826 results about "Signal peptide" patented technology

The invention belongs to the field of biological medicines, and particularly relates to a varicella-zoster virus gE fusion protein as well as a preparation method and application thereof. The varicella-zoster virus gE fusion protein comprises an amino acid sequence as shown in SEQ ID NO: 1 and at least one of the following fragments: a signal peptide, a Foldon sequence, a Ferritin sequence and a His tag sequence, the signal peptide can improve the protein expression quantity, the Foldon sequence can improve the stability of the protein, the particle size of an antigen can be increased, the immunogenicity of the antigen can be improved, and the fusion protein can be used for preparing a medicine for treating the varicella-zoster virus gE fusion protein. The Ferritin sequence can enhance immune response, a vaccine prepared from the gE fusion protein has excellent immunogenicity, and compared with a commercially available vaccine Shingrix, higher humoral immunity and cellular immunity levels can be induced.

The invention discloses a CA9-targeted chimeric antigen receptor and application thereof, and belongs to the technical field of biological medicines. The CA9 targeting chimeric antigen receptor provided by the invention comprises a CD8 alpha leading signal peptide, an anti-CA9 single-chain variable fragment, a CD8 alpha hinge, a CD8 transmembrane structural domain and a CD3 zeta intracellular costimulatory signal transduction structural domain. According to the invention, an anti-CA9 single-chain variable fragment is used as an antigen binding structural domain to construct a chimeric antigen receptor (CAR) molecule, the macrophage expressing the anti-CA9CAR can realize specific targeting and phagocytosis killing of CA9 positive kidney cancer cells, and the constructed lipid nanoparticle delivery system can effectively transfer a gene expression vector, so that the gene expression vector can realize effective target site delivery, and the CA9 positive kidney cancer cells can be effectively targeted and killed. The constructed hydrogel-liposome combined drug delivery system has the performance of relieving toxic and side effects, controllable release and hemostasis.

The invention belongs to the field of skin care products, and particularly relates to an anti-wrinkle and anti-aging composition as well as a preparation method and application thereof, and the anti-wrinkle and anti-aging composition comprises neurotransmitter inhibition peptide, signal peptide, madecassoside, inositol and glycosylglycerol. The neurotransmitter inhibition type peptide is at least three of acetyl hexapeptide-8, arginine / lysine polypeptide, dipeptide diaminobutyrylbenzylamide diacetate and acetyl octapeptide-3, and the signal type peptide is at least two of palmitoyl pentapeptide-4, decapeptide-4 and acetyl tetrapeptide-9. Compared with the prior art, all the components of the anti-wrinkle and anti-aging composition act on different anti-aging pathways and can be matched with one another from multiple aspects of oxidation resistance, inflammation resistance, moisturizing, cell viability enhancement, wrinkle reduction and the like, a more comprehensive and better anti-aging effect is provided, skin aging is comprehensively resisted, the skin is in a more young state, and the anti-wrinkle and anti-aging composition has the advantages that the anti-wrinkle and anti-aging effects are achieved. And the anti-wrinkle and anti-aging composition is small in irritation and good in stability.

The invention relates to the field of cell biology, and particularly discloses an exosome as well as a preparation method and application thereof. The method for preparing the exosome comprises the following steps: (1) introducing nucleotide sequences for coding a signal peptide TPA and an epidermal growth factor into an AAVS1 region of a host cell, such as a sequence shown in SEQ ID No.9 and a sequence shown in SEQ ID No.10, so as to obtain a modified host cell; and (2) collecting the exosome secreted by the modified host cell. The expression level of EGF mRNA in the exosome can be remarkably improved, so that the obtained exosome has a better effect in skin wound treatment.

The invention belongs to the field of biological medicine, and particularly relates to a respiratory syncytial virus F protein mutant and application thereof. The respiratory syncytial virus F protein mutant is characterized in that at least one pair of amino acid residues in an F1 subunit and / or an F2 subunit of F protein polypeptide are substituted by cysteine, and / or an F protein signal peptide part is substituted by a human immunoglobulin light chain signal peptide, and / or alanine A at the 107 site of a multi-base sequence is mutated into arginine R, lysine K at the 134 site is mutated into arginine R, and the amino acid residues in the F1 subunit and / or the F2 subunit of the F protein polypeptide are mutated into arginine. According to the invention, more neutralizing antibody epitopes can be exposed, the stable pre-fusion conformation is maintained, effective neutralizing antibody reaction and antibody binding reaction to the RSV subtypes A and B can be ensured, and the vaccine is suitable for various vaccine forms taking the RSV F protein as the antigen.

