26 results about "Peroxisome" patented technology

The invention discloses a Yarrowia lipolytica engineering bacterium for producing astaxanthin in a peroxisome compartment and application of the Yarrowia lipolytica engineering bacterium. A complete astaxanthin synthesis path is expressed in host bacteria by recombinant Yarrowia lipolytica; comprising geranyl geranyl diphosphate synthase CrtE, phytoene synthetase / lycopene cyclase CrtYB, phytoene desaturase CrtI, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR, beta-carotene ketolase CrtW and beta-carotene hydroxylase CrtZ, and the enzymes are relocated into peroxisome, so that the enzyme activity of the peroxisome is improved, and the enzyme activity of the peroxisome is improved. The number of expression hemoglobin genes VHb and peroxisome is increased, the surface area of the hemoglobin genes VHb is enlarged, and one or more of enzymes PEX11, PEX19, DNM1 or PHO85 for prolonging the service life of the hemoglobin genes VHb and peroxisome is / are obtained; the astaxanthin production performance of the recombinant strain is verified based on cytoplasm engineering and subcellular organelle compartment engineering, and the astaxanthin production capacity of the yarrowia lipolytica is further improved.

This invention discloses a lipophilic yeast strain for synthesizing astaxanthin (Yersinia lipophila). Yarrowia lipolytica This invention relates to an engineered strain, its construction method, and its application. The engineered strain is a *Yersinia lipolytica* strain carrying CrtZ and CrtW. The invention utilizes genetic engineering methods to introduce a *Yersinia lipolytica* strain derived from *Rhodochophora* into a β-carotene-producing strain via genetic engineering. Haematococcus Pluvialis The β-carotene hydroxylase encoding gene (HpCrtZ) originates from the genus Paracoccus ( Paracoccus sp. The gene encoding β-carotene ketolase (PsCrtW) and the gene derived from Saccharomyces cerevisiae (Saccharomyces cerevisiae) Saccharomyces cerevisiae The method involves obtaining a genetically engineered *Yersinia lipolytica* strain that produces astaxanthin by constructing a fusion enzyme complex and fusing it with peroxisomes, endoplasmic reticulum, and lipid droplet-targeting signal peptides to obtain a *Yersinia lipolytica* strain with higher astaxanthin yield. The astaxanthin-producing genetically engineered strain constructed in this invention achieved an astaxanthin yield of 1.43 g / L in a 5 L bioreactor after pH control and continuous feeding optimization.

The invention provides a genetic engineering strain of high-yield immunosuppressant mycophenolic acid as well as a preparation method and application of the genetic engineering strain. The invention firstly provides a method for constructing the penicillium with the effect of high yield of mycophenolic acid. The method comprises the step of replacing an indigenous promoter of acyl-coenzyme A oxidase PbACOX323 protein participating in beta oxidation in peroxisome of a penicillium strain with a promoter PgpdA of 3-phosphoglycerol dehydrogenase with an amino acid sequence of SEQ ID NO: 1. According to the method disclosed by the invention, the genetically engineered bacterium with high yield of mycophenolic acid is constructed, the yield of mycophenolic acid is positively increased by replacing a promoter of acyl-coenzyme A oxidase PbACOX323, and the method has a good industrial application value.

The present invention relates to decanoic acid and a composition comprising the same for use in the treatment and / or prevention of peroxisome disorders.

This invention belongs to the field of microbial metabolic engineering and biotechnology applications, specifically relating to an engineered bacterium for synthesizing monoterpenoid compounds, its construction method, and its applications. The construction method involves using *Hansenula polymorpha* as a host, regulating the mevalonate pathway in its cytoplasm, and overexpressing monoterpene synthase to obtain the engineered bacterium; or, overexpressing monoterpene synthase and the mevalonate pathway in peroxisomes to obtain the engineered bacterium. The engineered bacterium constructed in this application can efficiently synthesize monoterpenoid compounds such as geraniol, β-myrcene, and dextrorotatory borneol.

The invention relates to the technical field of microorganisms, and discloses recombinant yeast for efficiently synthesizing a GPP derivative compound as well as a construction method and application of the recombinant yeast. The recombinant yeast strain takes a yeast strain as an original strain, and is obtained by introducing a target coding gene into a cell nucleus; the target coding gene comprises a coding gene of a mevalonic acid (MVA) synthetic pathway enzyme, a coding gene of a farnesyl diphosphate synthase mutant and a coding gene of optional geranyl diphosphate derivative synthase. According to the invention, an MVA pathway and a key enzyme are introduced into a cell nucleus, so that competitive consumption of a sterol synthesis pathway in cytoplasm on geranyl diphosphate (GPP) is effectively avoided, and efficient synthesis of the GPP derivative compound is realized. Besides, the recombinant yeast strain disclosed by the invention can be combined with a reported peroxisome platform to construct a double-organelle platform strain, so that the yield of GPP derivative compounds can be further increased.

