171 results about "Type III collagen" patented technology

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 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 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 discloses Dong'e donkey-hide gelatin active peptide and application thereof. Relates to the technical field of biology. The invention provides an amino acid sequence and application of the Dong'e donkey-hide gelatin active peptide. The amino acid sequence of the CCA active peptide is YYTSASGDEMVSLK, and the CCA active peptide can significantly up-regulate the I-type collagen gene expression, increase the I-type and III-type collagen content and reduce senescence markers. The peptide fragment can be combined with TGFBR2, especially TGFBR1, gene and protein expression of the TGFBR2 can be up-regulated, downstream targets of a TGF-beta-SMAD pathway can be activated, and the MMPs / TIMPs ratio can be reduced. According to the present invention, the TGF beta-SMAD pathway is activated, the MMPs / TIMPs ratio is reduced, the collagen synthesis is promoted, and the excessive decomposition of the collagen is inhibited, such that the I type collagen content and the III type collagen content are increased, and the aging degree of the photoaging dermal fibroblast is reduced;

The application discloses an anti-wrinkle composition containing recombinant collagen as well as a preparation method and application thereof, and belongs to the technical field of cosmetics and skin care products. The composition contains the following active ingredients in percentage by mass: 0.01%-20% of recombinant humanized type I collagen, 0.01%-20% of recombinant humanized type III collagen, 0.01%-10% of recombinant humanized type XVII collagen, and 0.01%-20% of space-bred desert algae extract. The three kinds of recombinant humanized collagens and the space-bred desert algae extract form a complex system, and when the total mass ratio is in a specific interval of 1.3:1-4:1, the three kinds of recombinant humanized collagens and the space-bred desert algae extract can synergistically improve the skin barrier function. Based on the synergistic mechanism of the complex system, the composition disclosed by the application can effectively promote the skin moisture retention and anti-wrinkle ability, repair the skin barrier, and is suitable for the development and application of skin care products for anti-wrinkle, skin tightening and skin aging delay.

The invention discloses a freeze-dried essence with a repairing effect and a preparation method, and belongs to the field of skin care products, and the freeze-dried essence is composed of freeze-dried powder and an independent solvent liquid. The freeze-dried powder contains 0.5-3 parts of palmitoyl oligopeptide, 3-8 parts of I-type collagen, 1-5 parts of III-type collagen, 0.1-1 part of chitosan, 1-3 parts of secondary variable-temperature fermentation liquor of betula platyphylla and 2-8 parts of a freeze-drying protective agent; the solvent liquid is prepared from 0.5 to 3 parts of hyaluronic acid and 0.05 to 0.5 part of sodium polyglutamate. The preparation method comprises the following steps: a, adding fermentation liquor after dissolving; b, adding nano liposome and a protective agent; c, quickly freezing to-55 + / -2 DEG C by liquid nitrogen, and preserving heat for 2.5-3.5 hours; d, drying for 9-11 hours under the vacuum of 15-25 Pa and at the temperature of-18 + / -2 DEG C; and e, the temperature is increased to 15 + / -2 DEG C in a stepped mode, vacuum is smaller than or equal to 20 Pa, and drying is conducted for 18- Through scientific formula design, multiple effects of efficient repairing, anti-aging and relieving are achieved.

The invention provides preparation and application of an improved recombinant III-type collagen peptide. The collagen comprises an amino acid sequence as shown in SEQ ID NO.1, and a tag sequence is fused at the N terminal and / or C terminal of the amino acid sequence as shown in SEQ ID NO.1. The recombinant collagen provided by the invention only contains a small affinity tag during expression, is convenient to purify and does not need enzyme digestion, so that the process steps are greatly simplified, the process cost is reduced, the excellent characteristics of the original recombinant collagen peptide Co8 (SEQ ID NO.1) are maintained, and the recombinant collagen still has high expression quantity, high hydrophilicity and high stability.

The invention discloses a recombinant humanized type-A type-III collagen, a preparation method thereof and a recombinant bacterium, and relates to the technical field of bioengineering. According to the preparation method, peptide fragments with 100% identical sequences of III-type collagen from human are integrated into host bacteria to obtain recombinant bacteria for efficiently synthesizing recombinant humanized A-type III-type collagen, and the recombinant humanized collagen produced by utilizing the recombinant bacteria does not contain any gene sequence of non-human collagen sources, and has the advantages of high efficiency, high yield, high yield and the like. The collagen has the characteristics of small molecular weight, high yield, high purity, water solubility and the like, and can be widely applied to the fields of skin care products, medical cosmetology, skin repair dressings and the like, so that the recombinant humanized A type III collagen, the preparation method thereof and the recombinant bacteria provided by the invention have better industrial application prospects.

