmetics, health care products or medicines, so that the protein can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering, cosmetics and the like.

The invention discloses an intelligent gradient extraction process and method for nutritional ingredients in yak bone marrow, and belongs to the technical field of bioactive substance extraction. The extraction method comprises the following four steps: gradient pressure supercritical degreasing, double-enzyme synergistic gradient enzymolysis, acid concentration gradient demineralization and temperature gradient gelatin extraction, and through the process coupling design of gradient pressure degreasing, buffer enzymolysis, graded acidolysis and membrane separation of gelatin, the high yield of the product in the previous stage is ensured; active structures of follow-up components are reserved to the greatest extent. Meanwhile, a sensing-decision-execution closed-loop system is constructed, real-time data sources such as NIR moisture sensing, online OPA detection and XRD in-situ analysis are integrated, technological parameters are dynamically adjusted through a multi-objective optimization algorithm, and global optimization of the yield, the purity and the energy consumption is achieved.

The invention provides a collagen freeze-dried product and a preparation method thereof, and belongs to the technical field of biology. In the formula design, through the synergistic effect of the arginine (salt) and the surfactant, denaturation and aggregation of the collagen freeze-dried product in the production process and the storage period are inhibited, the stability of the product is improved, and the prepared collagen freeze-dried product is high in activity, purity and safety. In the freeze-drying process, pre-freezing parameters are optimized, and the nucleation effect of ice crystals in the freezing process of the collagen mixed solution is controlled by regulating and controlling parameters such as temperature and vacuum degree in the freeze-drying process, so that the shapes and the sizes of the crystals in the collagen freeze-dried product are more uniform, the drying time is shortened, and the production efficiency is improved. And the collagen freeze-dried product with uniform pore diameter and excellent appearance is obtained, and the uniformity and stability of batch and inter-batch products are improved.

The invention relates to a protein-based phenylboronic acid ester dynamic crosslinking hydrogel material as well as a preparation method and application thereof. The preparation method comprises the following steps: dissolving a component A, namely a phenylboronic acid modified protein polymer derivative, in a biocompatible medium to obtain a solution A; dissolving a component B, namely a macromolecular derivative containing a vicinal diol group, into a biocompatible medium to obtain a solution B; and mixing the solution A and the solution B with a certain concentration for a certain time to obtain the protein-based phenylboronic acid ester dynamic crosslinking hydrogel material. The boric acid ester dynamic cross-linking bond belongs to reversible chemical cross-linking reaction, and the prepared protein-based hydrogel material has enough mechanical strength and cheap operability. The protein-based hydrogel material disclosed by the invention can provide the effects of cell adhesion, proliferation and differentiation promotion, tissue regeneration and the like, can load and slowly release beneficial components for promoting tissue repair, is an ideal tissue engineering scaffold material, and can be applied to defect regeneration and repair of soft tissues, cartilages, bones and the like.

The invention discloses a multi-effect repair type collagen and application thereof, and belongs to the technical field of synthetic biology. Transdermal peptide TD-1, a transmembrane region of human XVII type collagen, an extracellular sixteenth non-collagen region, an extracellular fifteenth collagen region and a coding gene of a His tag are selected to be connected in series, and a gene sequence is optimized through pichia pastoris codon selection preference; and then the T-COL17R3 is constructed and expressed to obtain the T-COL17R3. Efficacy experiments prove that compared with similar products sold in the market, T-COL17R3 not only has excellent transdermal performance, but also has better cell proliferation and migration promoting capability, better capability of resisting early saccharification product ketoamine and later saccharification product dicarbonyl compounds, better capability of scavenging free radicals and resisting elastase, and can be used for preparing the anti-epidermal drug. In addition, synthesis of melanin in B16-F10 cells can be reduced, so that T-COL17R3 has excellent application potential in development of pharmaceutical compositions or skin care products with repairing, anti-wrinkle and firming or whitening and brightening effects.

The invention provides an elastin peptide-containing composition as well as a preparation method and application thereof. The elastin peptide-containing composition comprises elastin peptide, camel milk active small molecule peptide and water lily oligopeptide in a mass ratio of 1: (1-3): (1.8-2.5). The elastin peptide, the camel milk active small molecule peptide and the water lily oligopeptide which are obtained through a specific treatment mode are compounded according to a specific proportion, synthesis of aquaporin AQP3 can be promoted, the content of sodium hyaluronate is increased, and the free radical scavenging activity is improved. The composition containing the elastin peptide can quickly and efficiently improve the skin state, repair the skin, delay the skin aging, enhance the skin function and recover the elasticity. The composition containing elastin peptide can promote the proliferation and growth of fibroblasts, improve the cell viability and promote the healing of skin wounds, and is non-toxic to cells.

