289 results about "Pichia pastoris" patented technology

The invention relates to a recombinant pichia pastoris strain capable of producing astaxanthin at high yield as well as a construction method and application of the recombinant pichia pastoris strain. The recombinant pichia pastoris strain is obtained by expressing ATP (adenosine triphosphate) citrate lyase (ACL) and acetyl-CoA synthase (ACS) or phosphoketolase (PK) and phosphotransacetylase (PTA), choline kinase (CK), inositol polyphosphate kinase (IPK) and vitreoscilla hemoglobin (VHB) in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified strain PP-LC2. The astaxanthin production performance of the recombinant strain is verified on the basis of comparison of precursor supply pathways, IUP pathway construction and improvement of the oxygen supply capacity of the engineering strain, and the astaxanthin production capacity of the pichia pastoris is further improved. The construction method of the recombinant pichia pastoris is simple, the synthesis of astaxanthin can be better promoted, the engineering strain can efficiently synthesize the astaxanthin by utilizing methanol through amplification fermentation of a 5L fermentation tank, and industrial production is facilitated.

The invention provides a micromolecular recombinant XVII type collagen as well as a preparation method and application thereof, and relates to the technical field of protein engineering. The novel recombinant human XVII type collagen is obtained by optimizing and screening a plurality of spiral region sequences such as a collagen structural domain, a C-terminal region and a middle region of the human XVII type collagen. The amino acid composition of the recombinant XVII type collagen is consistent with that of a natural collagen alpha1 chain, immunological rejection is not generated when the recombinant XVII type collagen is applied to a human body, the recombinant XVII type collagen can be widely applied to surgical medical treatment and medical beauty industries, the molecular weight of the collagen is small, efficient secretion soluble expression in eukaryotic host cells such as pichia pastoris can be achieved, and the recombinant XVII type collagen has a good application prospect. And industrial large-scale production is carried out. According to the present invention, the verification results show that the micromolecular recombinant XVII type collagen further has basement membrane anti-aging activity, wherein the basement membrane anti-aging activity specifically comprises the improvement of the expression levels of the IV type collagen, the VII type collagen and the XVII type collagen;

The embodiment of the invention provides a recombinant pichia pastoris strain as well as a construction method and application thereof. The construction method comprises the following steps: by taking a pichia pastoris strain as a host strain, constructing a dual-synthesis path of phenylalanine ammonialyase and tyrosine ammonialyase in the host strain to obtain a first engineering strain; removing chorismic acid feedback inhibition of the first engineering strain based on metabolic engineering to obtain a second engineering strain; optimizing an aromatic amino acid synthesis route of the second engineering strain to obtain a third engineering strain; knocking out a branch metabolic pathway of the third engineering strain to obtain a fourth engineering strain; increasing precursor supply of the fourth engineering strain to obtain a recombinant pichia pastoris strain; according to the construction method provided by the embodiment of the invention, the synthesis path of p-coumaric acid can be systematically optimized, the recombinant pichia pastoris strain for efficiently producing p-coumaric acid by taking methanol as a carbon source is constructed, and compared with a host strain, the recombinant pichia pastoris strain has the advantage that the yield of p-coumaric acid is remarkably increased.

The invention relates to the technical field of microorganisms, and provides pichia pastoris CMRC 13Y with high protein yield and application of the pichia pastoris CMRC 13Y. The pichia pastoris CMRC 13Y is separated from naturally-fermented dried milk cakes, the classification name of the pichia pastoris CMRC 13Y is Pichia kudriavzevii, and the preservation number of the pichia pastoris CMRC 13Y is CGMCC No.34655. The pichia pastoris CMRC 13Y has high acid resistance, and the growth of the pichia pastoris CMRC 13Y is not obviously influenced along with the reduction of the pH value within the pH value range of 3.0-7.0; a yeast protein product prepared from the strain CMRC 13Y is subjected to sensory flavor determination, has no earthy smell in the aspect of smell, has fragrance, is fine and smooth in mouth feel, and has no unpleasant taste; the strain can achieve high-biomass growth and high-protein expression, has the advantages of being green, environmentally friendly, high in protein yield and the like, can be used for preparing microbial protein related products and is wide in application prospect.

