31 results about "Geranyl pyrophosphate" patented technology

The invention provides a method for biosynthesizing menadione by using yarrowia lipolytica. The method is realized by constructing recombinant yarrowia lipolytica, and the construction method of the recombinant yarrowia lipolytica for synthesizing menadione comprises the following steps: intensively expressing a mevalonic acid pathway in the yarrowia lipolytica, and synthesizing menadione by introducing a synthesis pathway of heterologous geranyl and geranyl pyrophosphate. A sufficient isoprene side chain donor is provided for biosynthesis of menadione; on the basis, an aromatic isopentenyl transferase coding gene which is optimized by codons and has menadione substrate specificity is over-expressed in yarrowia lipolytica. The construction method of the yarrowia lipolytica for biosynthesis of menadione provided by the invention is simple to operate, the constructed yarrowia lipolytica can obtain a menadione product with relatively high concentration by taking commercially available menadione as a substrate, the biosynthesis steps are relatively short, and the yarrowia lipolytica has relatively high production and application values.

The invention provides a metabolic engineering transformation method of high-yield coenzyme Q10 of rhodobacter sphaeroides and application of transformed engineering bacteria. The method comprises the following steps: knocking out geranyl and geranyl pyrophosphate synthase gene crtE participating in a carotenoid synthesis route; performing overexpression on a farnesyl pyrophosphate synthase gene ispA; a histidine kinase coding gene prrB is mutated, and the mutant is introduced. According to the invention, a combined regulation strategy suitable for targeted regulation is found, the metabolic flux synthesized by the coenzyme Q10 is accurately optimized, and the capability of producing the coenzyme Q10 by the rhodobacter sphaeroides is greatly improved.

The application discloses recombinant escherichia coli for producing isopulegol and a construction method, and the construction method is as follows: a plasmid 1 is obtained by replacing an expression module of pETDuet-1 with an EM nucleotide fragment; a plasmid 2 is obtained by replacing an expression module of pRSFDuet-1 with the EM nucleotide fragment; an optimized geraniol synthase gene ObGES is integrated into the plasmid 1, and an optimized geranylgeranyl diphosphate synthase gene AgGPPS is integrated into the plasmid 1 to obtain a plasmid 3; an optimized EcGeDH is integrated into the plasmid 3 to obtain a plasmid 4; an optimized SsOYE2.6 W78Y / C113I is integrated into the plasmid 2, and an optimized ZmSHC F486C is integrated into the plasmid 2 to obtain a plasmid 5; the plasmids 4 and 5 are introduced into escherichia coli to obtain the recombinant escherichia coli for producing isopulegol; and the recombinant escherichia coli of the application can obtain isopulegol after fermentation.

The invention provides a method for detecting geranyl pyrophosphoric acid in water by headspace solid-phase microextraction / gas chromatography-mass spectrometry, which utilizes the characteristic that geranyl pyrophosphoric acid is easy to decompose into linalool under an acidic condition, adsorbs linalool by headspace solid-phase microextraction, and indirectly quantifies geranyl pyrophosphoric acid in combination with a gas chromatography-mass spectrometry technology. The method is easy to operate, low in instrument cost, high in sensitivity and suitable for detecting geranyl pyrophosphoric acid in water bodies such as source water, the generation risk of 2-methylisoborneol can be pre-judged through the content of geranyl pyrophosphoric acid, and technical support is provided for water supply safety.

The invention belongs to but is not limited to the technical field of plant genetic engineering, and discloses a Nerolidol / Linalool synthetase gene MnTPS6 of machilus rubescens and an application of the Nerolidol / Linalool synthetase gene MnTPS6. The machilus nerolidol / Linalool synthetase gene MnTPS6 is a key gene for synthesis of the nerolidol / Linalool in plant genetic engineering, synthesis of the nerolidol / Linalool can be effectively regulated and controlled, expression protein of the machilus nerolidol / Linalool synthetase gene MnTPS6 is applied to synthesis and preparation of the nerolidol / Linalool, the yield is high, and the machilus nerolidol / Linalool synthetase gene MnTPS6 has important application value. Substrates of farnesyl pyrophosphate (FPP) and geranyl pyrophosphate (GPP) are added to the expression protein of the induced machilus nigrum Nerolidol / Linalool synthetase gene MnTPS6, the catalytic activity of the recombinant protein is detected, and a GC-MS (Gas Chromatography-Mass Spectrometer) detection component shows that the result shows that the MnTPS6 is a bifunctional enzyme for synthesizing Nerolidol / Linalool.