The invention discloses a recombinant Yarrowia lipolytica engineering strain for producing abscisic acid and application of the recombinant Yarrowia lipolytica engineering strain, and belongs to the technical field of genetic engineering and bioengineering. According to the invention, heterologous synthesis of ABA is realized in the Yarrowia lipolytica, precursor supply is optimized by enhancing carbon flow in a metabolic pathway, the activity of key enzymes BcABA2 and BcABA3 is improved by adopting a semi-rational strategy, NADPH regeneration engineering is implemented to enhance the oxygen uptake ability of a strain, a cytoplasm-peroxisome engineering system is constructed, and the oxygen uptake ability of the strain is enhanced. The ABA synthesis is further enhanced by utilizing the characteristic of high-yield acetyl coenzyme A of peroxisome. According to the invention, fermentation conditions are further optimized, so that the ABA accumulation concentration of the constructed engineering strain in a 5 L bioreactor reaches 2554.36 mg / L, and the highest titer of ABA biosynthesis reported so far is created. The metabolic modification strategy provides important reference for biosynthesis of other terpenoids.

Disclosed herein include organelle complexes populations. The organelle complexes can comprise mitochondria and one or more of endoplasmic reticulum, peroxisomes, lysosomes, and Golgi apparatus. In some embodiments, the organelle complexes are isolated or derived from floating cells and / or frozen cells. In some embodiments, the organelle complexes are isolated or derived from cells contacted with a surfactant at a concentration at or above the critical micellar concentration (CMC) for the surfactant. At least about 80% of the mitochondria of the organelle complexes are capable of maintaining structural integrity in an extracellular environment. Also provided herein are methods for generating first organelle complexes populations.

The invention relates to the fields of biochemical engineering, synthetic biology and biological medicine, in particular to a gemmarene A synthetase mutant, a fusion protein, an engineering strain and application. According to the invention, the GASX-L1H2H3V436LH4 with the highest catalytic activity is successfully obtained by means of point mutation and structural domain substitution; compared with wild type LTC2 and HaGAS1, the enzyme has the advantages that the catalytic activity is obviously improved, the fusion protein HaGAS1GGGSERG20 is constructed through a flexible linker, and the beta-elemene synthesis efficiency is further improved; on the basis, by strengthening a cytoplasm MVA pathway and constructing a peroxisome MVA pathway, intracellular precursor substances and acetyl coenzyme A are fully utilized, and the yield of elemene is further increased; on the basis, genes related to grease metabolism are screened to find overexpressed TGL4, and the shake flask yield of elemene is further increased.

The invention discloses a recombinant bacterium and application thereof in production of triptolic acid C or a precursor friedelin thereof. The recombinant strain expresses a gene TwOSC1T502E for synthesizing a tripterygium wilfordii C precursor friedelin, so that de novo synthesis of friedelin is realized, and the yield of the precursor friedelin is increased through MVA (Mycopoeic Acid) pathway optimization and peroxisome metabolism-form collaborative optimization; the method comprises the following steps: by taking tripterygium wilfordii as a raw material, firstly synthesizing tripterygium wilfordii in saccharomyces cerevisiae, then expressing tripterygium wilfordii C synthesis related genes ThCYP712K1 and TrCYP716C52 and TwCPR3, realizing de novo synthesis of tripterygium wilfordii C in saccharomyces cerevisiae, obtaining the ThCYP712K1 mutant ThCYP712K1I124V with improved activity through enzyme engineering, and remarkably improving the yield of tripterygium wilfordii C through lipid droplet engineering modification and cofactor engineering modification. The recombinant strain provided by the invention can be used for efficiently producing the triptolide C and the precursor friedelin of the triptolide C, and has an important application value.

