109 results about "Secretion expression" patented technology

The invention discloses recombinant human VII type collagen as well as a preparation method and application thereof, and belongs to the technical field of biochemical engineering. The recombinant human VII type collagen comprises a plurality of amino acid residue repetitive units shown as SEQ ID NO.1. Compared with natural VII type collagen, the recombinant human VII type collagen obtained by repeating the repetitive units for multiple times has the advantages that the molecular weight is smaller, the sequence is more easily and efficiently secreted and expressed in a pichia pastoris system after optimization, and the recombinant human VII type collagen can be used for preparing the recombinant human VII type collagen. The method has the characteristics of high yield, low cost, simplicity in operation and easiness in large-scale production. Meanwhile, the anti-oxidation activity is excellent, the skin moisturizing and repairing capacity and the wound healing capacity are promoted, due to the fact that the collagen is derived from human collagen, the risks of allergy and rejection reaction can be remarkably reduced, and the safety is extremely high. Therefore, the recombinant collagen can be used as a core active component to be widely applied to the fields of cosmetics, medicines and the like, and has a good application prospect.

The invention discloses recombinant poria cocos chitin endonuclease as well as a gene, a preparation method and application thereof. The nucleotide sequence of the gene is shown as SEQ ID NO.1 in a sequence table. According to the nucleotide sequence as shown in SEQ ID NO.1, high-level recombinant secretory expression of target protein in pichia pastoris host bacteria can be realized by taking pichia pastoris inducible expression plasmid pPICZ alpha A as a carrier. Under a shake flask fermentation condition, the average secretory expression quantity can reach 300.3 mg / L. The recombinant enzyme obtained by nickel ion affinity chromatography purification not only can effectively hydrolyze colloidal chitin to generate chitosan oligosaccharide (COS), but also can directly degrade chitin-containing raw materials such as shrimps, crabs and the like to generate COS. In addition, the enzyme also shows the activity of hydrolyzing polysaccharides such as cellulose, pectin, agar and the like, and has important potential values in the fields of functional chitosan oligosaccharide preparation, biological energy production, medicine development, feed, environmental protection, agricultural application and the like.

The present disclosure relates to chimeric antigen receptor cells targeting ROBO1, in particular, enhanced CAR-T cells and CAR-NK cells targeting ROBO1, and preparation and application thereof. The cells can stably expressing CAR elements, while secreting extracellular domain molecules expressing PD-1 protein or mutants thereof, and thus may block PD-11PD-L1 molecular interaction. It has been found through animal experiments that the cells have very good anti-tumor effects, and the above-mentioned cells can significantly reduce tumor recurrence and improve the survival rate compared with the conventional ROBO1-targeted CAR modified cells.

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 method for conformational signal peptide-mediated secretion and expression of heterologous proteins in Escherichia coli, and specifically comprises the following steps: simultaneously fusing at least two heterologous proteins to a conformational signal peptide (such as sfGFP) and constructing a heterologous recombinant protein expression vector, wherein the conformational signal peptide has a beta-barrel structure, and the fusion sites are at least two of the amino terminal end, the carboxyl terminal end and a plurality of loop rings of the conformational signal peptide; and then transforming the heterologous recombinant protein expression vector into Escherichia coli and culturing the obtained genetic engineering strain for expression. According to the method, the extracellular secretion and expression of the heterologous proteins in the Escherichia coli can be realized without the mediation of a classical signal peptide, the sfGFP as a fusion tag has no influence on the function of the target protein, and the corresponding target protein can be quickly expressed and purified; and the method expands the secretion and expression path of the protein, can realize the simultaneous expression of a plurality of heterologous proteins, and can effectively improve the expression amount of the target protein.