The present invention relates to a protein, comprising the following 1)-3): 1) an amino acid sequence as shown in SEQ ID NO.1; 2) a sequence having the same function as the sequence as shown in SEQ ID NO.1 and obtained by replacing, deleting or adding one or more amino acids; 3) a fusion protein having the same function as the protein as shown in SEQ ID NO.1 and connected to a tag at the N-terminus or C-terminus; and also discloses the use of the AarTPS89 gene in catalyzing the precursor substance geranyl pyrophosphate GPP to generate borneol diphosphate, and further generating borneol through the catalytic action of phosphatase; wherein the AarTPS89 gene sequence is as shown in SEQ ID NO.2.

The invention belongs to but is not limited to the technical field of plant genetic engineering, and discloses a beta-Copaene / Geraniol synthetase gene MnTPS11 of Machilus machilus and an application of the beta-Copaene / Geraniol synthetase gene MnTPS11. The beta-Copaene / Geraniol synthase gene MnTPS11 of Machilus machilus provided by the invention is a key gene for synthesis of beta-Copaene / Geraniol in plant genetic engineering, synthesis of beta-Copaene / Geraniol can be effectively regulated and controlled, expression protein of the beta-Copaene / Geraniol synthase gene MnTPS11 is applied to synthesis and preparation of beta-Copaene / Geraniol, the yield is high, and the beta-Copaene / Geraniol synthase gene MnTPS11 has important application value. Substrates of farnesyl pyrophosphoric acid (FPP) and geranyl pyrophosphoric acid (GPP) are added to the expression protein of the induced Machilus machilus beta-Copaene / Geraniol synthetase gene MnTPS11, the catalytic activity of the recombinant protein is detected, and a GC-MS (Gas Chromatography-Mass Spectrometer) detection component shows that the result shows that the MnTPS11 is a bifunctional enzyme for synthesizing beta-Copaene / Geraniol.

The present application relates to the field of synthetic biology technology, and specifically discloses a cannabinol-producing microbial cell, its construction method, and application. A cannabinol-producing microbial cell includes a host cell, and the host cell includes a synthetic pathway for geranyl pyrophosphate, a synthetic pathway for olivate, and an isopentenyl transferase mutant. The construction method is as follows: based on the geranyl pyrophosphate-producing host cell yCAN10, the olivate synthesis pathway is constructed to form yCG02; a copy gene of the olivate synthase OLS is integrated into yCG02 to obtain yCG03; an acetyl-CoA carboxylase mutant ACC1 (S659A, S1157A) is integrated into yCG03 to obtain yCG04; and an isopentenyl transferase mutant is integrated into yCG04 to obtain yCG05, i.e., a cannabinol-producing microbial cell. The application is to produce cannabinol from a carbon source, which alleviates the problem of a large gap in cannabinol resources in the current market.

The application relates to an application of a geranylgeranyl pyrophosphate reductase in preparation of tocopherol, wherein a nucleotide sequence coding the geranylgeranyl pyrophosphate reductase is shown as SEQ ID No. 11, and the application comprises the following steps: under the assistance of coenzyme iron-oxidation-reduction protein and iron-oxygen-reduction protein-NADP+ reductase, the geranylgeranyl pyrophosphate reductase catalyzes the reduction of delta-tocotrienol to generate delta-tocopherol. The geranylgeranyl pyrophosphate reductase has the function of reducing the unsaturated isoprene side chain of tocotrienol, can catalyze the reduction of tocotrienol to generate tocopherol, does not need to depend on the geranyl pyrophosphate precursor in the existing tocopherol biosynthesis pathway, significantly improves the tocopherol synthesis efficiency, and provides a new strategy for the biosynthesis of tocopherol and other tocopherols.

The invention discloses an application of geranyl-geranyl pyrophosphate synthetase in simple renal cyst, provides a development relationship between the geranyl-geranyl pyrophosphate synthetase and the simple renal cyst, and provides a new direction for targeted therapy of SRC by inducing formation of the renal cyst due to deletion of the geranyl-geranyl pyrophosphate synthetase. The invention further discloses that the variation frequency of the rs3806394 located in the GGPS1 promoter is increased in SRC patients, and the research on the pathological mechanism of SRC occurrence is facilitated.