The invention relates to the technical field of biology, in particular to a pseudorabies virus gD recombinant protein, a subunit vaccine and a preparation method and application thereof.The gD recombinant protein sequentially comprises a signal peptide, a connecting peptide, the 21st-350th site of the gD protein, an enzyme cutting site and a His tag from the N end to the C end, the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.4, and the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.6. The pseudorabies virus gD recombinant protein has high immunogenicity, the prepared subunit vaccine can generate a specific antibody with neutralizing activity, mice do not die after challenge, the protection rate is 100%, the protection rate after challenge of pigs is 100%, pathological sections only show slight pathological changes, and the pseudorabies virus gD recombinant protein has wide application prospects.

The invention discloses a group of mitochondrial signal peptides and application thereof, and a group of signal peptides with a mitochondrial targeting function is obtained for the first time by screening endogenous mitochondrial key functional proteins of rhodotorula. The invention further realizes expression of a target gene through mitochondrial signal peptide mediation, realizes expression of a target product synthetic pathway gene in rhodotorula mitochondria, and successfully constructs synthetic pathways of terpene such as limonene, linalool, patchouli alcohol and kaurene, branched chain alcohol such as isobutanol and organic acid such as 3-hydracrylic acid. Further, carbon metabolic flow of pyruvic acid flowing to the target product is dredged, and synthesis of the mitochondrial region indoor target product is finally realized. The characterization of a group of rhodotorula mitochondrial signal peptides disclosed by the invention can realize the synthesis of mitochondrial regional chamber target products, and lays a foundation for the research on the realization of metabolic engineering transformation in subcellular organelles.

The invention belongs to the technical field of biological medicine, and particularly relates to a preparation method of viroid particles displaying scFv antibodies, a product of the viroid particles and application of the viroid particles. The preparation method provided by the invention comprises the following steps: constructing a specific targeting recombinant plasmid, wherein the specific targeting recombinant plasmid is obtained by connecting a vector for expressing virus capsid protein and a coding gene for expressing an scFv antibody display structure through homologous recombination. The vector for expressing the virus capsid protein comprises a coding gene of VSV-G signal peptide, a transmembrane region and an intracellular region, and is as shown in SEQ ID NO. 2. And the coding gene for expressing the scFv antibody display structure is shown as SEQ ID NO. 3-5. According to the preparation method disclosed by the invention, the virus-like particles of which the capsid is a modular component of a specific targeting immune cell are constructed based on a lentivirus system, and 50-90% of infection efficiency can be realized in the immune cell.

In one embodiment, the present disclosure provides an engineered cell having a first chimeric antigen receptor polypeptide including a first antigen recognition domain, a first signal peptide, a first hinge region, a first transmembrane domain, a first co-stimulatory domain, and a first signaling domain; and a second chimeric antigen receptor polypeptide including a second antigen recognition domain, a second signal peptide, a second hinge region, a second transmembrane domain, a second co-stimulatory domain, and a second signaling domain; wherein the first antigen recognition domain is different than the second antigen recognition domain.

The present invention discloses a method for producing protein HFBI. The method comprises the following steps: infecting insect cells with a recombinant baculovirus containing an insect cell expression vector expressing a fusion protein, and collecting the supernatant; the fusion protein is composed of a GP64 signal peptide and a protein HFBI; the amino acid sequence of the GP64 signal peptide is shown in SEQ ID NO:2 from the 1st to the 20th position from the N-terminus; then the supernatant is precipitated with ammonium sulfate to obtain a large amount of protein HFBI, and finally the protein is further purified by heating using the thermal stability. The purity of the protein HFBI obtained by the method provided by the present invention is more than 95%, the concentration is 6 mg / ml, and it has biological activity. It can be seen that the present invention can significantly increase the yield of protein HFBI, and the production process is simple, which can be fully applied to actual production. The present invention has important application value.