The invention discloses an efficient preparation and application method of antibacterial peptide special for livestock and poultry, and belongs to the technical field of bioengineering and livestock and poultry breeding. Aiming at the problems that the existing antibacterial peptide preparation process is complicated, the yield is low, the targeting property is poor, the antibacterial peptide is easily degraded by gastrointestinal tracts of livestock and poultry in application, and the balance of intestinal flora is damaged, the pichia pastoris modified by genetic engineering is adopted as an expression host, and the efficient secretory expression of the antibacterial peptide is realized by optimizing a two-stage fermentation process and a culture medium formula. Complex purification steps are not needed, the yield of the antibacterial peptide reaches 85% or above, the activity retention rate is increased by 40%, the intestinal pathogenic bacteria of livestock and poultry can be specifically inhibited, no damage is caused to probiotics, the antibacterial peptide can be prepared into feed additives, oral preparations and the like to be applied to livestock and poultry breeding, antibiotic dependence is reduced, the breeding cost is reduced, and remarkable economic and ecological benefits are achieved.

The invention relates to the technical field of bioengineering and environmental biology, and particularly discloses high-temperature-resistant polylactic acid depolymerases and application thereof in degradation of polylactic acid plastics. The nucleotide sequence of the coding gene of the high-temperature-resistant polylactic acid depolymerases B01 is as shown in SEQ ID NO: 1, and the amino acid sequence of the coding gene is as shown in SEQ ID NO: 2. The enzyme is derived from metagenome mining, the sequence similarity of the enzyme and the known PLA depolymerases PAM is only 34.45%, and the enzyme has remarkable novelty. According to the present invention, the secretory expression and the purification of the B01 are successfully achieved in the bacillus subtilis SCK6; an enzymatic property research shows that the optimal reaction temperature of the B01 is 65 DEG C, the half-life period at 60 DEG C is as long as 150 minutes, and the B01 shows excellent thermal stability; and the optimum pH value of the strain is 9.0. The invention provides a novel enzyme preparation with important application potential for high-temperature biological recovery treatment of PLA plastic waste.

The present application relates to the field of molecular biology, in particular to a recombinant human heparin N-deacetylase encoding gene, a recombinant vector, a recombinant strain and application thereof, the recombinant human heparin N-deacetylase constructed in the expression process shows excellent stability and activity, has the ability of specific catalysis of deacetylation of acetyl group in heparin precursor, and can effectively generate deacetylated heparin precursor. This property significantly improves the conversion efficiency and reduces the generation of by-products. The present application optimizes the codon of the recombinant human heparin N-deacetylase, and uses the promoter P AOX1 or P GAP to regulate the expression, so that the expression level of the recombinant human heparin N-deacetylase in Pichia pastoris is significantly improved, and the obtained GS115-AOX1-hNDase and GS115-PGAP-hNDase strains successfully realize the secretory expression of the target gene, providing a new strategy and method for the biosynthesis of heparin.

The invention discloses a preparation method of pichia pastoris expression secreting type deglycosylation alkaline phosphatase, and belongs to the technical field of biological engineering. According to the invention, the GS115-BIAP II pichia pastoris genetically engineered bacterium capable of secreting and expressing bovine intestine alkaline phosphatase BIAP II is prepared by using a conventional method. The method comprises the following steps: carrying out site-directed knockout on an ALG3 gene for coding alpha-1, 3-mannosyl transferase and an OCH1 gene for coding alpha-1, 6-mannosyl transferase by using a CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats-associated Protein 9) technology to prepare a glycosylation gene defect type engineering bacterium GS115-delta ALG3-delta OCH1-BIAP II. The secretory expression BIAP II is deglycosylated alkaline phosphatase, and the specific activity of the secretory expression BIAP II is close to that of natural BIAP II. The deglycosylated alkaline phosphatase can improve the antibody coupling efficiency and marking uniformity under specific conditions. The method has remarkable advantages in chemiluminescence diagnosis, high-sensitivity immunodetection and development of multifunctional biological coupling reagents.

The present application relates to the field of biosynthesis and microbial fermentation engineering technology, in particular to a kind of secretion expression human lactoferrin Yarrowia lipolytica and its construction method and application.The present application provides a kind of secretion expression human lactoferrin recombinant Yarrowia lipolytica, the secretion expression of human lactoferrin in the recombinant Yarrowia lipolytica is guided by secretion signal peptide;The expression of chaperone HAC1, Cne and Pdi1 is simultaneously enhanced.The present application realizes the secretion expression of human lactoferrin in Yarrowia lipolytica, effectively promotes the folding and secretion expression of human lactoferrin by the cooperation of chaperone, significantly improves the yield of human lactoferrin, and has important application value in the industrial scale production of human lactoferrin.