The invention provides a pichia pastoris strain for efficiently converting methanol to produce high-methionine-content single-cell protein and application of the pichia pastoris strain, and belongs to the technical field of biology. According to the invention, a strain of pichia pastoris (named as Pichiapastoris CN-SCP 00 with a preservation number of CCTCC (China Center For Type Culture Collection) NO: M2025657) is obtained through separation, and is subjected to multiple rounds of ARTP iterative mutation breeding, so that a strain Pichiapastoris CN-SCPYE421 (with a preservation number of CCTCC NO: M2025658) for efficiently converting methanol to produce single-cell protein containing high methionine is obtained. Compared with a commercial strain, the Pichiapastoris CN-SCPYE421 has higher methanol tolerance and utilization capacity, the fermentation time is remarkably shortened, and in addition, the Pichiapastoris CN-SCPYE421 can well grow at the temperature of 35 DEG C. The Pichiapastoris CN-SCPYE421 is subjected to fermentation culture in a 5L fermentation tank, the dry weight of thalli reaches 120-150g / L, the protein content reaches 65-75% of the dry weight of the thalli, the Pichiapastoris CN-SCPYE421 is rich in various essential amino acids, and the methionine content reaches 2.3%. The invention provides an excellent strain for industrial development of single-cell protein, and has great social significance and economic value for the feed protein industry.

The invention discloses a recombinant pichia pastoris strain with high yield of retinaldehyde as well as a construction method and application of the recombinant pichia pastoris strain. The recombinant pichia pastoris strain is obtained by expressing dehydrogenase Zwf1, phosphate dehydrogenase Gnd2, NADH kinase Pos5 and two copied beta-carotene-15, 15 '-monooxygenase BCMO in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified strain PP-B (Propene Polymer-B). The retinaldehyde production performance of the recombinant strain is verified based on cofactor engineering and the copy number of key enzymes, and the retinaldehyde production capacity of the pichia pastoris is further improved. The recombinant pichia pastoris disclosed by the invention is simple in construction method, can better promote the synthesis of retinol, and is beneficial to industrial production as the engineering strain can efficiently synthesize retinol by using methanol as a unique carbon source through amplification fermentation in a 5L fermentation tank.

The invention relates to pichia pastoris engineering bacteria and a preparation method thereof, and a method for producing cellooligosaccharide / disaccharide by using corn straws or wheat straws, and belongs to the technical field of genetic engineering. A cellulose incision enzyme gene and a cellulose excision enzyme gene are transferred into pichia pastoris engineering bacteria. The preparation method comprises the following steps: respectively inserting a cellulose incision enzyme gene and a cellulose excision enzyme gene into plasmids to construct expression vectors; transferring the expression vector into a competent cell of pichia pastoris GS115; and screening and identifying to obtain successfully transformed pichia pastoris engineering bacteria. According to the present invention, the pichia pastoris heterologous expression cellulose incision enzyme and cellulose excision enzyme are adopted to construct the pichia pastoris engineering strain capable of degrading the plant straw into the cellooligosaccharide / disaccharide, such that the reliable chassis strain is provided for the production of the starch from the cellulose;

The application provides a recombinant Pichia pastoris strain and a construction method and application thereof. The construction method comprises the following steps: taking a Pichia pastoris strain as a host strain, constructing a double synthesis pathway of phenylalanine ammonia lyase and tyrosine ammonia lyase in the host strain to obtain a first engineering strain; relieving the branch acid feedback inhibition of the first engineering strain based on metabolic engineering to obtain a second engineering strain; optimizing the aromatic amino acid synthesis pathway of the second engineering strain to obtain a third engineering strain; knocking out the branch metabolic pathway of the third engineering strain to obtain a fourth engineering strain; increasing the precursor supply of the fourth engineering strain to obtain the recombinant Pichia pastoris strain. The construction method can systematically optimize the synthesis path of p-coumaric acid, and the recombinant Pichia pastoris strain for efficiently producing p-coumaric acid with methanol as a carbon source is constructed. Compared with the host strain, the yield of p-coumaric acid of the recombinant Pichia pastoris strain is significantly improved.