Disclosed herein include methods, compositions, and kits suitable for use in enhancing adoptive T cell therapy. In some embodiments, the method comprises contacting isolated organelle complexes with a population of T cells to generate a population of T cells comprising the organelle complexes. The organelle complexes can comprise mitochondria and one or more of endoplasmic reticulum, peroxisomes, lysosomes, and Golgi apparatus. The population of T cells can exhibit one or more of enhanced expansion capability, enhanced cytotoxicity against target cells, enhanced resistance to exhaustion, and enhanced persistence, as compared to a population of T cells that do not comprise exogenous organelle complexes

The invention discloses kluyveromyces marxianus genetically engineered bacteria and application thereof, and belongs to the technical field of bioengineering. The invention discloses a kluyveromyces marxianus genetically engineered bacterium. The kluyveromyces marxianus genetically engineered bacterium is prepared by the following steps: firstly, screening to obtain a kluyveromyces marxianus strain with high ergosterol yield; on the basis, inducing and separating a MATa haploid strain; then, through the introduction of heterologous DHCR24 and DWF5 genes, the weakening of a competitive pathway gene ERG6, the improvement of the metabolic flux of a squalene pathway in a sterol synthesis process, the compartment engineering of a peroxisome region of a mevalonic acid pathway, the knockout of an hypoxia inhibition transcription factor ROX1, the improvement of cofactor supply, the optimization of a yeast lipid pathway and other metabolic engineering optimization reconstructions, the expression of the yeast lipid pathway is improved; and an engineering strain with high cholesterol yield is constructed and obtained. Through fed-batch fermentation, efficient and stable synthesis of cholesterol is achieved, the yield reaches 7.64 g / L, and a foundation is laid for biological manufacturing of cholesterol.

Method and compositions for producing malonyl-CoA and / or triacetic acid lactone within peroxisomes using genetically modified yeast cells to metabolize a substrate containing fatty acids, vegetable oil, or alkanes, and genetically modified organisms are provided. Also provided are methods to generate acetyl-CoA, which is retained in the peroxisomes due to genetic modifications that prevent its export therefrom. A modified enzyme with a peroxisomal targeting sequence catalyzes a reaction between acetyl-CoA and oxaloacetate, producing pyruvate and malonyl-CoA. The methods can include deleting carnitine acetyltransferase (CAT2) alleles to prevent acetyl-CoA export from peroxisomes, and using genes encoding methylmalonyl-CoA carboxyltransferase subunits with peroxisomal targeting sequences. Additionally, the method incorporates 2-pyrone synthase genes to further facilitate production processes.

The invention discloses a candida tropicalis engineering bacterium for producing astaxanthin, and belongs to the field of metabolic engineering. Candida tropicalis ANT-06 is used as a starting strain to integrally express a heterologous gene CrtZ and a gene CrtW, the heterologous gene CrtZ and the gene CrtW are subjected to fusion expression with a peroxisome signal peptide ePTS and a lipid droplet signal peptide HD2, and the yield of astaxanthin can be obviously increased through a combined strategy of knocking out a phosphatidic acid phosphatase gene PAH1 and a functional homolog gene FLD1 of human seipin. And after combined fermentation optimization, fermentation culture at 22 DEG C and addition of 10% of dodecane, 2.5% of DMSO and 2.5 mM of betaine, the shake flask fermentation yield of the astaxanthin is increased to 320.1 mg / L, and the amplification fermentation yield of a 5L fermentation tank is increased to 3096.2 mg / L. The preparation method has the characteristics of low cost, high efficiency, simplicity in operation, wide prospect and the like.

This invention discloses a genetically engineered strain of *Kluyveromyces martensii* and its applications, belonging to the field of bioengineering technology. The invention discloses a genetically engineered *Kluyveromyces martensii* strain, firstly obtained through screening a strain that produces high levels of ergosterol; based on this, it induces and isolates... Eye A haploid strain was constructed and then metabolically optimized through the introduction of heterologous DHCR24 and DWF5 genes, weakening of the competitive pathway gene ERG6, increasing the metabolic flux of the post-squalene pathway during sterol synthesis, peroxisome compartmentalization engineering of the mevalonate pathway, knockout of the hypoxia-inhibiting transcription factor ROX1, improved cofactor supply, and optimization of the yeast lipid pathway, resulting in a high-cholesterol-producing engineered strain. Through fed-batch fermentation, efficient and stable cholesterol synthesis was achieved, with a yield of 7.64 g / L, laying the foundation for cholesterol biomanufacturing.

