40 results about "Spider Proteins" patented technology

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.

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.

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 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.

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.

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 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.

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 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.

The invention relates to the technical field of recombinant protein design and synthesis, and particularly discloses a recombinant spider silk protein and a synthesis method thereof. The recombinant spider silk protein provided by the invention is subjected to N-terminal and C-terminal structural domain replacement of different spider species and different spider silk types, intermediate functional domain repetitive sequence large fragment recombination and other optimization designs, and a series of recombinant spider silk proteins are obtained through a cell-free protein synthesis method and prokaryotic fermentation expression; compared with natural spider dragline silk protein or other existing recombinant spider silk protein, the spider silk protein has good composite mechanical performance and high temperature resistance, meanwhile, the molecular weight is small, the total expression quantity and soluble expression are remarkably improved, and a novel raw material and a design synthesis method of the recombinant spider silk protein are provided for development of high-performance biological materials.

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

The invention discloses a method for producing recombinant spider silk protein based on microalgae photosynthetic expression and non-chromatographic purification, the recombinant spider silk protein and application, and belongs to the technical field of recombinant spider silk protein production. The method comprises the following steps: designing a target recombinant spider silk protein structure, taking a spider dragline silk protein repetitive unit as a core, matching with an end-side elastin structural domain, inserting an amber termination codon TAG at a specified glycine site of the repetitive unit, and introducing non-natural amino acid containing a click chemical functional group by virtue of a gene codon extension technology; the method comprises the following steps: artificially synthesizing an optimized gene expression cassette, constructing a chlamydomonas reinhardtii expression system, synchronously constructing an unnatural amino acid insertion system, co-transforming the gene expression cassette and the unnatural amino acid insertion system into chlamydomonas reinhardtii, screening positive clones, carrying out photosynthetic culture induced expression, and carrying out pH-induced reversible phase separation in combination with magnetic nanoparticle click chemical non-chromatographic purification. Finally, the high-purity recombinant spider silk protein is prepared, and a technical basis is provided for industrialization of biological materials.

The invention discloses a spider silk-like hybrid polymer fiber and a preparation method thereof, and belongs to the field of synthetic fibers, and the preparation method comprises the following steps: dissolving, heating and stirring a carboxyl-containing polymer until the carboxyl-containing polymer is completely swelled, and adding an acid solution for adjustment to obtain polymer gel; adding an amino-containing polymer into the polymer gel, heating and uniformly stirring to obtain hybrid gel, and centrifuging the hybrid gel to obtain a hybrid gel spinning precursor stock solution; extruding the hybrid gel spinning precursor stock solution into an anti-solvent coagulating bath, collecting fibers, and obtaining hybrid polymer fibers after moisture absorption is balanced; and carrying out pre-stretching and moisture absorption balance to obtain the spider silk-like hybrid polymer fibers with different uniaxial orientation and crystallinity structures. According to the invention, continuous preparation of the fiber is realized by combining the high-molecular polymer with anti-solvent induced phase separation and pre-stretching strain programming, the method is simple, the raw material source is environment-friendly, and the prepared fiber has comprehensive characteristics similar to spider silk.

The invention provides a preparation method of spider protein synthetic leather, which comprises the following steps: S10, preparing surface layer slurry: selecting spider protein powder with the average particle size D50 of 1.0-10.0 microns, dispersing the spider protein powder in a polar organic solvent for pre-infiltration treatment to obtain suspension liquid, and drying the suspension liquid to obtain the surface layer slurry; then mixing the suspension liquid with a polyurethane surface layer resin solution, and uniformly dispersing to prepare surface layer slurry containing spider protein powder; s20, preparing inner-layer slurry: preparing polyurethane inner-layer slurry which does not contain arachnoid protein powder; s30, dry molding: coating release paper with the surface layer slurry, drying the surface layer slurry to form a surface layer, and controlling the ratio of the dry film thickness of the surface layer to the average particle size D50 of the spider protein powder to be (5: 1)-(10: 1); and then coating the surface layer with the inner layer slurry, drying to form an inner layer, attaching base cloth to the inner layer, and curing and stripping to obtain the spider protein synthetic leather. The synthetic leather prepared by the preparation method has good mechanical properties, good breathable effect and excellent low-temperature folding resistance.

