79 results about "Spider Proteins" patented technology

The invention provides a bio-based adhesive and a preparation method thereof. Inspired by a spider thread structure, biomass adhesive interface enhancement based on a tannic acid adhesion group is combined with construction of a differential phase separation structure by biomass-induced polylipoic acid depolymerization, and a brand new strategy of adhesion-cohesion synergistic enhancement of the adhesive strength of the adhesive is realized. According to the invention, tannic acid and carboxymethyl chitosan are introduced into a lipoic acid-based adhesive, so that the purpose of synergistically enhancing the bonding performance of the adhesive through adhesion-cohesion is achieved, and the defects of weak cohesion, low bonding strength, poor water resistance, poor moisture resistance and the like of the conventional biomass adhesive are overcome. Benefited from a new strategy of adhesion-cohesion synergistic enhancement, the biomass adhesive has good adhesive strength in various environments and materials. In addition, the biomass adhesive also has the characteristics of biological safety, antibacterial and mildew-proof properties and recoverability, so that the product has wider application and potential compared with the previous product, and has a good application prospect.

The invention belongs to the technical field of protein expression, and particularly relates to recombinant elastin with repeated amino acid sequences and a preparation method thereof.The recombinant elastin is composed of 327 AA sequence units which are repeatedly connected in series twice and a spider silk protein GAAn module, and the AA sequence units are derived from human elastin containing VPGVG peptide fragment repeating units; the invention also discloses a double-repeat tandem sequence protein obtained by serially connecting AA sequence units twice, a coding gene of the double-repeat tandem sequence protein, a recombinant elastin sequence, a coding gene of the recombinant elastin sequence, a high-expression vector and a preparation method, and the problems of low protein expression efficiency and high purification cost at present are solved.

The invention discloses a recombinant protein and application thereof in preparation of sodium alginate fibers. The recombinant protein is obtained by fusion expression of a functional protein and at least one alginic acid binding domain protein. The amino acid sequence of the alginic acid binding domain protein is as shown in at least one of SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3. The recombinant spider silk protein with the alginic acid CBM region can exert the material science characteristics in a sodium alginate-calcium ion system, the mechanical property of alginic acid fiber is greatly enhanced, and meanwhile, the biological activity of protein can be provided for an alginate fiber system.

The invention belongs to the technical field of biology, and particularly discloses a high-density fermentation method and application of recombinant spider silk protein, and the high-density fermentation method comprises the following steps: firstly, transforming an expression vector carrying spider silk protein genes into modified pichia pastoris cells to obtain a strain; the activated strain is subjected to expanding culture, high-density fermentation culture and purification, and recombinant spider silk protein is obtained; wherein the transformation means of the pichia pastoris cell is to integrate Vitreoscilla hemoglobin Vhb gene into the pichia pastoris cell by utilizing a gene recombination technology, and the expression of the Vitreoscilla hemoglobin Vhb gene is improved by carrying out metabolic engineering transformation on the pichia pastoris cell for expression and optimizing the regulation and control of staged fermentation parameters, so that the expression efficiency of the Vitreoscilla hemoglobin Vhb gene is improved. The problems that in existing recombinant protein fermentation, oxygen dependence is high, product degradation is serious, purity is low, and large-scale stability is poor are solved, the yield and purity of recombinant spider silk protein are guaranteed, and large-scale production of the recombinant spider silk protein is facilitated.

Cosmeceutical compositions comprising a major ampullate spidroin protein (MaSp)-based fiber are disclosed. Articles comprising the compositions are further disclosed.

The application discloses a kind of imitative spider silk cellulose gel fibers and its preparation method and application, method includes: using DMAc to activate cellulose powder and obtain activated cellulose powder;Activated cellulose powder is added in DMAc / LiCl solvent and dissolved, and obtain cellulose solution;Using microfluidic equipment, cellulose solution is injected into deionized water and solidified, and washing is obtained cellulose gel fiber;Cellulose gel fiber is soaked in PVA-PAA precursor liquid, and then placed under ultraviolet lamp irradiation, then freeze and thaw, and obtain cellulose / PVA / PAA hydrogel fiber;Cellulose / PVA / PAA hydrogel fiber is pre-stretched, then soaked in CuSO4 solution, and then washed with deionized water after soaking, and obtain imitative spider silk cellulose gel fiber.The imitative spider silk cellulose gel fiber provided by the application has strong toughness, flexible conductivity and environmental resistance.

Compared with the prior art, the method for extracting the spider silk fibroin has the advantage that the content of the spider silk fibroin can be remarkably increased.