The invention belongs to the technical field of biology, and discloses a method for improving the expression quantity of recombinant human serum albumin aiming at the problems of effect limitation and insufficient universality of the existing method for improving the expression quantity of the recombinant human serum albumin, which comprises the following steps: S1, constructing an rHSA expression strain to obtain a candidate strain A / GS115; s2, carrying out multi-gene combination overexpression recombinant engineering bacterium construction by taking the candidate strain A / GS115 as a host bacterium; and S3, fermenting the constructed recombinant engineering bacteria to prepare the recombinant human serum albumin. Related proteins PMP, VMA21, AAP and VTC in peroxisome of pichia pastoris are subjected to overexpression, methanol metabolism and utilization can be promoted, toxicity of toxic substances generated after methanol oxidation to cells is reduced, and therefore the expression quantity of rHSA is increased.

The invention discloses a genetically engineered bacterium for producing phytoene, lycopene and beta-carotene and application of the genetically engineered bacterium, and belongs to the technical field of genetic engineering and metabolic engineering. According to the invention, endogenous phytoene synthase CrtB and phytoene dehydrogenase gene CrtI in rhodotorula host bacteria are knocked out, expression of 2-acylglycerol O-acyltransferase gene DGAT and fatty acid synthase gene FAS is knocked down, and finally, CrtB, CrtI and LCYB are selectively or jointly introduced into cytoplasm-endoplasmic reticulum-peroxisome to obtain the engineering bacteria. According to the engineering bacterium provided by the invention, glucose is used as a carbon source, the yield of phytoene, the yield of lycopene and the yield of beta-carotene are respectively and maximally 82.9 mg / L, 77.1 mg / L and 74.6 mg / L when the engineering bacterium is fermented in a 250 mL shake flask, and the cell dry basis content is more than or equal to 10%; the HPLC purity is greater than or equal to 95%.

The invention discloses a high-yield sclareol recombinant strain as well as a construction method and application thereof, and belongs to the technical field of microbial metabolism engineering. The invention provides recombinant yarrowia lipolytica SC37 with high yield of sclareol, which is characterized in that a sclareol biosynthesis path is firstly constructed, then acetyl coenzyme A in peroxidase is guided to a mevalonic acid path, and then a precursor acetyl coenzyme A and a cofactor NADPH are introduced by optimizing an intracellular mevalonic acid metabolism path to obtain the recombinant strain SC37. The invention provides a recombinant strain SC37 which takes waste oil as a substrate and is high in sclareol yield, and by introducing and optimizing the expression of a sclareol synthesis path, the sclareol yields in shake flask batch fed-batch fermentation and bioreactor batch fed-batch fermentation respectively reach 2.5 g / L and 20.18 g / L. The recombinant strain SC37 has important values for reducing environmental pollution and increasing waste oil economy, and has a good application prospect.

An oil-in-water microemulsion is provided. The microemulsion contains 1-5% v / v oil, 10-25% v / v of at least one emulsifier, 1-10% v / v of at least one co-emulsifier, and a peroxisome proliferat or- activated receptor-a (PPARa) agonist. Methods of preparing the microemulsion and for the ocular delivery of the microemulsion are also provided. The microemulsion may be used for the treatment of ocular injury or ocular degeneration.

The application discloses a kind of hydrogel composite material for acute kidney injury repair, its preparation and application, and the hydrogel composite material for acute kidney injury repair includes peroxisome as oxidative stress regulation center and self-assembled repair polypeptide gel encapsulating peroxisome.The above-mentioned composite material is carried out oxidative stress regulation and pH-responsive polypeptide precursor liquid self-assembly polymerization into glue by peroxisome cascade catalytic system, and local administration is realized to realize cell repair control to kidney injury.The application solves the problem that existing single drug treatment is difficult to simultaneously solve oxidative stress and tissue damage, by neutralizing free radicals quickly, reducing tissue damage, activating cell proliferation, regeneration-related signal pathway, accelerating wound healing, and can more accurately act on damaged tissue, reduce the influence on healthy cells, due to its synergistic mechanism, can realize more efficient and more targeted kidney repair, thereby showing significant advantages in the repair treatment of acute kidney injury.

The application discloses a linalool synthetase t67OMcLIS obtained through protein rational design M The application discloses a mutant gene and a protein encoded by the mutant gene. M The application also discloses a recombinant vector and a recombinant bacterium containing the mutant gene and the protein, and a construction method and application of the mutant gene and the protein. M The application discloses a recombinant bacterium containing the mutant gene and the protein, and the recombinant bacterium can be used for efficiently producing linalool. The application discloses a Saccharomyces cerevisiae diploid strain, which fully utilizes acetyl coenzyme A as a precursor substance in cytoplasm and organelles, simultaneously introduces a linalool synthesis pathway into cytoplasm and peroxisomes of the Saccharomyces cerevisiae, and coordinately regulates the cytoplasm and the peroxisomes, so that the linalool is more efficiently produced.