The invention provides a micromolecular recombinant XVII type collagen as well as a preparation method and application thereof, and relates to the technical field of protein engineering. The novel recombinant human XVII type collagen is obtained by optimizing and screening a plurality of spiral region sequences such as a collagen structural domain, a C-terminal region and a middle region of the human XVII type collagen. The amino acid composition of the recombinant XVII type collagen is consistent with that of a natural collagen alpha1 chain, immunological rejection is not generated when the recombinant XVII type collagen is applied to a human body, the recombinant XVII type collagen can be widely applied to surgical medical treatment and medical beauty industries, the molecular weight of the collagen is small, efficient secretion soluble expression in eukaryotic host cells such as pichia pastoris can be achieved, and the recombinant XVII type collagen has a good application prospect. And industrial large-scale production is carried out. According to the present invention, the verification results show that the micromolecular recombinant XVII type collagen further has basement membrane anti-aging activity, wherein the basement membrane anti-aging activity specifically comprises the improvement of the expression levels of the IV type collagen, the VII type collagen and the XVII type collagen;

The invention discloses a low-cost large-molecular-weight fish scale collagen extraction method, and belongs to the technical field of fish scale collagen. The method comprises the following steps: (1) pretreatment: repeatedly freezing and storing fish scales and unfreezing and redissolving the fish scales; 2) impurity protein removal: taking out the fish scales, and removing fish silver; soaking in a sodium chloride solution, washing and drying; 3) extracting fish scale collagen, namely extracting the dried fish scales for 1-4 hours at 40-100 DEG C according to a solid-liquid ratio of 1: (5-20) by adopting a hydrothermal extraction mode, carrying out solid-liquid separation, and collecting fish scale collagen extract liquids in different molecular weight intervals; and 4) freeze-drying the collagen extract liquid to obtain a fish scale collagen solid product. According to the method, acid or alkali solution pretreatment is not needed, and a sodium chloride salt solution is used for mildly removing impure protein. And the structure of the fish scale collagen is relatively completely maintained while the relatively high yield is ensured.

The invention relates to the technical field of biological medicine, in particular to recombinant III-type humanized collagen as well as a preparation method and application thereof. In one embodiment, the recombinant III type humanized collagen prepared by the invention has an amino acid sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 4. The recombinant III-type humanized collagen prepared by the invention has good biological activity, can promote collagen synthesis related gene expression, promote skin fibroblast proliferation, increase the content of skin collagenous fibers and elastic fibers, and improve skin elasticity and compactness; in addition, the expression of osteogenesis related genes is promoted, and the proliferation of osteoblasts is promoted. The results show that the recombinant III-type humanized collagen prepared by the invention has an extremely wide application prospect in developing related medical and cosmetic products for delaying skin aging and promoting wound repair and bone tissue healing.

The invention provides a recombinant humanized elastin with high biological activity as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering and biological materials. 100% of the amino acid sequence of the elastin is derived from human elastin and is prepared by expression of genetically engineered bacteria; the protein has a typical beta-spiral hydrophobic structural domain and a lysine-rich hydrophilic structural domain, has a good bionic structure and functional characteristics, can significantly promote proliferation, adhesion and migration of human skin fibroblasts, also has excellent antioxidant, anti-wrinkle and anti-aging activities, and can be used for preparing a biomimetic protein. The hydrogel can be widely applied to the fields of skin repair dressings, injection implants, artificial skin, biomedical materials, functional skin care products and the like; in addition, the elastin is good in solubility, high in stability, mature in expression system and suitable for large-scale production.

The invention relates to the field of gene engineering, in particular to preparation and application of recombinant collagen. Collagen mutants are obtained through site-specific mutagenesis, mutants with good effects are screened to prepare the anti-wrinkle composition, the length, area and depth of skin wrinkles can be effectively reduced, the anti-wrinkle effective rate of the collagen Q160A mutant composition is 90%, the excellent rate is 30%, and the collagen Q160A mutant composition has good application prospects in anti-wrinkle products.