The invention relates to the technical field of biology, in particular to a recombinant pichia pastoris engineering bacterium, a construction method of the recombinant pichia pastoris engineering bacterium and application of the recombinant pichia pastoris engineering bacterium in secretory expression of ovotransferrin. The heterologous expression of ovotransferrin is effectively realized by optimizing ovotransferrin genes and utilizing molecular chaperone genes to strengthen the regulation and control capability of hosts, and meanwhile, by optimizing signal peptides and constructing high-copy recombinant pichia pastoris expression strains, the low-copy expression condition is verified and compared by utilizing fermentation expression, so that the heterologous expression of ovotransferrin is realized. According to the high-copy recombinant pichia pastoris engineering bacteria constructed by the invention, the ovotransferrin expression quantity is obviously improved and reaches 140 mg / L, the secretory expression efficiency of the ovotransferrin is effectively improved, and theoretical guidance is provided for heterologous expression and industrial production of the ovotransferrin.

The invention discloses a method for secretory expression of recombinant mechanical structural protein by using escherichia coli and application, and relates to the field of bioengineering. According to the method, signal peptides are introduced to the N end of recombinant mechanical structural proteins, and the signal peptides, namely OmpA, PelB, PhoA, DsbA and STII, capable of guiding different recombinant mechanical structural proteins to secrete and express are screened from six natural signal peptides from escherichia coli and other species. Meanwhile, the yield of extracellular protein is further improved by combining an amino acid tRNA supplement strategy. According to the method, the recombinant escherichia coli strain capable of efficiently secreting and expressing the mechanical structure protein can be obtained, the downstream purification process is simplified, and the method has important application value.

The invention relates to the technical field of biology, in particular to a pagrosomus major antibacterial peptide Pm-NK-lysin as well as a preparation method and application of the pagrosomus major antibacterial peptide Pm-NK-lysin. The amino acid sequence of the antibacterial peptide is as shown in SEQ ID NO: 1. The minimum inhibitory concentration of the antibacterial peptide to vibrio anguillarum is not higher than 8 [mu] g / ml, and / or the minimum inhibitory concentration of the antibacterial peptide to escherichia coli is not higher than 4 [mu] g / ml. The antibacterial peptide is prepared through a pichia pastoris expression system. The antibacterial peptide is applied to preparation of an antibacterial preparation. According to the invention, efficient secretory expression of recombinant Pm-NK-lysin is successfully realized by creatively utilizing a pichia pastoris eukaryotic expression system, and a protein purification process is optimized, so that sufficient sources of high-activity recombinant protein are guaranteed. The Pm-NK-lysin provided by the invention has an excellent killing effect on aquatic pathogenic bacteria. The antibacterial peptide Pm-NK-lysin is used for replacing or reducing the use of antibiotics to control the harm of aquatic bacterial diseases, has wide popularization and application values, and certainly has far-reaching significance for promoting the green and healthy development of the aquaculture industry.

The invention belongs to the technical field of gene recombination, and particularly relates to a recombinant human III-type triple helix collagen, a recombinant yeast strain for expressing the recombinant human III-type triple helix collagen and a construction method of the recombinant human III-type triple helix collagen. The amino acid sequence of the recombinant human III-type triple-helix collagen is shown as SEQ ID NO. 1, and the recombinant human III-type triple-helix collagen has triple-helix conformation close to a natural biological structure of collagen and has good application value and potential. According to the recombinant yeast strain disclosed by the invention, human-derived III-type collagen and proline hydroxylase are co-expressed in yeast cells without YPS1 protease, and meanwhile, a gene beneficial to vesicle transport and protein folding is transferred, so that hydroxylation of the recombinant collagen can be realized, a natural triple helix conformation is formed, and the recombinant yeast strain can be used for preparing a recombinant collagen product. The degradation problem of the collagen in the expression process can be solved, and the extracellular secretion expression of the triple helix collagen is realized.