A recombinant spider silk protein comprises an NT domain, a REP domain and a CT domain. The REP domain comprises a set of domains according to the formula pA1-pG-pA2. pG represents a glycine-rich domain and pA1 and pA2 represent alanine-rich domains. One of pA1 and pA2 is a poly-alanine domain and the other of pA1 and pA2 is a poly-alanine domain having every third or fourth alanine residue replaced by an isoleucine residue or a valine residue. Spider silk spun from the recombinant spider silk proteins of the invention have improved mechanical properties.

Compared with the prior art, the method for extracting the spider silk fibroin has the advantage that the content of the spider silk fibroin can be remarkably increased.

The invention belongs to the technical field of protein, and aims to solve the problem that the application range of spider silk particles as a drug carrier is limited due to the fact that the spider silk particles are in negative net charge under a neutral condition and can only load drugs with positive charges or neutral charges at present. The novel recombinant spider silk protein is prepared on the basis of an N connection sequence of araneus ventricosus MaSp1, has positive net charge under a neutral pH value, can load a negatively charged drug, and expands the application range of the spider silk protein as a drug carrier; and a drug release test result shows that the recombinant spider silk protein is superior to MaSp2-based silk protein particles in the aspect of etoposide delivery.

The invention relates to the technical field of biology, in particular to a recombinant coral-spider silk fusion protein and functional alginate fiber and application thereof. The recombinant coral-spider silk fusion protein provided by the invention comprises the recombinant coral-spider silk fusion protein formed by fusing spider silk protein or rattan silk protein and coral protein. The invention designs a fusion protein only containing spider silk protein and coral protein. The recombinant coral-spider silk fusion protein is applied to alginate fibers, colored spinning of the alginate fibers can be achieved, and the spun fibers are high in color fastness, superior to original fibers in strength and elongation and bright and stable in color.

The invention belongs to the technical field of biology, particularly discloses a method for preparing spider silk protein-containing composite silk by using transgenic silkworms and application of the spider silk protein-containing composite silk, and particularly provides a method for synthesizing and secreting spider silk protein by using a silkworm silk gland bioreactor. The spider silk protein gene is introduced into a bombyx mori genome in a virus infection manner by utilizing a transgenic bombyx mori technology and is specifically expressed in bombyx mori silk gland cells, and the spider silk protein target gene can be introduced in batches by virtue of a lentivirus transfection method, so that bombyx mori capable of synthesizing and secreting complete spider silk protein can be developed on a large scale; the spider silk protein is synthesized and secreted through bombyx mori silk gland cells and enters silkworm cocoons along with the spinning and cocooning behaviors of bombyx mori, the silk is collected, the spider silk protein-containing composite silk with the bombyx mori silk as the background is obtained, a foundation is laid for mass production of black spider silk protein, and meanwhile a foundation is laid for improving the mechanical performance of the silk.

To provide a method for purifying recombinant silk polypeptide sustainably and by an environment-friendly process.SOLUTION: A method for isolating a recombinant spider silk protein from a host cell culture solution includes: a) a step of obtaining a cell culture solution, in which the cell culture solution contains a host cell and a growth medium, and the host cell expresses a recombinant spider silk protein; b) a step of collecting a part of the cell culture solution containing the recombinant spider silk protein; c) a step of incubating a part of the cell culture solution in an aqueous solution under an alkaline condition, by which the recombinant spider silk protein is solubilized in the aqueous solution; and d) a step of isolating the recombinant spider silk protein from the aqueous solution, by which an isolated recombinant spider silk protein sample is produced.SELECTED DRAWING: Figure 1

The invention provides a cement-based material with a spider silk-like microstructure and a preparation method of the cement-based material, and belongs to the technical field of cement-based materials and preparation of the cement-based materials. The cement-based material comprises the following raw materials: double-component cement, reinforced fibers, mineral whiskers and an additive, the two-component cement comprises silicate cement and sulphoaluminate cement. The expansion effect generated by the hydration reaction of anhydrous calcium sulphoaluminate in the double-component cement not only counteracts the partial shrinkage effect generated by the hydration of Portland cement clinker minerals, but also extrudes the reinforced fibers, so that the binding force between the fibers and a cement matrix is increased. Besides, a secondary frame formed by the generated ettringite raphioid crystals and the added mineral whiskers is combined with a main body frame of the reinforced fibers to form a spider-screen-frame-imitating toughening structure, so that the strength of the material can be improved, and the toughness of the material is also improved.