The invention discloses a tobacco terpenoid synthase gene NtTPS2184 and application thereof, and belongs to the technical field of plant molecular biology, the nucleotide sequence of the tobacco terpenoid synthase gene NtTPS2184 is shown as SEQ ID NO.1, the invention provides a recombinant expression vector, the recombinant expression vector is obtained by positively inserting the tobacco terpenoid synthase gene NtTPS2184 between BamHI and SacI sites of pGEX4.0 plasmid, and the recombinant expression vector is used for expressing the tobacco terpenoid synthase gene NtTPS2184. The invention also provides an application of the tobacco terpene synthase gene NtTPS2184 or the recombinant expression vector in the synthesis of linalool and trans-nerolidone by a catalytic substrate, and the terpene synthase gene NtTPS2184 provided by the invention can be massively expressed by using escherichia coli genetically engineered bacteria, so that the yield of the tobacco terpene synthase gene NtTPS2184 is increased, and the yield of the tobacco terpene synthase gene NtTPS2184 is increased. In the presence of a substrate geranyl pyrophosphate, the monoterpene synthase exerts the activity of the monoterpene synthase to catalytically synthesize linalool; and in the presence of a substrate farnesyl pyrophosphate, the activity of the sesquiterpene synthase is exerted, and the trans-nerolidinol is catalytically synthesized.

The present invention relates to the field of bioengineering technology, and particularly relates to an engineered Saccharomyces cerevisiae strain, a method for constructing the same, and applications thereof. The Saccharomyces cerevisiae strain with the preservation number of CGMCC No. 26221 provided by the present invention is obtained by screening on the basis of the chassis strain Saccharomyces cerevisiae SyBE_Sc14D18 after site-directed mutagenesis, atmospheric and room temperature plasma mutagenesis, and adaptive laboratory evolution induced by hydrogen peroxide. On the basis of this strain, the expression of geranyl pyrophosphate synthase TmCrtE is increased and the expression of the Ura3 gene is restored to obtain an engineered Saccharomyces cerevisiae strain with high lycopene yield.

Disclosed are techniques for monoterpenes biosynthesis in engineered microorganisms having a mevalonate pathway, where the microorganisms express an engineered enzyme. The engineered enzyme may include one or more peptides that are selected to alter the active site pocket volume of the engineered enzyme as compared to a wild-type farnesyl pyrophosphate synthase. The engineered enzyme may include a first peptide (such as a linker peptide) fused to a C-terminus of a farnesyl pyrophosphate synthase. Optionally, a second peptide (such as a Myc-tag or a HA-tag) be fused to the first peptide. The method may include exposing the engineered microorganism to a carbon source (such a glucose). The method may include allowing the engineered microorganism to generate (i) a first product from the mevalonate pathway and (ii) the engineered enzyme. The method may include forming a monoterpene by allowing the first product and the engineered enzyme to interact.

The invention belongs to the technical field of plant genetic engineering, and discloses a machilus nanmu TPS synthetase gene MnTPS7 and an application of the machilus nanmu TPS synthetase gene MnTPS7. The Machilus davidii Sesquiabinene / Linalool synthetase gene MnTPS7 provided by the invention is a key gene for synthesis of Sesquiabinene / Linalool in plant genetic engineering, synthesis of Sesquiabinene / Linalool can be effectively regulated and controlled, expression protein of the Machilus davidii Sesquiabinene / Linalool synthetase gene MnTPS7 is applied to synthesis and preparation of Sesquiabinene / Linalool, the yield is high, and the Machilus davidii Sesquiabinene / Linalool synthetase gene MnTPS7 has important application value. According to the present invention, substrates such as farnesyl pyrophosphate (FPP) and geranyl pyrophosphate (GPP) are added to the expression protein of the induced machilus machilus Sesquiabinene / Linalool synthase gene MnTPS7, the catalytic activity detection of the recombinant protein is performed, the gas chromatography-mass spectrometer (GC-MS) detection component shows that the main catalytic product of the FPP is Sesquiabinene, the main catalytic product of the GPP is Linalool, and the MnTPS7 is the bifunctional enzyme for synthesizing the Sesquiabinene / Linalool;

Provided herein are compositions and methods for producing myrcene by culturing genetically modified microbial host cells that express a myrcene synthase and optionally a geranyl pyroplosphate synthase. Also provided herein are isolated nucleic acid molecules that encode myrcene synthase variants derived from the Ocimum species myrcene synthase, which comprise one or more amino acid substitutions that improve in vivo performance of myrcene synthase in genetically modified microbial host cells. Also provided herein are isolated myrcene synthase variants that exhibit an improved activity for converting geranyl diphosphate into myrcene.