The application relates to a recombinant type I collagen and application thereof, which utilizes the characteristics of Pichia pastoris capable of secreting and expressing recombinant proteins, secretes the recombinant type I collagen on the culture medium supernatant, and reduces the difficulty of protein purification. In addition, the integrin site RGD is inserted into the collagen to form a recombinant type I collagen with high stability and high expression. According to the relative proliferation of cells and the relative adhesion experiment of cells, it can be seen that the recombinant type I collagen has the effects of promoting the proliferation of skin fibroblasts, improving the skin, promoting wound healing, promoting the adhesion of fibroblasts in the skin layer and the like. Therefore, the recombinant type I collagen has great potential applications in the fields of skin care and beauty, medical treatment and the like.

The application provides a method for detecting secretion expression of a target protein, which comprises the following steps: fusing the target protein with a fluorescence-enhanced nanobody to obtain a fusion protein; adding the obtained fusion protein into a system containing a fluorescent protein and a fluorescence-inhibited nanobody to perform a reaction; and measuring the fluorescence intensity of the system after the reaction is completed, and calculating the secretion expression amount of the target protein according to the change amount of the fluorescence intensity. In the application, the GBP1 is fused with the target protein, and the expression interference of the GBP1 on the target protein is reduced by adding a connecting peptide, so that the secretion expression amount of the target protein is determined. The method of the application can detect a wide concentration range of the target protein, has a high signal response multiple, and can be applied to a shake flask, a well plate and a droplet microfluidic screening system.

The invention provides a coding gene of canine serum albumin, an expression vector, a recombinant engineering bacterium, and a preparation method and application of the recombinant engineering bacterium of canine serum albumin, the coding gene of canine serum albumin comprises a co-translation translocation chimeric signal peptide sequence and a canine serum albumin mature peptide sequence, the co-translation translocation chimeric signal peptide sequence comprises an Ost1 signal peptide pre-peptide region and an alpha-factor signal peptide pro-peptide region of saccharomyces cerevisiae, and the Ost1 signal peptide pre-peptide region and the alpha-factor signal peptide pro-peptide region are chimeric to obtain the co-translation translocation chimeric signal peptide sequence. According to the coding gene of the dog serum albumin provided by the invention, the efficient secretory expression of the dog serum albumin in pichia pastoris cells is realized by utilizing the synergistic effect of an Ost1 signal peptide pre-peptide region and an alpha-factor signal peptide pro-peptide region in a co-translation translocation chimeric signal peptide sequence.

The invention relates to the technical field of gene engineering, in particular to high-activity peptide ligase and application thereof in synthesis of tilpotide. The high-activity peptide ligase provided by the invention shows catalytic efficiency far higher than that of an existing enzyme in a specific ligation reaction aiming at an acyl donor and an acyl receptor of a precursor fragment of the tilpoitide; by systematically transforming a host strain, the high-activity peptide ligase which can be secreted and expressed at high level and high fidelity and has correct folding and complete activity is obtained, and meanwhile, the degradation of a product is reduced to the maximum extent; and an optimal expression element combination of the high-activity peptide ligase is obtained through high-throughput optimization of an expression system. The method is finally integrated into a complete new process for synthesizing the tilpoitide by the chemical enzyme method, which can realize kilogram-level production, and is suitable for popularization and application.

The application discloses a method for increasing 2-pyrrolidone synthesis and belongs to the technical field of bioengineering. tacm The CoA transferase from Butyricicoccus faecihominis is expressed in the coryneform bacterium glutamicum, and the CoA transferase is secreted and expressed by means of P The promoter and the NS signal peptide strengthen the secretory expression of the CoA transferase, so that the constructed strain can ferment 2-pyrrolidone in a glucose carbon source and in a pH 7.0 environment, and the yield of the 2-pyrrolidone can reach 12.13 g / L in 96 h of flask fermentation.

The invention belongs to the technical field of biology, and provides a signal peptide for improving the secretory expression of a recombinant protein of a mammalian cell and application of the signal peptide, the structure of the signal peptide for improving the secretory expression of the recombinant protein of the mammalian cell is 5 '-[IgG light chain N terminal 15aa]-[albumin hydrophobic core 8aa]-[basic amino acid enrichment region]-3', and the specific sequence is SEQ ID NO.1 MDFQVQIFSFLLISASVIMSRKRKR. The signal peptide provided by the invention secretes and expresses a target protein in mammalian cells, has universality, can remarkably improve the secretion efficiency, can realize efficient secretion and expression of various recombinant proteins, and has very important significance.

