75 results about "Peroxisome" patented technology

The invention belongs to the technical application field of microbial metabolic engineering and synthetic biology, and particularly relates to a pichia pastoris engineering strain for high-yield sclareol as well as a construction method and application of the pichia pastoris engineering strain. The method comprises the following steps: constructing a biosynthetic pathway of sclareol in a host strain, and optimizing an intracellular mevalonic acid metabolic pathway and a central metabolic pathway to obtain an engineering strain A; the host strain is pichia pastoris; or overexpressing or knocking out a metabolism regulation factor in the obtained engineering bacterium A to obtain an engineering bacterium strain B; or, in the obtained engineering bacterium B, a synthetic pathway is targeted to peroxisome and optimized by adopting a cell region compartment strategy, and an engineering bacterium strain C is obtained. The invention provides a pichia pastoris chassis cell for synthesizing a diterpenoid sclareol and an engineering strain for synthesizing sclareol. By introducing and optimizing the expression of a sclareol synthesis pathway, the fermentation yields of sclareol in shake flask batch and bioreactor batch fed-batch fermentation are respectively 631.6 mg / L and 10.5 g / L.

The invention provides a high-yield GPP / FPP saccharomyces cerevisiae engineering strain as well as a construction method and application thereof, and relates to the technical field of synthetic biology. Through a gene editing technology, heterologous mvaE and mvaS genes are introduced into saccharomyces cerevisiae, endogenous ERG12, ERG8, ERG19 and ERG20 genes are over-expressed, and the metabolic flux of an MVA pathway in saccharomyces cerevisiae cells is enhanced; meanwhile, a positioning signal tag of peroxisome is added to the gene, and an MVA pathway and a terpenoid precursor synthesis pathway are reconstructed in the peroxisome, so that the compartment synthesis of the terpenoid precursor is realized. Besides, a heterologous MthIPKs gene is further introduced, a branch pathway for IPP synthesis is constructed, higher energy efficiency and less enzymatic reactions are achieved, an MVA pathway and metabolic flux for terpenoid synthesis are concentrated, and the biosynthesis efficiency and conversion rate are further improved.

The invention provides an application of a pentacyclic triterpenoid compound targeting hydroxyl steroid 17-beta dehydrogenase 4. The pentacyclic triterpenoid compound can effectively relieve or treat central nervous system diseases (inflammatory diseases), including central nervous system degenerative diseases or chronic central inflammatory diseases. The pentacyclic triterpenoid compound can maintain the structural stability or activity of hydroxysteroid 17-beta dehydrogenase and maintain the stability or activity of peroxisome, thereby relieving or treating central nervous inflammation. The pentacyclic triterpenoid compound can target hydroxysteroid 17-beta dehydrogenase 4, inhibit neuroglial cell excessive activation related neuroinflammation, relieve neuron damage or death speed and improve a central nervous system cell metabolism microenvironment.

The invention discloses candida tropicalis engineering bacteria for high yield of (-)-alpha-bisabolol and a construction method of the candida tropicalis engineering bacteria, and belongs to the technical field of microorganisms. According to the invention, chassis cell candida tropicalis DC04-WG04-2 is constructed, a gene Cat2 of catalase derived from saccharomyces cerevisiae and a gene ERG10 of 3-hydroxy-3-methylglutaryl coenzyme A reductase are expressed on the basis of the chassis cell candida tropicalis DC04-WG04-2, and a PEX15 motif is anchored on the surface of peroxisome for co-expression; the genes ADH1, GPD1 and GPD2 are further knocked out, so that the accumulation of ethanol is reduced, and the generation of glycerol is reduced. According to the method disclosed by the invention, the yield of the (-)-alpha-bisabolol of the recombinant candida tropicalis reaches 461.7 mg / L by optimizing fermentation conditions. The preparation method has the characteristics of low cost, high efficiency, simplicity in operation, wide prospect and the like.

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.