The embodiment of the invention discloses human-derived recombinant III-type collagen as well as a preparation method and application thereof. The amino acid sequence of the human-derived recombinant III-type collagen comprises a basic repetitive unit, and the amino acid sequence of the basic repetitive unit is as shown in SEQ ID NO.4. The amino acid sequence of the basic repetitive unit is as shown in SEQ ID NO.3. The amino acid sequence of the basic repetitive unit is as shown in SEQ ID NO.4. The amino acid sequence of the human recombinant III type collagen is shown as SEQ ID NO.3. The invention also discloses a preparation method of the human recombinant III type collagen. According to the invention, a human recombinant III-type collagen gene with the coding molecular weight of 60kD is designed, an escherichia coli expression strain is constructed and expressed, the yield of the human recombinant III-type collagen is increased through amplification culture on a fermentation tank, and the protein purity is improved by virtue of strategies such as ultrafiltration primary purification and affinity chromatography. The human-derived recombinant III-type collagen prepared by the invention avoids potential hidden dangers such as animal diseases or viruses and immunological rejection caused by a traditional extraction method, has excellent medical performance, can effectively control the production cost and the product quality by virtue of a fermentation preparation process, and is also easy to realize large-scale production.

The invention provides a BaP4H mutant and application thereof, and belongs to the technical field of enzyme engineering, the BaP4H mutant is mutated on an amino acid sequence as shown in SEQ ID NO.1; the invention further provides a recombinant expression vector for expressing the BaP4H mutant, a recombinant cell, a preparation method and application. According to the BaP4H mutant provided by the invention, the recombinant collagen proline is efficiently hydroxylated, and the stability of the recombinant collagen is enhanced.

The present invention relates to a recombinant human type IV collagen and a preparation method therefor, which belong to the technical field of bioengineering. A recombinant human type IV collagen is designed on the basis of a partial sequence in an α2 chain of human type-IV collagen, and DNA encoding of the protein is ligated into an expression vector to construct a recombinant expression vector; then the recombinant expression vector is transformed into competent Escherichia coli cells, and a recombinant expression plasmid is extracted; after the plasmid is linearized, the linearized plasmid is transformed into an engineered strain HCPB-PPKEX2 to obtain a recombinant engineered strain; and the engineered strain is cultured, the expression of a target protein is induced, and the target protein is purified and identified. It is further demonstrated experimentally that the collagen can be completely cleaved by Kex2, Ste13 and CPB enzymes and efficiently secreted and expressed extracellularly, and the obtained sequence is completely consistent with a theoretical sequence. It is also verified experimentally that the recombinant collagen has a cell adhesion activity and a cell migration-promoting activity. The method avoids the risk of residual exogenous proteins, shortens the time and reduces the cost of the subsequent purification process, and thus has a relatively high safety and application value.

The invention discloses an active peptide capable of improving cognitive impairment as well as a preparation method and application of the active peptide. The amino acid sequence of the active peptide is shown as SEQ ID NO: 1, 2, 3, 4, 5 or 6. The invention aims to provide a novel active peptide as well as a preparation method and application thereof aiming at the problems of limited means and unstable effect of improving cognitive impairment in the prior art, the active peptide prepared by the invention can improve the problems of learning and memory impairment and cognitive function decline to a certain extent, enriches technical approaches for improving cognitive impairment, and has a wide application prospect. And a new application choice is provided for related fields.

The invention discloses a high-transdermal recombinant humanized XVII type collagen as well as a preparation method and application thereof, and belongs to the technical field of synthetic biology. According to the preparation method, transdermal peptide TD-1, different functional fragments of human XVII type collagen and coding genes of His tags are selected to be connected in series, gene sequences are optimized through pichia pastoris codon selection preference firstly, and then the high-transdermal recombinant humanized XVII type collagen is obtained; and then the recombinant humanized XVII type collagen T-COL17-NC16 and the recombinant humanized XVII type collagen T-COL17 are constructed by expression of the recombinant humanized XVII type collagen T-COL17. Effect experiments prove that compared with similar products sold in the market, the skin-care product has more excellent transdermal performance, can improve cell proliferation and migration ability, also has obvious ability of resisting early saccharification product ketoamine and late saccharification product dicarbonyl compounds, better ability of scavenging free radicals and resisting elastase, and can be used for preparing the skin-care product. Moreover, synthesis of melanin in B16-F10 cells can be reduced, and the compound is expected to become a novel functional biological raw material to be applied to the fields of pharmaceutical compositions or skin care products and the like.