The invention relates to a method for producing vanillin from natural lignocellulose biomass through co-culture of pichia pastoris, which comprises the following steps: respectively constructing a xylan hydrolysis recombinant strain (for releasing ferulic acid) and a vanillin synthesis recombinant strain (for converting ferulic acid) to realize conversion production from a natural xylan substrate to vanillin. In order to reduce the generation of by-products, 15 genes for coding aldehyde dehydrogenase (ALDRs), alcohol dehydrogenase (ADHs) and aldehyde ketoreductase (AKRs) are further knocked out from the vanillin synthesis recombinant strain, so that the vanillin can be further converted into the by-product vanillic acid, thereby improving the yield and purity of the vanillin.

The invention discloses a recombinant engineering bacterium for expressing fetuB and application of the recombinant engineering bacterium, and relates to the technical field of genetic engineering. According to the recombinant engineering bacteria, pichia pastoris serves as host bacteria, after fetuB genes are introduced, efficient expression of the fetuB in the pichia pastoris is achieved by optimizing codons and improving fermentation conditions, and the expression quantities of CBS7435 under the shake-flask culture condition are 320 + / -25 mg / L, SMD116 under the shake-flask culture condition is 280 + / -30 mg / L, and SuperMan5 under the shake-flask culture condition is 580 + / -45 mg / L. After fermentation conditions are optimized, the culture yield of a fermentation tank can reach 27.35 g / L. The fetuin B disclosed by the invention is good in biological activity, high in expression, simple in operation process and suitable for large-scale industrial production.

The invention discloses a recombinant pichia pastoris strain as well as a construction method and application thereof, and belongs to the field of microbial fermentation. The construction method specifically comprises the following steps that pGAPZB is used as a plasmid skeleton, a recombinant expression plasmid pGAPZB-ACSec containing an ACSec gene is constructed, and the sequence of the ACSec gene is shown as SEQ ID No.1; linearizing the recombinant expression plasmid, and transforming the linearized recombinant expression plasmid into a pichia pastoris host; and screening a pichia pastoris strain capable of expressing the ACSec gene. According to the invention, the ability of pichia pastoris to grow and metabolize by utilizing acetic acid is improved, and the application potential of pichia pastoris in fermentation production and synthesis of chemical products taking acetic acid as a precursor by utilizing a cheap carbon source is shown.

The invention discloses pichia kudriavzevii for high yield of L-malic acid as well as a construction method and application thereof, and belongs to the technical field of biology, the pichia kudriavzevii is named as YM-023 and classified as Pichia kudriavzevii, the preservation number is CGMCC No. 35487, the preservation time is July 31, 2025, and the preservation number is CGMCC No. 35487. The preservation unit is China General Microbiological Culture Collection Center (CGMCC). The method comprises the following steps: starting from an acid-resistant yeast Kudriavzevii pichia pastoris strain YL-000 (the preservation number is CGMCC No.35680), carrying out metabolic engineering transformation to obtain a transformed strain, and ensuring that the transformed strain can realize efficient production of L-malic acid in a fermentation mode of adding a small amount of calcium carbonate neutralizer.

The invention relates to recombinant pichia pastoris capable of efficiently expressing III-type human-like collagen and application of the recombinant pichia pastoris, and belongs to the technical field of microorganisms. According to the invention, an III-type human-like collagen expression cassette is constructed in pichia pastoris GS115 and co-expressed with a molecular chaperone, a transcription factor and a translation factor, so that the expression efficiency of the III-type human-like collagen is improved, and the shake-flask fermentation yield reaches 0.93 g / L. And then fermentation conditions, material supplementing conditions, methanol feeding induction conditions and the like are optimized, so that the yield of the III-type human-like collagen in a 5L fermentation tank by the constructed recombinant pichia pastoris reaches 10.3 g / L, the efficient production of target protein is realized, a favorable tool is provided for the application of the III-type human-like collagen in the fields of food, cosmetics, medicines and the like, and the engineering application prospect is broad. Good industrial application prospects are realized.