The invention provides a medical-grade recombinant three-type collagen and application thereof. The recombinant humanized three-type collagen has certain cell proliferation promoting capacity and is free of cytotoxicity. The medical-grade recombinant three-type collagen provided by the invention not only has the capability of promoting cell proliferation, is free of cytotoxicity and high in cell adhesion activity, but also has a three-helix structure characterized by the collagen and is extremely high in stability, the purity can reach about 85% after the collagen is induced for 120H in a 100L fermentation tank, the difficulty of a downstream purification process is greatly reduced, and the production cost is reduced. The effect in the aspect of cell activity is obvious.

The invention provides a recombinant III-type collagen with a triple helix structure as well as a preparation method and application thereof, and relates to the field of protein engineering. According to the invention, a novel humanized III-type collagen functional fragment with biological activity is obtained through screening by selecting low-immunogenicity and high-biological activity blocks after complete sequence analysis of human III-type collagen. The functional fragments obtained by screening are repeatedly combined to obtain the novel recombinant humanized III-type collagen, and the recombinant humanized III-type collagen is a novel protein with strong biological activity and low immunogenicity, and provides a novel raw material for medical instruments and skin care products. The recombinant humanized III-type collagen sequence is efficiently expressed by adopting engineering bacteria, the expression level is high, the purification method is simple, and the recombinant humanized III-type collagen sequence is suitable for industrial production and wide popularization.

The invention belongs to the technical field of natural product extraction, and particularly relates to a vaccinium macrocarpum fruit extract-containing composition which comprises the following main components in percentage by mass: 0.04-0.3% of vaccinium macrocarpum nanoemulsion, 1-5% of vaccinium macrocarpum seed oil, 0.05-10% of III-type collagen, 1-10% of multiple hyaluronic acid, 0.5-2% of rosmarinic acid and the balance of water. The preparation method of the composition containing the vaccinium macrocarpum fruit extract comprises the following steps: step 1, selecting fresh vaccinium macrocarpum fruits, performing quick freezing, performing vacuum freeze drying until the water content is less than or equal to 5%, performing crushing, and performing sieving with a 80-mesh sieve; 2, 200 mL of a citric acid buffer solution with the pH being 4.0 is added into every 100 g of fruit powder, 0.6% of compound enzyme is added, and stirring and activating are conducted for 30 min at the temperature of 40 DEG C; step 3, adding 60% acidified ethanol according to a solid-liquid ratio of 1: 3, and performing ultrasonic extraction at 50 DEG C (200W) for 1.5 hours; according to the composition disclosed by the invention, through the innovation of extraction, purification, vectorization and freeze-drying full-chain technologies, the bottlenecks of large activity loss and low transdermal rate of a traditional process are broken through, and the synergistic effect of triple effects of oxidation resistance, wrinkle resistance and barrier repair is realized.

The invention relates to a recombinant vector for expressing a natural long-sequence recombinant III-type collagen as well as a preparation method and application of the recombinant vector, and relates to the field of proteins. The nucleotide sequence for optionally coding the recombinant III type collagen has at least one section of nucleotide sequence for coding the amino acid sequence as shown in SEQ ID NO: 1 (optionally has at least one section of nucleotide sequence as shown in any one of SEQ ID NO: 2 to SEQ ID NO: 4). The engineering bacterium for efficiently expressing the III type collagen natural long sequence is constructed through a genetic engineering technology, large-scale production of the engineering bacterium is achieved, and the engineering bacterium can be widely applied to the fields of medicine, clinical skin repair, tissue engineering, beauty and the like.

The invention discloses a recombinant III-type collagen and PDRN compound composition and a preparation method thereof, and relates to the technical field of biological medicines. The composition is formed by compounding recombinant human III type collagen and polydeoxyribonucleotide according to the mass ratio of 1: 10-10: 1, the weight-average molecular weight of the recombinant human III type collagen and the weight-average molecular weight of the polydeoxyribonucleotide are 50-100 kDa, the ratio can generate synergistic interaction, fibroblast proliferation, migration and collagen secretion are remarkably promoted, and the activity of the composition is superior to that of a single component and commercially available products. The composition can be used for preparing medical instruments, cosmetics or water-light injections for skin repair, rejuvenation or skin improvement. The invention also provides a water-light injection containing the compound composition, and the obtained preparation is stable, clear, transparent, safe and non-irritant.