The invention discloses a saccharomyces cerevisiae engineering bacterium for efficiently synthesizing mogroside V based on cytoplasm and peroxisome double-cell-region indoor and a construction method of the saccharomyces cerevisiae engineering bacterium. Wherein the construction method comprises the following steps: integrating acetoacetyl coenzyme A thiolase ERG10, 3-hydroxy-3-methylglutaryl coenzyme A synthetase ERG13, truncated 3-hydroxy-3-methylglutaryl coenzyme A reductase tHMG1, phosphomevalonate kinase ERG8, mevalonate pyrophosphate decarboxylase ERG19, pentenyl pyrophosphate isomerase IDI1 and geranyl pyrophosphate synthase ERG20 at a site of a saccharomyces cerevisiae genome GAL80, and carrying out enzymolysis, so as to obtain a saccharomyces cerevisiae strain; a squalene synthase ERG9, an epoxy squalene cyclization enzyme ERG1 and the like; aBC efflux protein PDR11, isopentene phosphokinase AtIPK, geranyl pyrophosphate synthase ERG20, endogenous squalene synthetase ERG9, epoxy squalene cyclization enzyme ERG1 and the like are integrated at a site Exg1 of a saccharomyces cerevisiae genome. The yield of MG-V synthesized by shake flask fermentation of the saccharomyces cerevisiae engineering bacteria constructed by the invention reaches 117.6 mg / L, and the saccharomyces cerevisiae engineering bacteria have the advantages of high yield, strong metabolic flux directionality, wide industrial application prospect and the like.

The invention provides a method for producing a terpene or a precursor thereof by microbial fermentation. Typically, the method involves culturing a recombinant bacterium in the presence of a gaseous substrate whereby the bacterium produces a terpene or a precursor thereof, such as mevalonic acid, isopentenyl pyrophosphate, dimethylallyl pyrophosphate, isoprene, geranyl pyrophosphate, farnesyl pyrophosphate, and / or farnesene. The bacterium may comprise one or more exogenous enzymes, such as enzymes in mevalonate, DXS, or terpene biosynthesis pathways.

Disclosed herein are isoprenyltransferases capable of producing bakuchiol or an analog thereof from geranyl pyrophosphate (GPP) and cinnamic acid, cumaric acid, caffeic acid and / or ferulic acid. Also disclosed herein are engineered cells expressing an isoprenyltransferase capable of producing bakuchiol or an analog thereof, and methods of producing bakuchiol or an analog thereof using the isoprenyltransferase and the cells.

A prenyltransferase capable of producing bakuchiol or its analogues from geranyl pyrophosphate (GPP) and cinnamic acid, coumaric acid, caffeic acid and / or ferulic acid is disclosed herein. Manipulated cells expressing a prenyltransferase capable of producing bakuchiol or its analogues, as well as methods of using the prenyltransferase and cells to produce bakuchiol or its analogues, are also disclosed herein.

This disclosure generally relates to the use of enzyme combinations or recombinant microbes comprising same to make isoprenoid precursors, isoprenoids and derivatives thereof including prenylated aromatic compounds. Novel metabolic pathways exploiting Claisen, aldol, and acyloin condensations are used instead of the natural mevalonate (MVA) pathway or 1-deoxy-d-xylulose 5-phosphate (DXP) pathways for generating isoprenoid precursors such as isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), and geranyl pyrophosphate (GPP). These pathways have the potential for better carbon and or energy efficiency than native pathways. Both decarboxylative and non-carboxylative condensations are utilized, enabling product synthesis from a number of different starting compounds. These condensation reactions serve as a platform for the synthesis of isoprenoid precursors when utilized in combination with a variety of metabolic pathways and enzymes for carbon rearrangement and the addition / removal of functional groups. Isoprenoid alcohols are key intermediary products for the production of isoprenoid precursors in these novel synthetic metabolic pathways. These precursors can be modified to various isoprenoid products through prenyl transferase, terpene synthase, or terpene cyclases. The production of prenylated aromatic compounds is achieved through prenyl transfer of the hydrocarbon units of isoprenoid precursors to polyketides.