The invention relates to pichia pastoris for producing brazitame protein as well as a construction method and application of the pichia pastoris, and belongs to the technical field of microorganisms. ExpL SP is used as a signal peptide in pichia pastoris, heterologous expression of Brazilian sweet protein is carried out through an AOX1 promoter, the ExpL SP and a Pro region of an alpha mating factor are further fused and transformed to obtain a new signal peptide to enhance the secretion efficiency, and the new signal peptide and a ribosomal protein synthesis factor Bcy1 are co-expressed to improve the protein synthesis capacity. The constructed pichia pastoris can be used for efficiently secreting the Brazilian sweet protein, the problems that in the prior art, when microorganisms are used for fermenting the Brazilian sweet protein, the Brazilian sweet protein is mistakenly folded, the sweet taste is reduced or lost are solved, the shake flask yield is stabilized at 450 mg / L, the fermentation tank yield reaches 5.215 g / L, and a high-cost-performance scheme is provided for industrial production of the Brazilian sweet protein.

The invention discloses a method for efficiently expressing FAD-dependent glucose dehydrogenase in pichia pastoris. The method comprises the following steps: (1) constructing a recombinant plasmid containing a promoter, a signal peptide, a glucose dehydrogenase gene and a purification tag; the nucleotide sequence of the signal peptide is as shown in SEQ ID No.2; (2) transforming the recombinant plasmids into host bacteria pichia pastoris to obtain recombinant pichia pastoris engineering bacteria capable of expressing FAD-dependent glucose dehydrogenase; and (3) carrying out fermentation culture, separation and purification on the recombinant pichia pastoris engineering bacteria to obtain the FAD-dependent glucose dehydrogenase. According to the invention, the expression level and the final yield of the FAD-GDH are obviously improved.

The present invention belongs to the field of microbial metabolic engineering and synthetic biology technology applications, and particularly relates to a Pichia pastoris engineered strain that produces high sclareol production, as well as its construction method and application. A sclareol biosynthesis pathway is constructed in a host strain, and the intracellular mevalonate metabolic pathway and the central metabolic pathway are optimized, thereby obtaining an engineered bacterial strain A; the host strain is Pichia pastoris; or, metabolic regulatory factors are overexpressed or knocked out in the above-obtained engineered bacterial strain A, thereby obtaining an engineered bacterial strain B; or, in the above-obtained engineered bacterial strain B, a cell compartmentalization strategy is used to target the synthesis pathway to the peroxisome and optimize it, thereby obtaining an engineered bacterial strain C. The present invention provides a Pichia pastoris chassis cell for synthesizing the diterpenoid compound sclareol and an engineered strain for synthesizing sclareol; by introducing and optimizing the expression of the sclareol synthesis pathway, the sclareol yields in shake flask batch fermentation and bioreactor batch fed-batch fermentation reach 631.6 mg / L and 10.5 g / L, respectively.

The invention relates to the technical field of biology, in particular to a pichia pastoris recombinant strain for expressing non-specific peroxygenase CciUPO and application of the pichia pastoris recombinant strain. The pichia pastoris recombinant strain is obtained by the following steps: S1, synthesizing a sequence of a CciUPO original gene sequence optimized based on a yeast codon; s2, carrying out recombination with the EV signal peptide after the directed evolution; s3, connecting the recombined target gene with an expression vector to construct a recombinant plasmid; and S4, transforming the recombinant plasmid into pichia pastoris competent cells, and screening out a recombinant strain containing the CciUPO recombinant gene. The expression quantity of the CciUPO obtained through the operation is increased by 10 times compared with that of a wild type CciUPO, the CciUPO has strict catalytic regioselectivity, can catalyze vitamin D3 to synthesize calcifediol, the selectivity is as high as 95%, the catalytic conversion rate of toluene is 95%, the selectivity is 78%, and the CciUPO has a relatively high application prospect in industrial production.

