129 results about "Triple helix" patented technology

The invention provides recombinant VIII type humanized collagen and application thereof, and the recombinant VIII type humanized collagen is of a triple helix structure, has good cell adhesion activity, does not generate immunological rejection and anaphylactic reaction when being applied to a human body, is a brand new human body synthetic biological material, and has good application prospects.

The present invention belongs to the technical field of protein engineering. Provided are a recombinant type XVII collagen having a triple helix structure and the use thereof. The amino acid sequence of the recombinant type XVII collagen comprises n core units, wherein n is the integer 1 or an integer greater than 1, and the amino acid sequence of the core units is as shown in SEQ ID NO: 1. Further provided are a method for constructing a plurality of expression systems and a method for preparing the recombinant type XVII collagen. Further provided is the actual industrial application on the basis of the characteristics of the protein. The provided recombinant type XVII collagen exhibits high purity, good thermal stability and high biological activity, and has a molecular weight much smaller than that of natural collagen, thereby reducing the difficulty for the collagen to cross the skin barrier and exert biological functions. Experiments show that the recombinant type XVII collagen has a variety of biological activities such as promoting cell migration.

The invention relates to a recombinant I-type humanized collagen with crosslinking property as well as a preparation method and application thereof, and belongs to the technical field of recombinant proteins. The invention relates to recombinant I-type humanized collagen with crosslinking property. The amino acid sequence of the recombinant I-type humanized collagen comprises an amino acid sequence as shown in SEQ ID No.1 or a derivative amino acid sequence with the sequence identity of more than 90% with the sequence as shown in SEQ ID No.1; the recombinant I-type humanized collagen has a triple helix structure. A complete triple helix structure can be efficiently expressed, and the hydrogel has efficient cross-linking capability, so that the hydrogel can be efficiently and stably formed through cross-linking.

The invention discloses high-purity recombinant collagen as well as a preparation method and application thereof, the recombinant collagen is composed of a recombinant collagen monomer, and the recombinant collagen monomer has an amino acid sequence as shown in any one of SEQ ID NO: 1-22. According to the present invention, the easy-to-break site and the hydrophobicity of the collagen are optimized, such that the obtained recombinant collagen monomer and the prepared recombinant collagen have characteristics of high cell activity (adhesion, migration and proliferation), high triple helix structure stability, high purity, no cytotoxicity, large-scale production, and production cost reduction.

The present invention relates to the fields of medicine and cosmetology, and discloses a cell active peptide composition that promotes the regeneration of multi-type collagen structural proteins, its preparation method, and its application in medicine and cosmetology. The composition uses raw materials such as tripeptide-10 citrulline, hexapeptide-9, hydrolyzed sodium hyaluronate, γ-aminobutyric acid, decarboxylated carnosine hydrochloride, acetyl tetrapeptide-3, acetyl tetrapeptide-11, palmitoyl dipeptide-7, and palmitoyl pentapeptide-4. The composition can promote the coordinated expression of skin type I, type III, type IV, type VII, type XVII collagen, fibronectin, elastin, and glycosaminoglycans; prevent collagen cross-linking, maintain triple helix structure, and inhibit collagen degradation. Applied to dermatology and cosmetology, it can accelerate the regeneration of structural proteins such as skin collagen, repair skin damage, reduce wrinkles such as crow's feet, tear groove lines, nasolabial folds, and under-eye lines, solve the skin aging problem caused by the loss of structural proteins such as collagen, and improve skin elasticity and firmness.

The invention belongs to the technical field of biological medicine, and particularly relates to recombinant human XVII micromolecular collagen as well as a preparation method and application thereof. According to the invention, a recombinant human XVII type micromolecular collagen (Re-Hcol17) is constructed, and a stable triple helix structure is formed by connecting the 15th collagen domain, the 12th collagen domain, the 5th collagen domain and the 1st collagen domain in series. Experimental results prove that the Re-Hcol17 can significantly promote NIH / 3T3 fibroblast proliferation within the concentration range of 10-100 [mu] g / mL, the effect is better than that of natural human collagen, and extracellular matrix adhesion can be effectively promoted. Through genetic engineering and fermentation process innovation, the bottleneck that the human-derived XVII type collagen is difficult to efficiently express and unstable in structure is broken through, and a foundation is laid for wide application of the human-derived XVII type collagen in the fields of biological medicines and cosmetics.

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.