Disclosed are a recombinant fusion antibody, and an antibody-drug conjugate and use thereof. The recombinant fusion antibody includes an antibody, at least one lysosome-targeting peptide, and at least one linker peptide, wherein the antibody is fused with the lysosome-targeting peptide to form the recombinant fusion antibody via the linker peptide; wherein the lysosome-targeting peptide is at least one selected from the group consisting of a lysosome-sorting signal peptide from a Golgi-localized, γ-adaptin ear-containing, ARF-binding protein (GGA), a lysosome-sorting signal peptide from a mannose 6-phosphate receptor (MPR), a lysosome-sorting signal peptide from a lysosome-associated membrane protein (LAMP), and a variant; wherein the variant is a peptide with one or more amino acids substituted, deleted, and / or added when aligned with the lysosome-sorting signal peptide from a GGA, a MPR or a LAMP. Compared with the original antibody, the internalization and lysosomal trafficking ability of the recombinant fusion antibody are significantly improved.

The present invention provides a plant expression method of type III human collagen, an intron-containing coding gene and its application. The method comprises: step A: adding intron fragments to 16 gene fragments of collagen to construct the collagen COL3A1 gene; step B: connecting and synthesizing, and at the same time, achieving type III human collagen targeting to the apoplast by adding a signal peptide to achieve extracellular expression of type III human collagen; step C: transgenic expression of collagen; step D: culturing and purifying the best expressing plant seeds in step C. The present invention expresses type III human collagen in the apoplast, simplifies the purification steps, reduces costs, and the optimization of adding introns significantly enhances the stability of collagen expression.

The invention discloses a method for promoting lactobacillus activity enhancement, which comprises the following steps: S1, amplifying a target gene corresponding to an LPxTG structural protein, and amplifying a target gene corresponding to a signal peptide; s2, obtaining a target gene connected with the LPxTG structure and the signal peptide; s3, connecting the target gene obtained in the step S2 with a plasmid gene sequence to construct a recombinant expression vector; s4, introducing the recombinant expression vector obtained in the step S3 into escherichia coli BL-21 so as to increase the number of the recombinant expression vector; s5, introducing the recombinant expression vector obtained in the step S4 into lactobacillus plantarum C8 to obtain a recombinant strain; and step S6, co-culturing lactobacillus plantarum C8 and the recombinant strain obtained in the step S5. The lactobacillus plantarum C8 has the advantages that the whole lactobacillus plantarum C8 and the recombinant strain have better survival condition and adhesiveness in gastric juice and intestinal juice.

The invention discloses a recombinant protein for detecting a para-tumor Yo antibody through a CBA method and application, and belongs to the technical field of biomedical engineering.The amino acid sequence of the recombinant protein sequentially comprises a secretory signal peptide, a CDR2 protein partial sequence, a CDR2L protein partial sequence, a transmembrane region and a fluorescent label, and the secretory signal peptide is human albumin signal peptide ALB; the transmembrane region is a CD8a hinge; and the fluorescent label is mCherry. A novel recombinant protein which can be stably over-expressed on a cell membrane of an eukaryotic cell is constructed by intercepting specific partial sequences of CDR2 protein and CDR2L protein and redesigning and fusing a fluorescent label by using a secretory signal peptide, a transmembrane sequence and a connecting peptide, and the recombinant protein retains respective core antigen regions of the CDR2 protein and the CDR2L protein, so that the specific partial sequences of the CDR2 protein and the CDR2L protein can be stably over-expressed on the cell membrane of the eukaryotic cell. The kit can effectively overcome the defects in the aspects of sensitivity and specificity, and when a CBA method is adopted for detection, the detection rate of the para-tumor Yo antibody can be remarkably increased, and the false positive rate is reduced, so that the requirements of clinical detection are better met.

The invention relates to the technical field of active peptides, in particular to an active peptide composition and application thereof in preparation of anti-aging and wrinkle-removing products. The composition provided by the invention comprises the neurotransmitter regulating peptide and the signal peptide, and the components are properly selected, so that the composition can play a better role in anti-aging and anti-wrinkle aspects. In addition, the oil-soluble wrapped anti-aging and anti-wrinkle composition prepared by the invention aims to be quickly absorbed through skin, and the utilization rate of the active polypeptide is effectively improved.

The invention relates to the technical field of gene engineering, in particular to Escherichia coli for efficient soluble expression of recombinant human collagen III and construction and application thereof. The invention constructs Escherichia coli for efficient soluble expression of recombinant human III type collagen, and the Escherichia coli can co-express the recombinant human III type collagen and molecular chaperone which are regulated and expressed by signal peptide. Experiments prove that the engineering bacterium provided by the invention has the capability of efficient soluble expression of the recombinant human III-type collagen, and the recombinant human III-type collagen provided by the invention has good stability and anti-inflammatory effect.