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

The invention discloses an optimized recombinant chicken double-cell factor as well as a preparation method and application thereof. The chicken recombinant double-cell factor coding gene provided by the invention contains complete ChIL-2 and ChIFN-alpha structures. The preparation process comprises the following steps: selecting a pichia pastoris expression system; optimizing the recombinant chicken double-cell factor according to the codon preference of pichia pastoris; inserting the optimized gene sequence into an expression vector, and introducing host bacteria to construct a recombinant yeast engineering strain; carrying out inoculated culture by using the obtained recombinant yeast, and screening strains containing target protein based on expression level to obtain required culture bacteria; and breeding and culturing the obtained high-expression-level culture bacteria. According to the invention, ChIL-2 and ChIFN-alpha are connected through a modified flexible linker, and a nucleotide sequence is replaced by preferred codons of pichia pastoris, so that a chicken recombinant cell factor can be secreted to a supernatant, the defect of ChIL-2 expression quantity is overcome, and the problem that proteins of ChIL-2 and ChIFN-alpha mainly exist in bacteria and inclusion bodies in prokaryotic expression is solved.

The invention provides pichia pastoris capable of efficiently expressing TFF3 fusion protein as well as a construction method and application of the pichia pastoris, and belongs to the technical field of engineering bacteria. The preservation number of the pichia pastoris is CCTCC (China Center For Type Culture Collection) NO: M 2025013. The pichia pastoris constructed by the invention can obviously improve the expression quantity of the TFF3, the shake flask expression quantity is up to 391 mg / L, and the pichia pastoris has a wide application prospect in large-scale industrial production of the TFF3.

The invention discloses a non-specific peroxidase HecUPO derived from Hypoxylon sp. (strain EC38), wherein the amino acid sequence of the non-specific peroxidase HecUPO is as shown in SEQ ID NO: 1. The enzyme can efficiently catalyze testosterone to generate 16 alpha-hydroxylation reaction to generate C16 alpha-hydroxytestosterone, the conversion rate reaches 70%, the separation yield reaches 82%, no by-product is generated, and the C16 alpha-hydroxytestosterone has excellent regioselectivity. The invention also discloses a gene for coding the HecUPO, a recombinant plasmid pPICZA-HecUPO containing the gene, a recombinant pichia pastoris engineering bacterium and an application of the recombinant pichia pastoris engineering bacterium in steroid hydroxylation. The non-specific peroxidase HecUPO disclosed by the invention has the advantages of being high in catalytic efficiency, specific in regioselectivity, simple in reaction system, green, environmentally friendly and the like, and a new enzymology resource is provided for fixed-point hydroxylation modification of steroid drugs.

The invention provides an application of GMI of Ganoderma microspore in preparation of drugs and food, and the drugs and food are prepared from Ganoderma microspore globulin produced by fermentation of a genetically modified pichia pastoris Ey72 strain and used for treating, preventing or improving pulmonary fibrosis.

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

The invention relates to the technical field of biology, and particularly discloses pichia pastoris as well as a construction method and application thereof. According to the pichia pastoris disclosed by the invention, beta-glucan is knocked out to synthesize a related gene gas1. By deleting the beta-glucan coding gene gas1 gene of the pichia pastoris cell wall, the cell size is reduced, the transmission distance from the cell surface to the cell nucleus is reduced, oxygen transmission and product secretion during high-density culture are improved, and the fermentation production efficiency is improved. In addition, the pichia pastoris constructed by the invention enhances the synthesis of the intracellular product bisabolol, and also can enhance the secretion ability of the intracellular product bisabolol. By reducing the cell size, reducing the carbon flow flowing to the cell wall polysaccharide synthesis and increasing the carbon source flowing to the target product synthesis, the application has great significance in industrial production.

The invention discloses a potato protein with good emulsibility, gelling property and foamability, and belongs to the fields of biology and genetic engineering. According to the invention, the designed potato protein realizes recombinant expression in pichia pastoris, and it is verified that the recombinant expressed protein can form a secondary structure and can form hydrogel at a concentration of 60 g / L. The invention also provides an application of the prepared potato protein as a food additive, shows emulsifying and gelling capabilities superior to those of commercial potato protein, and has huge value and potential in the fields of food and tissue engineering.

The invention provides a strain of pichia pastoris SIVE9110 and an application of the pichia pastoris SIVE9110 in Morisseria wine. The strain SIVE9110 is preserved in the China Center for Type Culture Collection (CCTCC for short) on September 17, 2025; the address is Wuhan University, Wuchang District, Wuhan City, Hubei Province; the postal code is 430072, and the preservation number is CCTCC (China Center For Type Culture Collection) NO: M 20252052; the Latin name is Pichia occidentalis, and the Latin name is Pichia occidentalis; the strain SIVE9110 and saccharomyces cerevisiae are sequentially fermented, so that the wind and soil characteristics of the maisseria wine can be highlighted; and the prepared maisseria wine is softer and mellow in mouth feel.