The invention discloses thiogeranyl geranyl pyrophosphate GGSPP and application thereof in preparation of liver cancer drugs, the structural formula of the GGSPP is as shown in formula (I): # imgabs0 # GGSPP specifically binds and activates human fructose-1, 6-diphosphatase 1FBP1 protein, up-regulates the enzymatic activity of FBP1, and reverses the Warburg effect of tumor cells, thereby inhibiting proliferation, migration and development of hepatocellular carcinoma. The invention not only clarifies the key effect of the GGSP-FBP1 axis in the occurrence and development of liver cancer, but also provides an efficient, specific and mechanism-clear anti-liver cancer candidate drug, opens up a new direction for the treatment of metabolism-related tumors, and has important scientific significance and clinical application prospects.

The invention relates to the technical field of metabolic engineering, in particular to a recombinant saccharomyces cerevisiae strain for producing cannabinoid phenolic acid at high yield by using sugar as well as a construction method and application of the recombinant saccharomyces cerevisiae strain. According to the invention, firstly, reverse beta-oxidation pathway genes ReBktB, CnPaaH1, CaCrt and TdTer are introduced, so that the problem that cannabinoid phenolic acid biosynthesis depends on a hexanoic acid substrate is solved; and then the gene GAL80 of the GAL4 inhibitory protein is knocked out, and the GAL4 is over-expressed, so that the overall expression level of the pathway is up-regulated. Then, a mercaptase gene Ckth1A and a hydroxyacyl coenzyme A dehydrogenase gene Ckhbd1 are introduced, so that the supply of the hexanoyl coenzyme A is improved. Furthermore, the endogenous farnesyl pyrophosphate synthetase gene ERG20 is down-regulated, and the consumption of geranyl pyrophosphate by a branch path is reduced. Finally, the expression level of part of genes in saccharomyces cerevisiae cells is regulated and controlled through gene editing, and the purposes of synthesizing cannabinoid phenolic acid from the beginning by using saccharomyces cerevisiae and taking sugar as a substrate and improving the yield of cannabinoid phenolic acid are achieved.

Provided are various enzymes for cannabinoid synthesis and methods for their preparation and use. Also provided are recombinant polypeptides having prenyltransferase activity and / or geranyl diphosphate:olivetolate geranyltransferase (GOT) activity, and engineered recombinant microorganisms for the production of CBGA, and methods for producing cannabigerolic acid (CBGA) or an analogue thereof, the methods comprising the step of reacting a heterologously expressed prenyltransferase with geranyl diphosphate (GPP) and an acid such as olivetolic acid (OA).

The invention provides a yarrowia lipolytica engineering strain with high yield of beta-carotene and construction of the yarrowia lipolytica engineering strain, and belongs to the technical field of synthetic biology and metabolic engineering. According to the invention, a multi-copy geranyl-geranyl pyrophosphate synthase gene, a compartmentalized localization expression beta-carotene synthetic pathway related enzyme gene, a diacylglycerol acyltransferase gene, a cell cycle regulation transcription factor, an oxidative stress regulation related gene, and a cell wall and membrane lipid homeostasis regulation related gene are integrated; and regulating foreign protein folding and expression related genes to obtain the yarrowia lipolytica engineering strain for efficiently biosynthesizing beta-carotene by taking glucose as a substrate. The yield of beta-carotene of the constructed yarrowia lipolytica engineering strain is high, the yield at the shake flask level can reach 3253.2 mg / L, and the yield can be increased to 26.5 g / L when fed-batch fermentation is carried out on a 5L scale. The method has great practical significance and economic value for industrial and commercial application of high-value terpenoid products.