The present application relates to the technical field of water treatment, and particularly relates to a water purifying agent based on a biomimetic adsorption principle and a preparation process thereof, wherein the water purifying agent is obtained by a composite framework of an inner layer, an intermediate layer and an outer layer; the inner layer is a polydopamine nanofiber network; the intermediate layer is a spider silk protein-paraffin composite layer; and the outer layer is a spider silk protein-polydopamine macroporous framework structure; through the multi-layer composite structure, the spider silk protein-polydopamine macroporous framework is the outer layer, the spider silk protein-paraffin composite layer is the intermediate layer, and the polydopamine nanofiber network is the inner layer, so that gradient capture of pollutants from micron to nanometer is realized, and through biomimetic tough framework design, dynamic pore size regulation mechanism, hierarchical adsorption synergistic effect and biomimetic adhesion enhancement technology, the limitation of traditional adsorbents is broken through.

The invention discloses a recombinant expression and in-vitro splicing integrated preparation method and application of spider silk protein, and belongs to the technical field of biology. Coding genes of NT, Rep or CT regions of spider silk protein from at least two different spider species are fused, a fusion gene sequence is optimized by pichia pastoris codon and then introduced into an Intein tag, and the spider silk protein is obtained. Dividing the full-length protein into an N-terminal fragment and a C-terminal fragment; respectively cloning the N-terminal fragment and the Intein-N structural domain as well as the C-terminal fragment and the Intein-C structural domain into a pichia pastoris secreting type expression vector pPICZalpha; respectively electrically transforming the two obtained vectors into pichia pastoris SMD1168 for induced expression, and collecting supernatant; sequentially carrying out Ni-NTA affinity chromatography and ion exchange chromatography on the supernatant to obtain a purified N-terminal protein fragment and a purified C-terminal protein fragment; mixing the spider silk protein and the spider silk protein, forming full-length spider silk protein through Intein-mediated trans-splicing, and purifying through size exclusion chromatography. The method provided by the invention can realize high-efficiency and high-quality production of spider silk protein.

The invention relates to the technical field of leather materials, and particularly discloses wear-resistant leather with an enhanced protein bionic structure and a preparation method of the wear-resistant leather. The wear-resistant leather sequentially comprises a multi-stage bionic wear-resistant surface layer, a protein bionic core layer and base cloth from top to bottom, pangolin scale macroscopic ribs and sharkskin microcosmic grooves are re-carved on the multi-stage bionic wear-resistant surface layer, and a nano hard-phase filler is compounded; the protein bionic core layer adopts a spider silk multistage fiber structure and a shell hard phase embedding mechanism, and a dynamic cross-linked network is introduced; and the transition layer and the dynamic cross-linking bonding layer realize interlayer stable combination and micro-damage self-repairing. The preparation method comprises the steps of raw material pretreatment, raw material preparation of each layer, step-by-step forming, composite curing and the like. Through multistage collaborative bionic and dynamic crosslinking design, the problems that traditional protein-based leather is poor in wear resistance and insufficient in long-term effect are solved, meanwhile, the skin-friendly and environment-friendly advantages are reserved, the wear resistance, the tearing strength and the interlayer binding force are remarkably improved, and the protein-based leather is suitable for high-end shoes, automobile interiors and other scenes.