The invention discloses a preparation method and application of hirudo hyaluronidase expressed by pichia pastoris, and belongs to the technical field of bioengineering.The preparation method comprises the steps of fermentation, pretreatment, one-step chromatographic purification and filtration. The pretreatment comprises the following steps: mixing and stirring a hirudo hyaluronidase fermentation solution and calcium chloride, adjusting the pH value to 7.2-7.6, centrifuging, collecting a supernatant, carrying out suction filtration by a filter membrane, taking a filtrate, filtering by a primary ultrafiltration membrane, and taking a permeate as a loading solution; the one-step chromatographic purification comprises the step of carrying out one-step chromatographic purification on the loading solution to obtain a chromatographic purification solution; according to the method disclosed by the invention, the hyaluronidase with high enzyme activity can be obtained, and the purified hyaluronidase with high enzyme activity can be used for carrying out enzymolysis on macromolecular sodium hyaluronate to prepare low-molecular-weight and oligomeric sodium hyaluronate.

The invention specifically discloses a fermentation process suitable for III-type collagen expression, and relates to the technical field of biological fermentation of recombinant genetic engineering bacteria. According to the method disclosed by the invention, a double-carbon-source mixed feeding strategy of alanine and methanol is used, so that the problems that when pichia pastoris expresses exogenous target protein by taking methanol as a unique carbon source and energy source, cell growth and protein expression fight for the carbon source and the energy source together, the metabolic burden of cells is increased, and the expression efficiency is low are solved; through a three-step methanol fed-batch strategy, the expression efficiency is improved, and the toxic action of methanol on cells is reduced, so that the yield of the III-type collagen is further improved. The fermentation process provided by the invention solves the problem of reducing the production cost while improving the high-efficiency and high-quality expression of the target product under the condition of high-density thallus fermentation.

The present application belongs to the field of microbial metabolic engineering and synthetic biology technology application, in particular to a high methanol tolerance yeast chassis strain, a high 3-hydroxypropionic acid yield engineering bacterium and application. The high methanol tolerance yeast chassis strain takes wild type Pichia pastoris or fatty acid producing Pichia pastoris as a starting strain, is modified according to at least one of the following conditions (a)-(h), and a high methanol tolerance strain is obtained, wherein (a) MIOX gene knockout; (b) DMA2 gene knockout; (c) MET17 gene knockout; (d) BUB2 gene knockout; (e) CLB4 gene knockout; (f) UBP12 gene knockout; (g) ZFP gene knockout; (h) AOX1 gene mutation. The high methanol tolerance yeast chassis strain is further modified to obtain a high 3-hydroxypropionic acid yield engineering bacterium. The present application combines rational and non-rational metabolic engineering strategies, realizes efficient biosynthesis of fatty acids and 3-hydroxypropionic acid with methanol as the only carbon source, and lays an important foundation for low-carbon green biological manufacturing of chemicals.

The present invention relates to the field of biological fermentation. Specifically, it relates to a method and application for fermenting Platycladus orientalis leaves with a composite bacterium, and the composite bacterium includes Lactococcus lactis and Pichia pastoris. The method includes the following steps: (1) pretreatment of Platycladus orientalis leaves; (2) enzymatic treatment of Platycladus orientalis leaves; (3) fermentation of Platycladus orientalis leaves; (4) treatment of the fermentation broth. The present invention has the following beneficial effects: (1) The present invention optimizes the enzymatic hydrolysis parameters, fermentation parameters, and the method for extracting the fermentation broth, and obtains a better fermentation method; (2) The fermentation of Platycladus orientalis leaves in the present invention simplifies the components compared with the fermentation of various traditional Chinese medicine raw materials in the prior art, making the invention more convenient; (3) The mouse experiments of the present invention prove that by using the optimized fermentation method, more flavonoids can be obtained, the anti-hair loss effect is better, and it is more beneficial to the application of Platycladus orientalis leaves.