The invention described herein relates to a novel non-naturally occurring, elastomeric animal-free recombinant fusion biopolymers produced in plants through transient expression. More specifically, the present invention describes polynucleotides encoding fusion proteins of a non-human scleroprotein with a human collagen, wherein said fusion protein is capable of forming hydroxylated triple helix fibers. In particular fusion proteins of a Spidroin like protein with a human collagen. More in particular, eithera Spidroin-I / Collagen Type-I fusion protein, capable of forming hydroxylated triple helix fibers, or a Fibroin-III / Collagen Type-I fusion protein, capable of forming hydroxylated triple helix fibers. The present invention has improved properties (e.g., thermostability, young's modulus, cell adhesion, degradability, and the like) versus that of native Collagen Type-I. Also described are methods for use thereof, such as the use of electrospun scaffolds which are particularly well suited for biomedical or cosmetic applications as defined in the claims.

The invention relates to the technical field of biology, in particular to recombinant spider silk protein, modified polyurethane and application of the recombinant spider silk protein and the modified polyurethane. The recombinant spider silk protein provided by the invention comprises spider silk protein or spider silk-like protein; the spider silk protein or spider silk-like protein comprises at least one spider silk globulin NT, at least one spider silk protein repeat region and at least one spider silk globulin CT from the N end to the C end. Research finds that when the recombinant spider silk protein is applied to waterborne polyurethane, the mechanical strength of a polyurethane material is remarkably improved, the mechanical property of the polyurethane material is enhanced, and the polyurethane material is environmentally friendly and non-toxic, has the characteristics of no toxicity, high strength, high elasticity and the like, and keeps the original characteristics of low organic matter emission, no pollution, high elasticity, air permeability and the like at the same time. The recombinant spider silk protein can have a CBM structure, so that the mechanical property can be improved. Fluorescent protein and / or chromoprotein can be fused on the recombinant spider silk protein, polyurethane with different appearances can be obtained, and color combination is firm.

The present invention discloses a silk improved by spider silk protein gene and a method for improving silk by spider silk protein gene, wherein the silk improved by spider silk protein gene comprises a silk protein light chain, and the silk protein light chain is connected with a spider silk protein formed by fusion expression with the silk protein light chain; the amino acid sequence of the spider silk protein comprises a repeating region sequence having at least one spider silk protein repeating unit sequence. The present invention realizes the expression of the pear-shaped gland silk of Astilbe punctata in Bombyx mori silk fibroin, realizes the formation of a complete spider silk protein on the silk light chain, and expands the gene editing site of the silk gland of Bombyx mori. This light chain editing silkworm can be crossbred with the heavy chain gene editing silkworm, providing technical support for further breeding and optimization of fiber performance.

The invention discloses a composition containing spider silk hydrolysate. The composition is prepared from the following raw materials in parts by weight: 25-35 parts of spider silk hydrolysate, 15-20 parts of cinnamon leaf residue extracting solution, 2-6 parts of lotus extracting solution, 3-7 parts of stearyl alcohol, 5-9 parts of glycerol, 2-6 parts of palmitoyl tripeptide and 14 parts of stearyl glucoside. The invention further provides application of the composition containing the spider silk hydrolysate in preparation of anti-aging drugs or skin care products. According to the composition of the spider silk hydrolysate, the anti-aging effect is improved, the added stearyl alcohol, glycerol, palmitoyl tripeptide and stearyl glucoside are used as auxiliary materials to be applied to anti-aging drugs or skin care products, the good anti-aging curative effect is achieved, the increase rate of the skin moisture content of the products can be improved due to the addition of the spider silk hydrolysate, and the anti-aging effect of the products is improved. And meanwhile, the anti-aging effect of the product can be stabilized, and the raw materials are matched with one another, so that a remarkable improvement effect is achieved.

A recombinant spider silk protein, consisting of no more than 800 amino acids, comprising a set of domains arranged according to the formula (NT)-REP-CT, wherein: the optional NT-domain, if present, comprises a sequence of 100 to 160 amino-acid residues derived from the N-terminal domain of a spider silk protein; the REP-domain comprises a sequence of 30 to 600 amino acid residues derived from the repetitive segment of a spider silk protein; and the CT-domain comprises a sequence of 70 to 120 amino acid residues derived from the C-terminal domain of a spider silk protein selected from: a sequence of 72 to 110 amino acid residues derived from the C-terminal domain of a spider silk protein, wherein the sequence comprises at least 7 residues independently selected from K, R, E and D; a sequence having at least 85% identity to SEQ ID NO: 15 or any one of SEQ ID NOs: 62-65 or 67-73; and a sequence having at least 70% identity to SEQ ID NOs: 64 or any one of SEQ ID NOs: 62-65 or 67-73, wherein the sequence comprises at least 7 residues independently selected from K, R, E and D.