The invention discloses bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis. The invention provides bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis in fat cell extraction, and after optimization of an expression element promoter and a signal peptide and large-scale fermentation of a fermentation tank, the highest enzyme activity can reach 669U. ML <-1 >. After purification, adipose-derived cells are successfully extracted and separated from mouse adipose cells, and after passage, the adipose-derived cells present the form of adipose cells. The method shows that the method has a good application prospect in the aspect of fat cell extraction.

The invention discloses a high-yield delta-tocotrienol genetically engineered bacterium and application thereof, and belongs to the technical field of gene recombination. The invention discloses a genetically engineered bacterium for high yield of delta-tocotrienols, which is characterized in that yarrowia lipolytica ATCC MYA-2613 is used as an original strain, and a ku70 gene is knocked out; the gene is used for expressing and coding 4-hydroxyphenylpyruvate dioxygenase, the gene is used for coding uromelanic acid phytate transferase and the gene is used for coding tocopherol cyclase; intensifying a shikimic acid pathway; a mevalonic acid pathway is strengthened; an isopentenol path is introduced; carrying out protein fusion on the HPD and the HPT; truncating a signal peptide of a gene encoding a tocopherol cyclase and back-filling a leucine defect. The genetically engineered bacterium constructed by the invention has industrial stability, can be amplified by more than 10 times in a 2L fermentation tank level compared with a shake flask level, can produce 616.45 mg / L of delta-tocotrienol after being cultured for 240 hours, and has industrial application prospects.

The invention discloses an E-CAR-NK cell for treating systemic lupus erythematosus and application of the E-CAR-NK cell, and belongs to the technical field of genetic engineering.A CD19 and BCMA double-target E-CAR is adopted for modifying an NK cell, and the E-CAR-NK cell is obtained; the CD19 and BCMA double-target E-CAR is composed of the following modules: a CD8 signal peptide, a CD19 scFv, a Linker, a BCMA scFv, a CD8 Hinge region, an NKG2D transmembrane region, a CD3 epsilon signal transduction region, a DAP12 costimulatory region, a 2B4 costimulatory region, and a CD3 zeta signal transduction region. Compared with a common CAR-NK cell, the E-CAR-NK cell disclosed by the invention has higher antigen sensitivity and better continuous killing ability, so that the treatment effect of SLE is improved, and the E-CAR-NK cell has lower side effects and higher safety.

The present invention relates to a bispecific chimeric antigen receptor targeting CD117 and CD7 and its application, wherein the bispecific chimeric antigen receptor comprises a signal peptide, an antigen binding domain, a hinge region, a transmembrane domain and a signal transduction domain, wherein the antigen binding domain comprises a CD117 binding domain and a CD7 binding domain. The bispecific chimeric antigen receptor provided by the present invention combines targeted antibodies for CD117 and CD7 together, recognizes and binds to two different antigens at the same time, enhances the targeted elimination ability of malignant cells, and by acting on hematopoietic stem cells and immune cells at the same time, can more effectively eliminate tumor cells in the receptor, while retaining or reconstructing normal hematopoietic and immune functions.

The invention discloses a genetically engineered bacterium for producing tanshinol as well as a construction method and application of the genetically engineered bacterium, and the construction method comprises the following steps: (1) taking escherichia coli W3110 as an original strain, and reconstructing a tanshinol synthesis route in cytoplasm; (2) positioning flavin reductase from klebsiella pneumoniae and a 4-hydroxyphenylacetic acid-3 hydroxylase mutant in a periplasmic space by using periplasmic space positioning signal peptide sufI. The genetically engineered bacterium for producing the tanshinol disclosed by the invention has the advantages that the yield of the tanshinol reaches 2.8 g / L after being subjected to shake flask fermentation for 24 hours, and the yield of the tanshinol reaches 26.6 g / L after being fermented in a 5L fermentation tank for 48 hours.