The invention belongs to the technical field of biomedical materials, and particularly relates to nucleic acid for synthesizing human III-type collagen as well as an in-vivo expression method and application of the nucleic acid. The nucleic acid sequence of the synthesized human III type collagen is as shown in SEQ ID NO. 1. The invention provides an effective mRNA (messenger ribonucleic acid) nucleic acid, the nucleic acid encodes a human full-length III-type collagen gene sequence, can effectively realize expression in a human body, and is guided to translate into full-length III-type collagen through the nucleic acid in human cytoplasm and has a correctly folded space structure triple-helix collagen, namely human III-type collagen.

The invention belongs to the technical field of gene engineering, and provides a high-solubility and high-transdermal-property type III collagen as well as a preparation method and application thereof, and the amino acid sequence of the type III collagen is as shown in SEQ ID NO.1; and the N end of the III-type collagen is connected with a cell-penetrating peptide. The collagen provided by the invention has high dissolvability and high transdermal property, and also has relatively good tightening and moisturizing properties.

This invention relates to the use of recombinant type III humanized collagen and compositions comprising thereof in the ocular field. The invention provides the use of recombinant type III humanized collagen in the preparation of products for the prevention and / or treatment of ocular inflammation, and compositions comprising recombinant type III humanized collagen, and the use of these compositions in the preparation of products for the prevention and / or treatment of ocular inflammation and / or dry eye. Recombinant type III humanized collagen and compositions comprising thereof can significantly promote the renewal and repair of corneal epithelial cells, while inhibiting the release of inflammatory mediators, strengthening the ocular tissue barrier, enhancing tear film stability, and relieving ocular inflammation and dry eye.

The invention provides a collagen nano dispersion preparation as well as a preparation method and application thereof. The preparation method of the collagen nano dispersion preparation comprises the following steps: S1, dissolving type I collagen and type III collagen in an acidic aqueous medium to prepare a collagen solution; s2, regulating and controlling the temperature, the pH value and the ionic strength of the collagen solution, so that the collagen solution is self-assembled; and S3, when the average particle size of the collagen fibers reaches a target range, performing dynamic freezing on the self-assembly system. The preparation and stable dispersion of the nano-scale collagen fiber can be realized without chemical crosslinking, the uniformity, biocompatibility and safety of the product are remarkably improved, and the nano-scale collagen fiber has excellent water-light effects and the like, and can be widely applied to the field of medical cosmetology.

This invention discloses recombinant type III collagen, its preparation method, and its applications. The amino acid sequence of the recombinant type III collagen is shown in SEQ ID NO: 9. The recombinant type III collagen integrates the RGDV highly active motif and the EKEK hydrophilic stable domain with the core structure of type III collagen to achieve an integrated design of "high activity and long-lasting effect". The in vitro cell migration and collagen secretion effects are improved by more than 50% compared with wild type. Through anti-enzyme cleavage site modification and hydrophilic modification, the in vivo residence half-life is significantly prolonged to meet the long-term repair needs of the dermis.

The invention provides a method for plant transient expression of collagen and application of the plant transient expression of the collagen, and fragment collagen C3S1, C3S2, C3S3, C3S4 and C3S5 is formed by combining and connecting amino acid sequences shown as SEQ1, SEQ2, SEQ3, SEQ4 and SEQ5 respectively. According to the fragment collagen, plant tobacco can be used as a bioreactor, an expression vector is designed, the expression effect of the fragment collagen in the plant tobacco is compared, and meanwhile, a plant tobacco transient expression system is established, so that efficient expression of the fragment collagen in the tobacco is realized. By adopting the preparation method, tobacco can be utilized to express fragment collagen recombinant protein, a relatively high expression quantity is obtained, and compared with transformation by using full-length added intron or expression by using I-type collagen, the method realizes efficient expression of fragment collagen in III-type collagen in plant bioreactor tobacco, the period is short, and the yield is high. Maintenance of a cell line and a bioreactor is not needed, the production cost is remarkably reduced, and the safety index is high.