The invention belongs to the technical field of material science, and particularly provides a preparation method of fiber containing spider silk protein, which comprises the following steps: 1) stirring and mixing 20-50 parts by weight of spider silk protein hydrolysate, 5-10 parts by weight of modified protamine, 20-50 parts by weight of polylactic acid and 20-50 parts by weight of ethanol to form a spinning solution; and (2) spraying the spinning solution obtained in the step (1) into bath lotion by adopting an electrostatic spinning technology, and then performing silk guiding, heating and winding forming to obtain the fiber containing the spider silk protein. Meanwhile, the invention also provides the fiber containing the spider silk protein prepared by the method, and application of the fiber in biomedical materials. The invention mainly solves the problems of single function and insufficient comprehensive performance of the existing fiber containing spider silk protein.

The invention relates to a spider silk protein membrane material capable of generating electricity by friction and a preparation method and application 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 basic amino acids, and spider silk protein modifiers with different flexible region acidic / basic amino acid molar ratios are obtained; the spider silk protein modified body is independently formed or mixed with a conductive material to form a membrane, and the membrane material is formed. By utilizing the method disclosed by the invention, materials needing different piezoelectric properties can be developed, and a foundation is further laid for the industrial production of the recombinant spider silk protein and the wide application of the recombinant spider silk protein in the field of functional materials.

The present disclosure relates to methods for isolating and purifying synthetic block copolymer proteins using solvent and high shear microfluidization techniques.

The invention relates to a recombinant spider silk protein fusion protein as well as a preparation method and application thereof. The fusion protein comprises a VNP6 tag and a recombinant spider silk protein, and the VNP6 tag is located at the N end of the fusion protein. A specific fusion protein structure is designed, and the intracellular space is regulated and controlled by using a VNP6 tag, so that the molecular congestion is effectively relieved, the protein expression yield is improved, and a brand new technical path is provided for improving the expression level of target protein, correctly folding and assembling supramolecular structures (such as artificial spider silk fibers, hydrogel and microneedles).

The invention provides an enterokinase light chain recombinant protein and application and a method for efficient soluble expression of a recombinant enterokinase light chain, the enterokinase light chain recombinant protein is composed of more than two fusion proteins AgSg or mutants thereof and an enterokinase light chain or mutants thereof, the amino acid sequence of the fusion proteins AgSg is shown as SEQ ID NO.3, and the amino acid sequence of the fusion proteins AgSg is shown as SEQ ID NO.3. The amino acid sequence of the enterokinase light chain is as shown in SEQ ID NO. 1; according to the enterokinase light chain recombinant protein, a specific amount of fusion protein AgSg derived from the N end of spider silk sericin is fused with an enterokinase light chain or a mutant thereof in a series connection manner, so that the expression quantity of the enterokinase light chain or / and the mutant can be remarkably improved, and the enterokinase light chain or / and the mutant can be expressed in a soluble form; the defects that a traditional escherichia coli system is low in expression quantity and expression and purification are difficult in an inclusion body form are overcome, and the method can be applied to production of high-efficiency soluble enterokinase light chains.

1. Name of the designed product: transmitter (small spider silk). 2. Use of the designed product: for toys. 3. Design points of the designed product: in shape. 4. Picture or photo best indicating the design points: perspective view.

The present invention relates to recombinant spidroin bioconjugates (chemically modified engineered spider silk proteins). More particularly, the invention relates to recombinant spidroin bioconjugates with poly-ethyleneglycol polymers and their use in regenerative medicine.

The application provides a cysteine-strengthened recombinant spider silk and a preparation method and application thereof. An amino acid sequence of the recombinant spider silk protein has the sequence shown in SEQ ID NO: 6. A polynucleotide encoding the amino acid sequence of the protein has the following structure: a 5' end of a cysteine-modified MaSp2 gene of the major ampullate gland of the spider Argiope bruennichi is connected with an N-domain, and a 3' end is connected with a C-domain; the gene is inserted into a pET-28a plasmid vector to obtain a recombinant plasmid, and the recombinant plasmid is introduced into E. coli to express the recombinant spider silk protein. The recombinant spider silk prepared by wet spinning using the recombinant spider silk protein has a dense beta-sheet crystal structure, excellent mechanical properties and wet strength, and opens up a prospect for industrial application of the recombinant spider silk.