The invention provides a spider silk protein modified body easy to purify as well as a modification method and a purification method thereof, a plurality of amino acid residues in flexible regions on two sides of a spider silk protein microcrystalline region are replaced with acidic amino acids or alkaline amino acids, so that the proportion of charged amino acids in the flexible regions is greater than or equal to 7%, and the spider silk protein modified body is obtained; the spider silk protein modified body is purified in the form of an inclusion body through a temperature change process of heating and cooling. The modified body can be simply purified with high recovery rate through simple heating and cooling processes, the total recovery rate is up to 98% or above, the purity is 90%-99% or above, and meanwhile, the formed artificial spider thread is endowed with higher tensile strength by strengthening the interaction of beta-sheet on the two sides of a microcrystal region, especially between different chains. A foundation is further laid for large-scale efficient production and low-cost separation and purification of the high-performance spider silk protein, and the application possibility of the spider silk protein in various fields is greatly enhanced.

A plurality of methods and apparatus for managing deficiencies of the meibomian gland lipid production and delivery, including wearable fabrics configured with a matrix of micro- and nano-electromechanical materials, or composite fibres of yarn that are substantially made from carbon nanotubes, graphene, spider silk, natural silk, denim, cotton, metals, or combination thereof. Other methods and apparatus include infrared and ultrasound energy sources within a kit aimed at personalised therapies for improving overall health of the eyelids, while providing for adjunctive relief for associated dry eye symptoms. The management kit may comprise a wearable fabric or spectacle frame configured with infrared emitters and sensors, a handheld imaging device and software containing dosage and administration information coupled with information on improving eyelid hygiene which is reviewed and periodically updated using user-specific information.

The present invention relates to a novel injectable protein delivery system and a method for delivering one or more therapeutic agents to the central nervous system (CNS) to promote axonal regeneration. The system is based on the use of a recombinant spider silk protein, referred to as spiroin-SpyTag, which undergoes a rapid transition from a sol state to a gel state when exposed to ultrasonic treatment and incubated at body temperature. This unique characteristic allows the material to be easily injected into a specific target tissue. The methods disclosed herein can deliver a protein therapeutic agent covalently conjugated to spiroin-SpyTag to a subject affected by a CNS disorder or injury. In addition, methods of making such injectable protein delivery systems are fast, convenient, and economical and effective.

The application provides an aggregated spider silk protein nanoparticle, a drug delivery system and a preparation method and application thereof, and belongs to the technical field of biological medicines.The application provides application of the aggregated spider silk protein in preparation of a drug delivery system.The application researches and finds that the aggregated spider silk protein nanoparticle prepared by using the amino acid sequence such as SEQ ID NO.1, SEQ ID NO.3 or SEQ ID NO.5 has good colloidal stability and good safety, and is a novel drug delivery system.The drug delivery system loaded with a tumor drug has high encapsulation efficiency and drug loading capacity for the tumor drug, the encapsulation efficiency is more than 60%, has a slow-release effect on the tumor drug, and is an excellent drug delivery system.The research finds that compared with the simple tumor drug, the drug delivery system loaded with the tumor drug has a significantly improved killing effect on tumor cells, and can effectively treat tumors.

The invention discloses a natural spider silk-based self-adaptive surgical suture and a preparation method thereof, and belongs to the technical field of biomedical materials. According to the surgical suture, natural spider silk serves as a raw material, protein chains in the spider silk are rearranged in order by regulating and controlling the moisture state, applying axial pre-stretching strain and conducting drying, annealing and shaping under the limited condition, and the suture material with the high strength and the environmental response characteristic is obtained. The surgical suture is based on natural spider silk, has excellent biocompatibility and inherent biodegradability, becomes an ideal candidate material for biomedical application, and is successfully applied to suture of skin, stomach and other parts. According to the invention, the spider silk protein chains are rearranged into the superfine nanofibers, so that the engineered natural spider silk suture with remarkably enhanced mechanical and functional properties can be produced, and the monofilament can realize the breaking strength of 2.0 GPa, the toughness of 480 MJ m < 3 > and the driving stress of 92.6 MPa.

The invention discloses a costume fabric containing spider silk, which is characterized in that modified spider silk is used as first warp and weft spinning yarn, antibacterial cotton thread is used as second warp and weft spinning yarn, and then the first warp and weft spinning yarn and the second warp and weft spinning yarn are interlaced and woven in the warp and weft direction to obtain the costume fabric containing spider silk. In addition, the invention further provides a preparation method of the garment fabric containing the spider silk. The prepared fabric containing spider silk is simple in preparation method and very excellent in breaking strength, tearing strength and puncture resistance, and the fabric has very good antibacterial property on staphylococcus aureus and escherichia coli after being subjected to aging treatment or saline water treatment and is also very excellent in antibacterial stability.