The application provides a preparation method of a composite biological scaffold for promoting anterior cruciate ligament repair and a product thereof, and the preparation method comprises the following steps: (1) selecting left-handed polylactic acid or polylactic acid-glycolic acid copolymer with a molecular weight of 250-500,000 to prepare a polylactic acid solution with a concentration of 2-5%, and then performing vacuum defoaming, sealing, slow pre-freezing and freeze-drying to obtain a polylactic acid sponge scaffold; (2) selecting type I or type III collagen with a molecular weight of 50-300,000 to prepare a collagen solution with a concentration of 1-3%, adjusting the pH value of the collagen solution to 6-8, preparing a ruthenium-peroxy disulfate solution, and mixing the collagen solution and the ruthenium-peroxy disulfate solution to obtain a mixed solution; (3) soaking the polylactic acid sponge scaffold in the mixed solution in step (2) under vacuum conditions; and (4) taking out the polylactic acid sponge scaffold, performing in-situ crosslinking through visible light irradiation, and obtaining a composite biological scaffold.

The invention relates to the technical field of biological agents, and provides a recombinant III-type collagen / platycladus orientalis essential oil nanoemulsion which is prepared from the following raw materials in percentage by weight: 15-35% of recombinant III-type collagen, 1-10% of platycladus orientalis essential oil and the balance of pure water. The invention also provides a preparation method of the nanoemulsion, which comprises the following steps: weighing the recombinant III collagen and the pure water according to the weight percentage, mixing and dissolving, then adding the platycladus orientalis essential oil, stirring and uniformly mixing to form primary emulsion, and carrying out high-speed shearing for 3-9 minutes by using a high-speed shearing emulsifying machine at the rotating speed of 8000r / min, so as to obtain the recombinant III collagen nanoemulsion. The recombinant III type collagen / platycladus orientalis essential oil nanoemulsion is obtained. The prepared oriental arborvitae essential oil nanoemulsion is uniform in size, high in stability and easy to store. The recombinant III type collagen has good biological activity and degradability, and can effectively reduce the irritation. The prepared nanoemulsion can significantly improve the antibacterial property of the platycladus orientalis essential oil nanoemulsion, and has a good prospect in application to bacterial infectious wounds.

The invention specifically discloses a fermentation process suitable for III-type collagen expression, and relates to the technical field of biological fermentation of recombinant genetic engineering bacteria. According to the method disclosed by the invention, a double-carbon-source mixed feeding strategy of alanine and methanol is used, so that the problems that when pichia pastoris expresses exogenous target protein by taking methanol as a unique carbon source and energy source, cell growth and protein expression fight for the carbon source and the energy source together, the metabolic burden of cells is increased, and the expression efficiency is low are solved; through a three-step methanol fed-batch strategy, the expression efficiency is improved, and the toxic action of methanol on cells is reduced, so that the yield of the III-type collagen is further improved. The fermentation process provided by the invention solves the problem of reducing the production cost while improving the high-efficiency and high-quality expression of the target product under the condition of high-density thallus fermentation.

The present invention provides an injectable antibacterial hydrogel that can be used for irregular wound repair. The antibacterial hydrogel is composed of modified recombinant humanized type III collagen and chitosan grafted with dihydrocaffeic acid. The modified recombinant type III humanized collagen hydrogel has a three-dimensional network structure, and the chitosan grafted with dihydrocaffeic acid is evenly distributed within the three-dimensional network structure of the modified recombinant type III humanized collagen hydrogel. The modified recombinant type III humanized collagen is formed by photocuring recombinant type III humanized collagen through photoinduced cross-linking. The present invention also provides a method for preparing the antibacterial hydrogel and its use in wound repair. This invention improves the material's ability to promote wound healing while meeting its antibacterial properties, while also enhancing its suitability for irregular wounds.

The present application relates to the field of biotechnology, in particular to the application of Radix Rabdosiae Rubescens derived exosomes in the preparation of a drug for promoting wound healing, wherein the prepared Radix Rabdosiae Rubescens derived exosomes are uniform in size, good in appearance, can promote cell proliferation, have good effects of promoting fibroblast migration and reducing scar formation; and a scar-free wound healing hydrogel containing Radix Rabdosiae Rubescens derived exosomes is also provided, wherein the hydrogel can encapsulate rEVs or ORI, can effectively increase the proportion of type III collagen in the new skin tissue, effectively reduce the formation of scars, effectively promote the healing of deep second-degree burn wounds, and effectively inhibit the MMT process of macrophages in the wound healing process, and has a wide application prospect in the field of scar prevention and treatment.