The invention provides a saccharomyces cerevisiae engineering strain HYB05 capable of releasing astaxanthin in a controllable self-splitting manner. The saccharomyces cerevisiae engineering strain HYB05 is constructed by taking a basic strain for producing astaxanthin as a starting strain, introducing a coding sequence comprising cell wall lyase Lyc, a coding sequence of a signal peptide mutant F8A and a recombinant nucleic acid construct for expressing a regulatory element, and knocking out an SSD1 gene to relieve a translation inhibition effect. The cell wall lyase Lyc is found and disclosed by the inventor for the first time, and the amino acid sequence is shown as SEQ ID NO: 6; the amino acid sequence of the signal peptide mutant F8A is as shown in SEQ ID NO: 8. According to the engineering strain HYB05, by introducing the recombinant nucleic acid construct, the activation of the catalytic activity of cell wall lyase under specific conditions is realized, so that the cell wall of the saccharomyces cerevisiae is broken, and the astaxanthin synthesized by the saccharomyces cerevisiae is released. Compared with the prior art, the astaxanthin is produced through fermentation of the strain, the extraction steps are simplified, extraction of the astaxanthin in the yeast is achieved to the maximum extent, the yield of the astaxanthin is remarkably increased, and the technical effect is remarkable.

The present invention relates to a viral gene therapy vector for removing hepatitis B viruses, a preparation method therefor, and the use thereof, aiming to solve the problem in which existing viral gene therapy vectors can only inhibit the replication of hepatitis B viruses, but cannot remove virus cccDNA. The vector comprises a promoter, an intron, an enhancer, a nuclear localization signal (NLS) functional domain encoding gene of PGLYRP2 protein, an HBV DNA binding domain encoding gene of the PGLYRP2 protein, a secretory signal peptide functional domain encoding gene of the PGLYRP2 protein, an HBV nucleocapsid-binding functional domain encoding gene of the PGLYRP2 protein, and an IRES fragment. The viral gene therapy vector can remarkably promote the removal of HBVs in the liver of mice and is used for treatment of hepatitis B virus infection.

The invention relates to a design method of a CAR-T cell model targeting human TROP2 and application of the CAR-T cell model in triple negative breast cancer. The method comprises the following steps: obtaining protein sequences of a light chain and a heavy chain by utilizing a sequence of a targeting drug of the human TROP2; comparing the protein sequences of the light chain and the heavy chain with the sequence of the drug protein, and removing the same sequence part to obtain a variable protein sequence; transforming the variable protein sequence into a base sequence to respectively obtain base sequences of a light chain variable region and a heavy chain variable region, and adding a signal peptide and a connecting region to synthesize two base sequences to obtain a human TROP2 antibody sequence fragment; the preparation method comprises the following steps: synthesizing plasmids carrying a human TROP2 antibody sequence fragment by taking an SFG retroviral vector as a skeleton; and enabling the human T cells to express a target gene through transfection. In the embodiment, the nucleic acid sequence of the human TROP2 antibody as shown in SEQ ID NO.1 and SFG are used as carriers, a retrovirus transfection system is used for constructing a TROP2. CAR-T cell model, the functionality of the TROP2. CAR-T cell on the triple negative breast cancer is defined, and a new method is provided for clinical treatment of the triple negative breast cancer.

The present invention discloses an improved polypeptide tumor vaccine composition and application, specifically relating to the field of polypeptide tumor vaccines, the vaccine comprises a nucleic acid sequence encoding a plurality of mutant tumor-associated antigens, the nucleic acid sequence including but not limited to: a nucleic acid sequence encoding a mutant WT1 antigen, selected from the amino acid sequence of SEQ ID NO: 1 or a variant thereof with at least 90% identity; a nucleic acid sequence encoding a mutant MUC1 antigen, selected from the amino acid sequence of SEQ ID NO: 2 or a variant thereof with at least 90% identity; a nucleic acid sequence encoding a mutant PD-1 antigen, selected from the amino acid sequence of SEQ ID NO: 3 or a variant thereof with at least 90% identity; and the vaccine composition further comprises one or more modified elements that enhance antigen expression or immune response, including a signal peptide, an adjuvant coding sequence or an immunostimulatory sequence. The vaccine composition of the present invention is intended to enhance the immune response to a plurality of tumor antigens and improve the effectiveness of tumor treatment.

The invention provides a classical swine fever virus subunit vaccine and a preparation method thereof, the classical swine fever virus subunit vaccine is based on an E2 protein gene and a GP67 signal peptide gene of a CSFV 1.1 strain, a recombinant expression vector is constructed through gene cloning, a classical swine fever virus E2 recombinant protein is obtained through expression of an insect baculovirus system, and the classical swine fever virus subunit vaccine is obtained. And the recombinant protein is combined with a colloidal manganese adjuvant to prepare the swine fever virus subunit vaccine. The hog cholera virus E2 recombinant protein is close to natural E2 protein in structural function, high in reduction degree, high in expression quantity and suitable for vaccine production and application; the vaccine can generate specific humoral immunity and relatively strong cellular immunity, and is beneficial to removal of CSFV in an animal body; meanwhile, the vaccine can be distinguished from attenuated vaccine strain immunization or wild strain infection at the nucleic acid level, can be used for preventing and purifying classical swine fever in an intensive pig farm, and has a considerable application prospect.