This invention discloses a self-contained conical microchannel and its design method. The microchannel includes a conical channel with a biomimetic fishtail structure and a biomimetic spider silk structure on its surface. The biomimetic fishtail structure is a fishtail-shaped groove formed by bending the base of a triangle with a consistent orientation inward. The biomimetic spider silk structure is a spider silk-shaped groove composed of hemispherical and conical channels. Multiple biomimetic fishtail structures are arranged in an array, and multiple biomimetic spider silk structures are arranged in an array, alternating between the fishtail and spider silk arrays. This invention combines the biomimetic fishtail and spider silk structures on the surface of the conical channel. The fishtail structure enables self-collection of liquid within the conical microchannel, while the spider silk structure enables rapid self-transportation of liquid. This technology can be applied to micro-drug delivery without external force, microfluidic devices, and friction interfaces.

The invention discloses anti-cracking and anti-freezing fiber composite concrete and a preparation method thereof, and relates to the field of concrete. The preparation method comprises the following steps: immersing bionic spider silk into a microbial mineralization solution, adding a photoresponse nano material, and adjusting illumination, so that calcium carbonate crystals are arranged in a honeycomb shape on the surface of the spider silk, bionic spider silk fibers and microbial mineralization layers are alternately deposited, and a'fiber-mineralization layer-fiber 'sandwich structure is constructed. The structure cooperates with a spider silk spiral and honeycomb mineralization layer, the concrete interface strength is greatly improved, shrinkage cracks are effectively inhibited, microorganisms are activated and mineralized in the cracks to repair the cracks, and the concrete is endowed with high crack resistance and self-repairing capacity. Meanwhile, the self-made nano-composite fiber takes polyvinyl alcohol and ethylene glycol copolymer as main materials, polylactic acid micro-cavities containing anhydrous calcium chloride are connected to two ends of the self-made nano-composite fiber, and the freezing resistance of the concrete is improved by the hydrophilicity of the polyvinyl alcohol and the heat release of the anhydrous calcium chloride. The anti-cracking and anti-freezing fiber composite concrete prepared by the invention has the effects of high cracking resistance and high freezing resistance.

The invention discloses a spider silk collecting device and process, and particularly relates to the technical field of spider silk, the spider silk collecting device comprises a telescopic rod composed of an expansion sleeve and two long rods, the head of the telescopic rod is provided with an installation assembly, the front portion of the installation assembly is provided with flannelette fixed through an opening clamp, and the surface of the flannelette is provided with three-dimensional fluff. And collecting the whole cobweb by means of the three-dimensional fluff. According to the spider silk collecting device and technology, in the storage technology, the flannelette attached with spider webs is fixed to the foaming plates through insect needles, spider silk stress and adhesion caused by flannelette bending are effectively avoided, and through the small foaming plates fixed to the four corners of the foaming plates, when a plurality of foaming plates carrying spider silk are placed in a carton, the spider silk collecting effect is good, and the spider silk collecting effect is good. Stable intervals are formed, and extrusion damage during transportation or storage is prevented.

The application provides a kidney-tonifying and anti-fatigue polypeptide composition and a preparation method thereof, belongs to the technical field of medicinal biological peptide complexes, and the composition comprises oyster polypeptide, holothurian polypeptide, spider silk and blue ears grass-pentamerous fruit composite extract, cistanche-dendrobium officinale composite extract, ginseng extract and deer penis extract; the oyster polypeptide is a common enzymolysis product of oyster meat by trypsin and bovine chymotrypsin; the holothurian polypeptide is a common enzymolysis product of holothurian body wall by alkaline protease, collagenase type II and subtilisin; the spider silk and blue ears grass-pentamerous fruit composite extract is a product obtained by the following steps: subjecting spider silk and blue ears grass and pentamerous fruit to enzymolysis, extracting with an ethanol aqueous solution, and purifying with a SA sulfonic acid silica gel strong cation exchange column; the cistanche-dendrobium officinale composite extract is a product obtained by the following steps: subjecting cistanche and dendrobium officinale to enzymolysis, extracting with an ethanol aqueous solution, and purifying with a phenylboronic acid affinity column-polyamide column series connection column; and the composition realizes the synergistic effect of kidney tonification and anti-fatigue.