The invention discloses an application of hsa-miR-17-3p in the prevention and / or treatment of hepatic fibrosis. Through clinical sample sequencing, it is found for the first time that expression of hsa-miR-17-3p (SEQ ID NO.1) in hepatitis B related hepatic fibrosis tissue is remarkably reduced. Furthermore, a dual luciferase reporter gene experiment proves that the hsa-miR-17-3p can be directly combined with a 3'untranslated region of the TGFBR1 in a targeting manner to inhibit mRNA and protein expression of the TGFBR1, so that activation of hepatic stellate cells is effectively inhibited, and secretion of I-type and III-type collagen is remarkably reduced. The discovery not only reveals a new mechanism of hepatic fibrosis, but also provides a theoretical basis and candidate molecules for developing novel targeted therapeutic drugs based on miRNA, and has important clinical application value and wide prospects in the aspects of early intervention, treatment and diagnosis of hepatic fibrosis.

This invention discloses recombinant humanized type III collagen, its preparation method, and its applications, belonging to the field of medical materials technology. The amino acid sequence of this recombinant humanized type III collagen is any one of SEQ ID No. 2-6. Compared to traditional extraction methods, the amino acid sequence of this recombinant protein is highly consistent with that of natural human proteins, completely avoiding the biosafety risks of pathogens, prions, etc., that may exist during animal-derived extraction, while also exhibiting lower immunogenicity. Furthermore, this protein effectively maintains the natural biological activity and stability of collagen. When used as a wound dressing, this material can efficiently promote cell migration, spreading, and proliferation, accelerate tissue regeneration and wound healing, while possessing excellent biocompatibility, controllable degradation, and low immunogenicity.

The invention discloses an application of an engineered mRNA (messenger Ribonucleic Acid) medicine for coding COL3A1 protein in resisting skin photoaging. According to the method, accurate rational design and optimization are carried out on the 5'untranslated region (UTR) of the hCOL3A1 mRNA, so that the translation efficiency and the protein yield of the hCOL3A1 mRNA are greatly improved, the expression level of the hCOL3A1 target protein far beyond the conventional sequence in vivo and in vitro is realized, and the type III collagen is efficiently and endogenously synthesized in skin cells. Experimental results show that the engineered mRNA can effectively reduce the skin oxidative stress level, inhibit cell aging and promote endogenous synthesis and deposition of corium layer collagen, so that the barrier function of photoaged skin is remarkably improved, the dermis thickness is increased, the tissue structure is repaired, and systemic toxicity or immune side effects are not observed. The invention provides a novel strategy with high efficiency and safety for resisting skin photoaging.

The invention belongs to the technical field of recombinant collagen sequence analysis, and discloses a recombinant collagen sequence analysis method based on AI deep learning. Comprising the following steps: extracting a natural III-type collagen sequence; applying the natural III-type collagen sequence to the trained model to generate a thermodynamic diagram; generating an optimized natural III type collagen sequence based on the thermodynamic diagram; performing fine tuning on the AlphaFold2 model by using the fine tuning data set, and obtaining an integrated structure model by introducing spiral torsion constraint and hydroxyproline simulation parameters; loading the optimized natural III-type collagen sequence, and predicting the stability of the triple helix structure of the sequence through the structure model; and extracting a natural III-type collagen sequence which passes the stability detection, and outputting a report file by using a skin permeability prediction model to realize recombinant collagen sequence analysis based on AI deep learning.

This invention belongs to the field of biomedical materials and skin / hair repair technology, specifically a recombinant collagen freeze-dried composition targeting hair follicle functional sites, its preparation method, and its application. The composition includes recombinant human collagen, selected from recombinant type III collagen containing a Wnt-binding domain, recombinant type XVII collagen containing an NC16A domain, or a combination thereof; a freeze-drying protectant composed of trehalose and mannitol in a mass ratio of 1:2 to 2:1; and a buffer system, a phosphate buffer with a pH of 6.5-7.5 and a concentration of 5-20 mM. This invention provides precise hair follicle targeting and efficacy far exceeding that of ordinary collagen. It is effective for androgenetic alopecia, telogen effluvium, and diffuse thinning hair loss. It can be used for individuals with scalp micro-damage and sensitive scalps, simultaneously improving the scalp microenvironment and preventing hair loss from the root. Furthermore, its long-term stable effects have been verified over multiple time points.