The invention provides glycoprotein phosphoglycerate kinase derived from stropharia rugoso-annulata, a recombinant vector, recombinant pichia pastoris engineering bacteria and application of the glycoprotein phosphoglycerate kinase, and belongs to the technical field of phosphoglycerate kinase production. The phosphoglycerate kinase S.rug.-PGK sourced from S.rug.-PGK provided by the invention has an amino acid sequence as shown in SEQ ID NO. 1, and a nucleotide sequence for coding the S.rug.-PGK is as shown in SEQ ID NO. 2. According to the invention, novel phosphoglycerate kinase (S.rug.-PGK) derived from Stropharia rugoso-annulata is taken as a target, and efficient secretion production of S.rug.-PGK is realized by systematically optimizing a pichia pastoris expression platform. According to the invention, efficient secretory expression of S.rug.-PGK in pichia pastoris is realized for the first time, it is clear that the pichia pastoris X33 strain is an optimal production system under the conditions that OD600 is equal to 2 and the inducer concentration is 1%, and a foundation is laid for large-scale preparation.

The invention provides a transgenic expression cassette, a recombinant eukaryotic expression vector, a eukaryotic cell, an NS1 protein and a preparation method and application thereof, and relates to the technical field of biology. The transgenic expression cassette provided by the invention is matched with an eukaryotic expression system, efficient secretory expression of subtypes in NS1 protein 4 is successfully realized, the obtained NS1 protein retains native conformation and glycosylation modification, the antigen specificity is obviously superior to that of a prokaryotic expression product, and the transgenic expression cassette can be used for preparation of an NS1 antibody and research and development of an NS1 protein detection product.

This invention relates to bioengineering, specifically disclosing a method for preparing recombinant humanized type III collagen with a triple-helix structure, comprising the following steps: S1, constructing a ppICZαA recombinant expression plasmid containing a human type III collagen fragment, and transforming the recombinant expression plasmid into yeast; S2, screening high-copy transformant strains from the strain and performing shake-flask level induction expression tests; S3, conducting high-density fermentation in a fermenter to obtain a large amount of secretible target protein. This invention, by constructing a ppICZαA recombinant expression plasmid containing a human type III collagen fragment and electroporating it into a yeast expression system, screens for yeast strains capable of expressing recombinant humanized type III collagen fragments, and conducts high-density fermentation in a fermenter, followed by purification after fermentation, yields pure recombinant collagen with a triple-helix structure, which can be applied in skincare products, functional skincare products, medical devices, and biomedical materials.

The invention belongs to the fields of bioengineering and animal nutrition, and relates to recombinant lactobacillus plantarum for secretory expression of chicken IGF-1 (Insulin-like Growth Factor-1) as well as a construction method and application thereof. The recombinant lactobacillus plantarum takes lactobacillus plantarum as a host, the host contains a recombinant expression vector, and the recombinant expression vector contains an SlpA promoter, a lactobacillus plantarum extracellular protein Lp2007 signal peptide gene and a chicken IGF-1 gene; the sequence of the chicken IGF-1 gene is as shown in SEQ ID NO: 1, the sequence of the SlpA promoter is as shown in SEQ ID NO: 2, and the sequence of the lactobacillus plantarum extracellular protein Lp2007 signal peptide is as shown in SEQ ID NO: 3. The recombinant lactobacillus plantarum can secrete IGF-1 outside cells of the recombinant lactobacillus plantarum. When the recombinant lactobacillus plantarum for secreting and expressing the chicken IGF-1 is used for feeding chickens, the growth speed of the chickens can be remarkably increased, cost reduction and efficiency improvement of the poultry raising industry are realized, and the recombinant lactobacillus plantarum has important economic and social values.