Provided is a method for biosynthesis of a human structural material type XVII collagen. The collagen comprises the amino acid sequence shown in SEQ ID NO: 2 or a variant amino acid sequence after mutation of the amino acid sequence. The variant amino acid sequence retains the function of the amino acid sequence as shown in SEQ ID NO: 2. The collagen can promote cell adhesion and has a triple helix structure.

The invention relates to the technical field of synthetic biology, in particular to polypeptide with a triple helix structure, recombinant XII type humanized collagen and application of the recombinant XII type humanized collagen. The recombinant XII type humanized collagen is successfully expressed and prepared, has a triple-helix structure and good biological activity including promotion of cell proliferation activity, cell adhesion activity and inhibition of MMP-1, is used as a biological material derived from a human body, does not generate immunological rejection and anaphylactic reaction when applied to the human body, and has a good application prospect. The composition can be used for medical or non-medical application of a plurality of tissues and organs of a human body, filling, compatibilizing or repairing, and promotion of skin compactness or wrinkle resistance and other scenes.

The invention belongs to the field of gene engineering. In particular to high-stability recombinant XVII type collagen as well as a preparation method and application thereof. The recombinant XVII type collagen is obtained by repeating an amino acid sequence (as shown in SEQ ID NO.1) from a natural human XVII type collagen peptide for 2-5 times. The recombinant XVII type collagen can also be secreted and expressed in a pichia pastoris expression system, the preparation method is simple, and the high-yield recombinant XVII type collagen can be obtained at low cost. The cell proliferation, migration and adhesion promoting activities of the recombinant XVII type collagen are obviously superior to those of commercially available XVII type collagen, a triple helix structure can be formed at room temperature, a 1% aqueous solution has high-temperature stability and moist heat sterilization stability, and the recombinant XVII type collagen can be widely applied to the fields of cosmetics and medical instruments.

The invention relates to the field of preparation of materials for articular cartilage repair, in particular to collagen and chitosan cross-linked temperature-sensitive gel for articular cartilage repair and a preparation method of the collagen and chitosan cross-linked temperature-sensitive gel. The cartilage repair material prepared by the invention adopts the recombinant human collagen with a triple helix structure as a main raw material. The raw material is prepared by using a mammalian cell expression system, is close to protein folding and polymerization of natural protein, and has a spatial structure and modification necessary for active protein. The recombinant human collagen with the triple-helix structure is good in biocompatibility and free of rejection reaction and anaphylactic reaction, and the risk of disease transmission possibly caused by animal-derived collagen can be avoided. The injection type temperature-sensitive gel is liquid at low temperature and is converted into hydrogel at 37 DEG C, so that the injection type temperature-sensitive gel is suitable for being used as a transfer carrier for in-vivo injection treatment, secondary injury of an organism is effectively avoided, and the injection type temperature-sensitive gel has a wide clinical application prospect.

The invention relates to a recombinant III-type human collagen expression strain with a whole chain length and a triple helix structure as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering bacteria. The recombinant pichia pastoris genetically engineered bacterium expresses recombinant III-type human collagen with a full chain length and a triple helix structure, and a ku70 gene of the recombinant pichia pastoris genetically engineered bacterium is knocked out and knocked in a recombinant 4-proline hydroxylase coding gene and an FKBP22 coding gene. The recombinant pichia pastoris genetically engineered bacterium provided by the invention can be used for obtaining the recombinant full-chain-length triple-helix recombinant III-type human collagen with a higher expression level through high-density fermentation, does not contain heterologous sequences, has higher hydroxyproline content and thermal stability, is close to natural human collagen, has a stable triple-helix structure, is high in thermal stability, and can be used for preparing the recombinant III-type human collagen. The application in the fields of medical health and medical beauty such as medical beauty filling and tissue repair can be met.

Provided in the present application is a recombinant type XVII collagen having a triple helix structure. The collagen has an amino acid sequence containing one or more amino acid residue repeating units, wherein each of the repeating units contains a sequence as shown in SEQ ID NO. 1 or SEQ ID NO. 2, and the repetition number is n, where 1＜n≤60. The recombinant type XVII collagen provided in the present application can be used in the fields of food, health care products, cosmetics, medical instruments, etc.