Disclosed herein include methods, compositions, and kits suitable for use in reducing or preventing oxidative stress and / or reductive stress. In some embodiments, the method comprises contacting a redox-sensitive composition with an effective amount of a disclosed redox-modulating composition. In some embodiments, the method comprises administering to a subject in need an effective amount of a disclosed redox-modulating composition. In some embodiments, the redox-modulating composition comprises isolated organelle complexes. Organelle complexes can comprise mitochondria and one or more of endoplasmic reticulum, peroxisomes, lysosomes, and Golgi apparatus. In some embodiments, the redox-modulating composition comprises isolated mitochondria. The redox-modulating composition can be capable of reducing levels of ROS and / or RNS in a redox-sensitive composition and / or cells.

This invention relates to the field of microbial technology, specifically disclosing a recombinant yeast strain and its application in the fermentation production of β-elemene. Compared to the original yeast strain, the recombinant yeast strain of this invention expresses farnesyl diphosphate synthase ERG20, lettuce-derived sesquiterpene synthase LsLTC2, and rose-derived bifunctional farnesyl diphosphate synthase RrFPS, and expresses IDI1, ERG13, ERG8, ERG10, ERG12, MVD1, and tHMGR in peroxisomes. Preferably, the expression level of squalene synthase ERG9 in the cell matrix is ​​further downregulated, ERG10, ERG13, and IDI1 are overexpressed, and XFPK, PTA, POXB, and ACS are expressed. The recombinant yeast strain of this invention can efficiently produce β-elemene, providing a more effective method for the industrial production of β-elemene.

The invention discloses a 19 peptide PTS2 transport signal small sample prediction system based on incremental learning, and the system comprises a data collection module which is used for collecting a polypeptide sequence, determining a core sequence of PTS2 transport signal peptide, and dynamically intercepting to obtain a 19 peptide fragment; the feature matrix extraction module is used for mapping each amino acid in the 19 peptide fragment into a physicochemical property vector to generate a feature matrix; the PTS2 transport signal prediction model training and prediction module is used for obtaining a PTS2 prediction model based on the feature matrix and through adaptive combination setting of a loss function and incremental training, and carrying out PTS2 prediction; and the PTS2 transfer signal determination module is used for calculating the information entropy based on the prediction result to judge the existence of PTS2. According to the invention, multi-mode characteristics in first 100 amino acids can be effectively identified, accurate positioning and prediction of PTS2 transport signals in a polypeptide sequence are realized, and the method is suitable for protein function analysis and peroxisome targeted drug development.

The application discloses application of OTU4 genes or proteins coded by the genes in improving the high-temperature stress resistance of plants. The application discloses for the first time the influence of a new plant peroxisome protein coded by the OTU4 gene on the heat resistance of plants, and the mutation of the OTU4 gene causes the root growth defect of plants under high temperature. The application discloses for the first time the deubiquitinating enzyme in the peroxisome, and provides a theoretical basis and material support for the ubiquitination research in the peroxisome. The application provides a new gene resource and theoretical guidance for deepening the peroxisome function research and the research and application of the heat resistance of plants and the like.

This invention belongs to the fields of genetic engineering and fermentation engineering, and discloses a method for constructing and applying a Schizochytrium strain that produces high DHA content. The engineered Schizochytrium strain uses Schizochytrium sp. HX308 as the starting strain. Through metabolic engineering, a complete elongation-desaturation pathway is established in Schizochytrium (overexpression of C16 / 18 elongase, Δ-9 elongase gene, Δ-5 desaturase gene, Δ-17 desaturase gene, Δ-5 elongase gene, and Δ-4 desaturase gene), which works synergistically with the PKS pathway to produce DHA. The peroxisome synthesis factor PEX10 is knocked out to obtain the recombinant engineered strain SCD3. In fed-batch fermentation, the final DHA content of the engineered Schizochytrium strain SCD3 is the highest to date, reaching 70.1% of the total fatty acid content. The oil and DHA yields are also significantly increased, reaching 67.9 g / L and 47.6 g / L, respectively. The DHA yield is 1.7 times that of the wild type. This engineered strain has good application prospects.