The application discloses a brewing yeast engineering strain for producing bakuchiol as well as a construction method and application thereof. The engineering strain overexpresses a gene in a geranyl pyrophosphate synthesis pathway, a gene in a p-coumaric acid synthesis pathway and a mutant isopentenyl transferase PcPT07 coding gene; the engineering strain has the ability to synthesize bakuchiol from glucose as a single carbon source; the classification and naming of the engineering strain is Saccharomyces cerevisiae BAK07, and the preservation number is CGMCC NO.31081. The application realizes the de novo synthesis of bakuchiol in the brewing yeast through overexpression of the gene in the geranyl pyrophosphate synthesis pathway, the gene in the p-coumaric acid synthesis pathway and the mutant isopentenyl transferase PcPT07 and combining metabolic engineering and other strategies, and lays a foundation for producing bakuchiol through microbial fermentation.

The invention discloses application of a masson pine geranyl and geranyl pyrophosphate synthetase gene PmGGPPS14, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the geranyl and geranyl pyrophosphate synthetase gene PmGGPPS14 of the masson pine is as shown in SEQ ID NO. 1. The amino acid sequence of the expression protein of the PmGGPPS14 is as shown in SEQ ID NO. 2. The invention finds that PmGGPPS14 can promote accumulation of photosynthetic pigments so as to promote plant development and increase leaf area. Overexpression of PmGGPPS14 can improve expression of partial genes in a GGPP downstream pathway, and plays an important role in synthesis of diterpenoid resin acid. The PmGGPPS14 gene provided by the invention is helpful for deep research on the functions of the PmGGPPS14 gene of the pinus massoniana, and provides a molecular means and basis for breeding work for improving synthesis of terpenoid compounds of the pinus massoniana.

Linalool synthase variants are disclosed. The variants preferably comprise an amino acid sequence having one or more amino acid substitutions. Variants producing a greater linalool yield and / or higher linalool:nerolidol ratio compared with the corresponding wild type linalool synthase are also disclosed. A method comprising the conversion of geranyl pyrophosphate to linalool using a linalool synthase variant is also disclosed.

This invention relates to a genetically engineered strain for synthesizing retinal, its construction method, and its applications. The construction method involves introducing DNA fragments of the β-carotene 15,15'-oxygenase gene, isopentenyl pyrophosphate isomerase gene, phytoene dehydrogenase gene, geranyl-geranyl pyrophosphate synthase gene, and phytoene synthase gene into *Rhizopus cylindrica* to obtain a genetically engineered strain. This strain enhances the expression of β-carotene 15,15'-oxygenase, isopentenyl pyrophosphate isomerase, phytoene dehydrogenase, geranyl-geranyl pyrophosphate synthase, and phytoene synthase in *Rhizopus cylindrica*. By selecting *Rhizopus cylindrica* as a host, introducing key synthetic genes, and achieving stable genome integration, this method yields an engineered strain capable of efficiently synthesizing retinal using glucose as a carbon source, simplifying the production process, reducing costs, and improving industrial applicability.

Various aspects of this disclosure relate to compositions comprising (i) one or more decarboxylated cannabinoids and (ii) either (a) cellulose I, (b) nucleic acids that comprise one or more nucleotide sequences that encode a geranyl-pyrophosphate-olivetolic acid geranyltransferase, (c) protein that comprises one or more amino acid sequences that encode a geranyl-pyrophosphate-olivetolic acid geranyltransferase, (d) two of a, b, and c, or (e) each of a, b, and c.

This application relates to a recombinant Saccharomyces cerevisiae for producing cannabigerolic acid, which is prepared from a basic Saccharomyces cerevisiae, wherein the following endogenous genes of the basic Saccharomyces cerevisiae are overexpressed, downregulated or knocked out: pep4 , gal80 , gal4 and fas1 ; exogenous genes encoding the following enzymes are overexpressed: tetraketo synthase ( Cs. TKS) gene, olivetolic acid cyclase ( Cs. OAC) gene, geranyl diphosphate:olivetolate geranyltransferase ( Cs. PT4n) gene and variant acetyl-CoA synthase ( Se. ACS) gene. This application uses Saccharomyces cerevisiae capable of synthesizing CBGA as the original strain, selectively overexpresses Cs. TKS, Cs. OAC, Cs. PT4, Se. ACS, selectively knocks out gal80 and overexpresses gal4 , selectively knocks out pep4 , selectively overexpresses OAC with or without linker fusion of Cs. , localizes Cs. PT4n enzyme to the plasma membrane with or without signal peptide, balances the metabolic flux, optimizes the heterologous synthesis pathway, and significantly improves the yield of CBGA.