Provided are a conjugate of the protein ATF6αN and peroxisomes, a use of the protein ATF6αN in the isolation and purification of peroxisomes, and a method for isolating and purifying peroxisomes by using the protein ATF6αN. The method comprises the following steps: combining a tag-ATF6αN fusion protein with a medium, so as to obtain a fusion protein-medium complex; adding a crude peroxisome sample to the fusion protein-medium complex, so as to obtain a fusion protein-medium-peroxisome complex; and adding an elution buffer and collecting an eluent, so as to obtain peroxisomes conjugated with a fusion protein.

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 discloses a hydrogel composite material for repairing acute kidney injury as well as preparation and application thereof. The hydrogel composite material for repairing the acute kidney injury comprises peroxisome serving as an oxidative stress regulation center and self-assembly repairing polypeptide gel for encapsulating the peroxisome. According to the composite material, oxidative stress regulation is carried out through a peroxisome cascade catalytic system, a polypeptide precursor solution is initiated to be self-assembled and polymerized into gel through pH responsiveness, local drug delivery is carried out, and cell repair regulation and control of kidney injury are achieved. The problem that oxidative stress and tissue damage are difficult to solve at the same time in existing single drug treatment is solved, wound healing is accelerated by rapidly neutralizing free radicals, reducing tissue damage, activating cell proliferation and regenerating related signal channels, the composition can act on damaged tissue more accurately, the influence on healthy cells is reduced, and due to the synergistic effect mechanism, the composition has a good application prospect. More efficient and targeted kidney repair can be realized, so that remarkable advantages are shown in the repair treatment of the acute kidney injury.

The invention belongs to the technical field of bioengineering, and particularly relates to an engineering bacterium for de novo biosynthesis of cycloastragenol as well as a construction method and application of the engineering bacterium. A mevalonic acid (MVA) pathway is reconstructed in a saccharomyces cerevisiae strain peroxidase body by utilizing a cell region compartment strategy, a key enzyme gene of the MVA pathway is over-expressed by using a strong promoter, and the cycloastragenol is obtained. The key enzyme genes are used as expression elements of cycloastragenol, are introduced into the chassis saccharomyces cerevisiae through a CRISPR-Cas9 technology, and are integrated on a saccharomyces cerevisiae genome through homologous recombination, so that a saccharomyces cerevisiae engineering bacterium CycZ23 and an engineering bacterium CAG-1 for efficiently synthesizing cycloastragenol are constructed, and de novo synthesis of plant triterpenoid natural products is realized; the highest titer of the generated cycloastragenol is 236 mg / L; the synthesis method is economical, efficient, green, environmentally friendly and easy to operate, and has important application value.

The invention relates to the technical field of biology, in particular to engineered yeast for high-yield citronellol as well as a preparation method and application of the engineered yeast. According to the invention, genetically engineered bacteria for high yield of citronellol are constructed by means of genetic engineering, SyBESc081300004 is taken as an original strain, Erg20ww, Erg8, tCrGES and CrIS are expressed in a peroxisome region in a room manner, and phosphopentose pathway genes TAL1 and TKL1 and endogenous transporter protein PDR1 are over-expressed, so that the genetically engineered bacteria are obtained; experimental results show that the shake flask fermentation yield of the genetically engineered bacterium reaches 3.38 g / L, the yield of a 100L fermentation tank reaches 10.556 g / L, and the yield of the genetically engineered bacterium is obviously higher than that of other existing citronellol production engineered bacterium.

The invention relates to a method for reducing body weight of a subject comprising exposing a skin area of the subject to a sub-erythemal dose of narrow-band UV-B radiation in the range of 305-315 nm. The invention further relates to a narrow-band UV-B lamp for use in the treatment and / or prevention of obesity and / or metabolic syndrome in a subject, wherein the lamp emits UV-B radiation in the range of 305-315 nm and is preferably configured for whole body irradiation of a human subject. In another aspect the invention relates to UV-B radiation for use in the treatment and / or prevention of obesity and / or metabolic syndrome in a subject, wherein the UV-B radiation is in the range of 305-315 nm. In embodiments, the narrow-band UV-B radiation induces transient / reversible metabolic remodeling involving one or more of mitochondrial fragmentation / fission, peroxisomal and mitochondrial lipid beta-oxidation, lipid droplet turnover and glycolysis.