The invention discloses a method for improving the expression quantity and immunogenicity of a novel plant-derived coronavirus RBD-sc-dimer, and claims to protect a recombinant plasmid, and the recombinant plasmid comprises an optimized target protein sequence. The optimized target protein sequence sequentially comprises a gene sequence of a signal peptide as shown in a sequence 2, a gene sequence of a target protein dimer and a nucleotide sequence as shown in a sequence 8 from the 5'end to the 3 'end; the optimized target protein sequence is positioned behind the 35S promoter in the plasmid. According to the plant source protein prepared through the recombinant plasmid, RBD-sc-dimer is fused with 10 serine-proline dipeptides, and secretory expression is achieved through an SSExt signal peptide. Experimental results show that by means of the design, the expression quantity of RBD-sc-dimer can be increased by 2.4 times, and the total IgG level after three times of immunization of 10 g of recombinant protein is increased by about 100 times.

The present application relates to a kind of mediated zymolytic enzyme of brown algae and application of double transcription unit recombinant bacteria of oxyprotein and brown algae, belong to genetic engineering technical field, the recombinant engineering bacteria is composed of two transcription units, one transcription unit is GAP-alpha-Vgb-AOX1TT, another transcription unit is AOX-alpha-102C300C-AOX1TT, two transcription units are expressed in tandem in pPICZ alphaA plasmid.The present application also provides the preparation method and application of the recombinant bacteria.The recombinant bacteria of the present application can improve the secretion expression efficiency of brown algae zymolytic enzyme, especially can be expressed in low-oxygen environment.

The invention relates to an mRNA nucleic acid molecule of an avian reticuloendotheliosis virus and application, and belongs to the technical field of immunology and biologica.According to the nucleic acid molecule, a T7 promoter, a 5 '-untranslated region (5' UTR), a gp90 antigen coding gene, a 3 '-untranslated region and poly (adenosine monophosphate) are used for constructing a recombinant plasmid. According to the invention, a gp90 gene sequence and polyadenylic acid are optimized to obtain the mRNA nucleic acid molecule of the reticuloendotheliosis of poultry, and the mRNA nucleic acid molecule is prepared into the mRNA vaccine; the constructed candidate mRNA vaccine is expressed in a 293T cell and is subjected to extracellular secretory expression.

The invention relates to the field of genetic engineering and fermentation engineering, and particularly discloses a method for exogenous expression of COQ3 by recombinant pichia pastoris engineering bacteria. According to the invention, a target gene COQ2 is inserted into a pichia pastoris secretory expression vector ppicZA to construct a recombinant expression plasmid. The plasmid is transformed into a pichia pastoris GS115 host bacterium to obtain a recombinant pichia pastoris engineering bacterium capable of expressing COQ3. Through methanol-induced fermentation, the engineering bacterium can accumulate COQ3 in cells. The method provided by the invention has the advantages of low cost, easiness in large-scale production and the like, and provides a new way for realizing industrial production of COQ3 and other ubiquinone substances.

The application discloses a recombinant membrane-targeting type 17 collagen protein and application thereof. The application realizes gene fusion of a COL17A1 key function domain and an independent membrane-targeting structure domain for the first time, constructs a novel "activity-targeting" integrated recombinant membrane-targeting type 17 collagen protein (M-C17a1), effectively integrates function activity and cell targeting property through ingenious structure design, and exhibits excellent performance in resisting skin aging, tissue repair and promoting hair growth. The application also proposes a complete preparation method based on a Pichia pastoris expression system. The expression system has high-density fermentation capacity and secretory expression characteristics, significantly simplifies a purification process, and has relatively low production cost, so that the expression system is suitable for large-scale production of the recombinant protein and has good industrial application prospect.

The invention relates to a method for improving the secretory expression level of expansin in bacillus subtilis, and aims to provide a method capable of promoting the bacillus subtilis to secrete expansin (microorganism-derived expansin BsEXLX1 and plant-derived expansin LeEXP2). The method comprises the following steps: firstly, screening a promoter capable of efficiently starting expansin to construct an engineering strain, and then performing overexpression of a secretory cofactor in the strain to improve the limitation of target protein in the aspects of secretory transport, folding maturation, stability and the like, thereby improving the extracellular secretion level of the two expansin. Amplification verification is carried out through a 5L fermentation tank, the secretion level of expansin is further improved, the expression quantity of BsEXLX1 protein is improved to 1.3 g / L, and the secretion quantity of LeEXP2 is improved to 43.7 mg / L. And a process basis and industrialization potential are provided for subsequent large-scale fermentation production and application development.