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 belongs to the technical field of natural active collagen, and particularly relates to a method for rapidly detecting the content of active collagen. According to the method, a collagen reference substance with a complete triple helix structure and denatured collagen losing the triple helix structure after denaturation treatment are mixed according to different proportions, a series of standard samples with different triple helix structure contents are prepared, and the content of active collagen is detected by utilizing self-assembly performance characterization. The method comprises the following steps: adding a PBS (Phosphate Buffer Solution) into a collagen solution, carrying out in-vitro self-assembly under proper conditions (cooperative control of collagen concentration, pH value, ion strength, temperature and the like), and determining the turbidity change of the collagen solution in real time under a set wavelength to obtain a quantitative standard curve for calculating the content of active collagen in a test sample. The method for detecting the content of the active collagen is quicker, simpler and more convenient, and compared with a traditional protease enzymolysis method, the detection time is greatly shortened, and the detection efficiency is also improved.

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 designs a preparation method of an electrochemical sensor for detecting acrylamide based on a Ce-MOF and AuPd nanoflower composite material. The advantages of large surface area and good stability of Ce-MOF are utilized to load a large amount of AuPd NFs, and the composite material Ce-MOF (at) AuPd NFs with high conductivity is obtained. The preparation method comprises the following steps: modifying an electrode by virtue of a physical adsorption effect, fixing a triple-helix molecular switch THMS on the electrode by virtue of a gold-sulfur bond and a palladium-sulfur bond, when acrylamide exists, combining acrylamide with an aptamer, forming DNAzyme by virtue of single-stranded CDNA1 and single-stranded CDNA2, and driving a DNA walker amplification strategy in the presence of Mg < 2 + >, so that a hairpin HP1 is sheared, and a large amount of single-stranded S1 is obtained. According to the method, S1, signal tags combined on an electrode are reduced and signals are reduced by destroying a triple helix structure, so that acrylamide detection is realized. The electrochemical biosensor constructed by the method disclosed by the invention has relatively high specificity and relatively high sensitivity.

The invention provides a composition containing recombinant collagen with a triple helix structure, and belongs to the technical field of biology. The recombinant humanized III type collagen and the recombinant humanized XVII type collagen in the composition can synergistically promote cell adhesion, promote collagen regeneration, up-regulate expression of genes related to skin hydration and down-regulate expression of inflammatory factor genes, and can be applied to products for resisting aging, tightening, moisturizing, repairing and / or relieving inflammation.

The invention relates to an expression cassette and a method for producing humanized triple helix collagen by using the expression cassette, and belongs to the technical field of genetic engineering. The expression cassette sequentially comprises a promoter sequence, a fusion gene and a terminator sequence from upstream to downstream, the fusion gene comprises nucleotide fragments A, B and C, each fragment comprises an immutable region and a variable region, and the variable region is selected from I, II, III, IV and XVII type human collagen. A kex2 protease recognition site KR is introduced into an expression box, so that an expression product can be subjected to trisection cutting, and three equal-length peptide chains with a triple-helix basic structure are formed. The invention also provides a recombinant humanized collagen with a self-assembled triple helix structure and a production method thereof, and the method comprises the following steps: preparing a recombinant expression vector containing the expression cassette and a proline hydroxylase expression vector, and transferring the two expression vectors into competent cells for co-expression. The method can be used for producing the recombinant humanized collagen with the self-assembled triple-helix structure, and has a good application prospect.

The invention specifically discloses a recombinant XVII type collagen and a preparation method thereof, and relates to the technical field of genetic engineering. The invention provides a recombinant XVII type collagen protein and a preparation method of the recombinant XVII type collagen protein. Compared with the prior art, the recombinant XVII type collagen pichia pastoris engineering strain provided by the invention is high in yield, the protein is not easy to degrade, and the recombinant XVII type collagen pichia pastoris engineering strain has a stable triple helix structure.

The invention discloses recombinant collagen III + X as well as a preparation method and application thereof, and belongs to the technical field of biochemical engineering. The amino acid sequence of the recombinant collagen III + X provided by the invention is shown as SEQ ID NO: 1, and the recombinant collagen III + X is formed by repeating human III type collagen 387-467 amino acid fragments twice and connecting human X type collagen 521-680 amino acid fragments to the C end. Experiments prove that the recombinant III + X collagen and proline hydroxylase are co-expressed to form a stable triple helix structure, and the recombinant III + X collagen has great advantages in the aspects of cell migration, safety and the like, and can be used as a raw material for preparing wound dressings and skin care products.