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 contents disclosed herein include methods, compositions, and kits suitable for use in enhanced adoptive T cell therapy. In some embodiments, the methods include contacting an isolated organelle complex with a population of T cells to produce a population of T cells comprising the organelle complex. The organelle complex can include mitochondria and one or more of endoplasmic reticulum, peroxisome, lysosome, and Golgi. The population of T cells exhibits one or more of the following: enhanced expansion capability, enhanced cytotoxicity against a target cell, enhanced resistance to depletion, and enhanced persistence compared to a population of T cells not comprising an exogenous organelle complex.

The invention discloses a saccharomyces cerevisiae strain with high yield of campesterol as well as a construction method and application of the saccharomyces cerevisiae strain. One of the key rate-limiting enzymes, which are heterologously synthesized in the yeast, of the campesterol ((24R)-Ergosta-5-en-3beta-ol) is from a plant C-24 (28) reductase DWF1 mutant (Ar207). In view of the characteristics that the catalytic activity of the enzyme is relatively low and substrates are dispersedly distributed in different compartments of cells, on the basis of the original recombinant saccharomyces cerevisiae for producing campesterol, a rate-limiting enzyme gene copy positioned to a specific subcellular organelle is additionally added, so that the campesterol can be produced. The expression quantity is effectively increased; meanwhile, the contact between the enzyme and a substrate in a corresponding chamber is enhanced; and the yield of campesterol is increased. By utilizing the strategy, Ar207 is positioned to recombinant bacteria of peroxisome and lipid droplets, and the yield of campesterol reaches 3.00 g / l and 3.77 g / l respectively during fermentation in a 5L tank.

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.

Provided herein are methods and compositions comprising a bacterium or a metabolite thereof for enhancing mitochondrial and / or peroxisomal function.

The contents disclosed herein include populations of organelle complexes. The organelle complex can include mitochondria and one or more of endoplasmic reticulum, peroxisome, lysosome, and Golgi. In some embodiments, the organelle complexes are isolated from or derived from floating cells and / or frozen cells. In some embodiments, the organelle complexes are isolated from or derived from cells contacted with a surfactant at a concentration equal to or higher than the critical micelle concentration (CMC) of the surfactant. At least about 80% of the mitochondria of the organelle complex is capable of maintaining structural integrity in an extracellular environment. Also provided herein are methods for generating a first population of organelle complexes.

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 technical field of microorganisms, and particularly discloses recombinant saccharomycetes and application thereof in fermentation production of beta-elemene. Compared with a starting yeast strain, the recombinant yeast provided by the invention expresses farnesyl diphosphate synthase ERG20, sesquiterpene synthase LsLTC2 from lettuce and difunctional farnesyl diphosphate synthase RrFPS from rose, and expresses IDI1, ERG13, ERG8, ERG10, ERG12, MVD1 and tHMGR in peroxisome. Preferably, the expression quantity of squalene synthase ERG9 in a cell matrix is further reduced, ERG10, ERG13 and IDI1 are overexpressed, and XFPK, PTA, POXB and ACS are expressed. The recombinant saccharomycetes disclosed by the invention can efficiently produce the beta-elemene, and a more effective method is provided for industrial production of the beta-elemene.

The present invention discloses a genetically engineered Schizochytrium strain for efficient DHA production, as well as a construction method and application. The genetically engineered strain is obtained by knocking out the peroxisome matrix protein gene PEX10 in the wild-type Schizochytrium, which disrupts the β-oxidation pathway and blocks the degradation of fatty acyl-CoA; overexpressing the acetyl-CoA carboxylase gene ACC1 to provide more malonyl-CoA precursors, thereby promoting fatty acid biosynthesis; and overexpressing the diacylglycerol acyltransferase gene DGAT to promote the accumulation of fatty acids in the form of triglycerides. Under shake flask conditions, after fermentation, the DHA and PUFA produced reached 55.10% and 70.47% of the total oil content, respectively. Compared with the wild-type strain, DHA and PUFA increased by 21.6% and 24.77%, respectively, without a significant change in biomass, showing extremely high application potential.