The invention relates to a high-yield trans-aconitic acid engineering bacterium as well as a construction method and application thereof, and belongs to the technical field of gene engineering. In the scheme of the invention, the inventor introduces mitochondrial localization signal peptide to be combined with the aconitic acid isomerase to form a gene element, and an expression cassette containing the gene element is introduced into the strain by adopting a genetic engineering technical means, so that the aconitic acid isomerase expressed in the constructed recombinant strain can be localized into mitochondria, and the expression efficiency of the recombinant strain is improved. Metabolic pathways related to trans-aconitic acid synthesis are located in the same zone, so that the capacity of fermenting and producing trans-aconitic acid by the recombinant strain is improved. The technical scheme disclosed by the invention not only has a good application prospect and production economic value, but also provides a referential new thought for transformation of strains related to other fermentation processes.

The present invention belongs to the technical field of gene therapy. Provided are a chimeric antigen receptor nanocomposite and a preparation method therefor and the use thereof in the preparation of an anti-aging drug. The provided chimeric antigen receptor nanocomposite comprises a chimeric antigen receptor DNA plasmid specifically recognizing senescent cells, a polymer coupled with a signal peptide, a free polymer and an antibody polymer targeting T cells, wherein the chimeric antigen receptor DNA plasmid, the polymer coupled with a signal peptide, the free polymer and the antibody polymer targeting T cells are sequentially mixed and incubated to form the chimeric antigen receptor nanocomposite. The chimeric antigen receptor nanocomposite can rapidly generate senescent cell-marked chimeric antigen T cells in vivo. The engineered T cells have the ability to specifically eliminate senescent cells in vivo, and can rapidly and durably eliminate senescent cells in vivo, which is irreplaceable and has wide application prospects.

The invention relates to the technical field of antibacterial peptides, and discloses a preparation method and application of a multi-segment fused antibacterial peptide fusion protein. The antibacterial peptide fusion protein is obtained by introducing OST signal peptide and collagen and optimizing the connection sequence of the OST signal peptide, the collagen, bovine lactoferrin peptide LFcinB, bovine lactoferrin N-leaf small peptide LFcin and bovine lactoferrin N-leaf derived peptide Lf. The antibacterial peptide fusion protein can simultaneously contain three segments of bovine lactoferrin active peptides, namely LFcinB, LFcin and Lf, and the yield of the fusion peptide is maximized. Experiments prove that the fermentation yield of the fusion protein comprising the OST signal peptide, the collagen, the bovine lactoferrin peptide LFcinB, the bovine lactoferrin N-leaf small peptide LFcin and the bovine lactoferrin N-leaf derived peptide Lf which are sequentially connected from the N terminal to the C terminal can reach 600mg / L.

The invention relates to the technical field of bioengineering, in particular to recombinant saccharomyces cerevisiae for producing a human epidermal growth factor based on a biological membrane as well as a construction method and application of the recombinant saccharomyces cerevisiae. According to the invention, saccharomyces cerevisiae BY4742 and 530-1C are used as chassis cells for expressing the human epidermal growth factor (hEGF), and the yield of the human epidermal growth factor (hEGF) is increased to 15 mg / L from 80 [mu] g / L by means of gene modification, promoter optimization, signal peptide optimization and the like; by optimizing related modules in an engineering protein secretion pathway, single or combined deletion of degrading enzyme genes (PEP4, PRB1 and YAP3) and overexpression of key node genes (KAR2, PDI1, HAC1, ERV29, ERO1, SRP54, SEC24 and DER1) in the protein secretion pathway, the hEGF yield is increased to the level of 141 mg / L; a biological membrane strain with the performance of flocculation and adhesion to an abiotic surface is constructed by selecting an endogenous flocculation gene FLO11 of overexpression yeast and an exogenous gene ALS3 of tritirachium album, and continuous production of hEGF is realized based on biological membrane immobilized continuous fermentation, and the yield is stabilized at 300 mg / L or above.