Methods of providing biologically synthesized human structural material Type VI collagen are provided. The collagen of the present application comprises an amino acid sequence set forth in SEQ ID NO: 2 or a variant amino acid sequence that is mutated from the amino acid sequence set forth in SEQ ID NO: 2, the variant amino acid sequence retaining the function of the amino acid sequence set forth in SEQ ID NO: 2. The collagen of the present application is capable of promoting cell adhesion and has a triple helix structure.

The invention discloses a high-biological-activity human-derived I-type collagen variant, a composite material based on the high-biological-activity human-derived I-type collagen variant and a preparation method. According to the invention, the COL109 fragment which can be efficiently secreted and expressed in pichia pastoris and has a triple helix structure is obtained through screening; an adhesion domain (GPP) 10 is further fused to improve the mechanical strength, and a variant COL109 (GPP) 10 is obtained; and finally, compounding with polycaprolactone (PCL) according to an optimized ratio (1: 1) to obtain the composite material with high biological activity (promoting osteogenic differentiation and wound healing) and excellent mechanical properties. The material is suitable for bone tissue engineering scaffolds, skin regeneration dressings and the like.

The application discloses a recombinant collagen type XVII and a preparation method thereof, and relates to the technical field of genetic engineering. The application provides a recombinant collagen type XVII. Compared with the prior art, the recombinant collagen type XVII provided by the application has high yield of Pichia pastoris engineering bacteria, is not easy to be degraded, and has a stable triple helix structure.

The invention discloses a bone repair material as well as a preparation method and application thereof. The bone repair material is prepared from recombinant human collagen, recombinant human bone morphogenetic protein 2 and hydroxyapatite, wherein the ratio of the inorganic matter to the organic matter in the bone repair material is (64-66): (36-34). The bone repair material disclosed by the invention takes the recombinant human collagen as a core raw material, so that the components and the structure of the prepared material are ensured to be highly consistent with those of natural bone tissues; secondly, by adding the recombinant human bone morphogenetic protein 2, the osteogenic activity of the material is further enhanced. The finally obtained recombinant human collagen active bone repair material completely retains the triple helix structure of the recombinant human collagen, so that the recombinant human collagen active bone repair material has complete biological activity, and also has the outstanding advantages of good osteogenic activity, no disease and virus transmission risk and no immunogenicity.

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 invention provides recombinant II-type collagen as well as a preparation method and application thereof, and belongs to the technical field of bioengineering. The recombinant II-type collagen monomer is obtained by splicing Gly-X-Y repetitive gene sequences of multiple sections of triple helix regions in human II-type collagen, immunodominant epitopes, unstable amino acid sequence 3 conjunctions and glycosylation sites are avoided during selection, then the recombinant II-type collagen monomer is repeated for multiple times, and the recombinant II-type collagen monomer is obtained. The recombinant II type collagen with the molecular weight of 40-50 kDa is obtained; the recombinant II-type collagen is beneficial to expression of pichia pastoris, has the effects of promoting adhesion and migration of cartilage cells, can also promote differentiation into cartilage cells, and has good practicability.

This invention relates to the field of biomedical materials, providing non-denatured type II collagen ultrafine powder with cartilage tissue-targeted growth and structural repair functions, its bioactivity preservation preparation method, and its application in joints. The ultrafine powder has an average particle size of 0.5-50 μm, a triple helix structure integrity rate of not less than 85%, a thermal denaturation temperature of 38-48℃, and retains chondrocyte binding domains CB10, CB11, and MMP cleavage sites. The preparation method includes raw material pretreatment, time-limited enzymatic hydrolysis, graded purification, and ultra-low temperature ball milling. The product can be used for the repair and treatment of cartilage degenerative diseases such as osteoarthritis and cartilage damage.

The invention discloses a polypeptide, a recombinant XI type humanized collagen formed by the polypeptide and application of the recombinant XI type humanized collagen. Through a large number of screening studies, it is found that a natural human XI type collagen core functional region of recombinant XI type humanized collagen with a triple helix structure can be formed in the in-vitro biosynthesis process. On the basis, the recombinant humanized polypeptide is designed and prepared on the basis of a natural XI type collagen core functional area, and the polypeptide can form a stable triple-helix structure. The recombinant XI type humanized collagen provided by the invention has good biological activities, including promotion of cell proliferation activity, promotion of cell adhesion activity and promotion of extracellular matrix generation or reconstruction, and is suitable for various medical or non-medical applications.

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.