23 results about "Carotene synthesis" patented technology

The invention provides recombinant yarrowia lipolytica as well as a construction method and application thereof. According to the method disclosed by the invention, beta-carotene synthesis related genes are transferred into a yarrowia lipolytica starting strain to obtain the recombinant yarrowia lipolytica with the capacity of high yield of beta-carotene; furthermore, related genes are introduced on the basis of the recombinant yarrowia lipolytica with high yield of beta-carotene, so that efficient conversion from beta-carotene to zeaxanthin is successfully realized.

The invention belongs to the technical field of genetic engineering, and discloses a schizochytrium limacinum genetic engineering strain for enhancing supply of acetyl coenzyme A and efficiently producing astaxanthin, a method and application of the schizochytrium limacinum genetic engineering strain. By heterologous expression of a beta-carotene ketolase CrtO gene and a beta-carotene hydroxylase CrtZ gene from haematococcus pluvialis, synthesis of astaxanthin from beta-carotene in schizochytrium limacinum is promoted, so that the content of astaxanthin in schizochytrium limacinum HX-308 is increased. According to the invention, the endogenous AACT gene and heterologous CrtO, CrtZ, PK and PTA genes of schizochytrium limacinum are integrated into an original strain through homologous recombination, and the obtained strain still keeps genetic stability after multiple subculture, has the capability of efficiently synthesizing astaxanthin, effectively improves the yield of astaxanthin, and lays a foundation for large-scale industrial production.

The invention provides a genetically engineered bacterium for producing astaxanthin, and the genetically engineered bacterium takes Yarrowia lipolytica Po1f as an original strain and contains a GGS gene, an HMGR gene, a beta-carotene synthesis gene, a beta-carotene ketolase gene and a beta-carotene hydroxylase gene. The invention further provides the Yarrowia lipolytica MC247, and the preservation number of the Yarrowia lipolytica MC247 is CGMCC NO.29699. According to the Yarrowia lipolytica MC247, the high-yield astaxanthin is obtained, a new technical scheme is provided for efficient and green manufacturing of the astaxanthin, and industrial production of the astaxanthin is facilitated.

The invention relates to a beta-carotene production engineering bacterium as well as a construction method and application thereof. According to the engineering bacterium, Yarrowia lipolytica is taken as an original strain, and a beta-carotene synthetic pathway related gene is integrated in a genome of the Yarrowia lipolytica; wherein the beta-carotene synthetic pathway related genes comprise MvaS, MvaE, ERG12, ERG8, ERG19, IDI, ERG20, GGPPs7, CarRP and CarB, and the beta-carotene synthetic pathway related genes comprise beta-carotene, beta-carotene, beta-carotene, beta-carotene, beta-carotene, beta-carotene, beta-carotene and beta-carotene. The beta-carotene synthetic pathway related gene is subjected to localization expression in peroxisome through the peroxisome localization signal peptide. The engineering strain provided by the invention can obviously improve the yield of beta-carotene.

The invention relates to bacillus subtilis with high yield of beta-carotene as well as a construction method and application of the bacillus subtilis, and belongs to the technical field of fermentation engineering. According to the invention, extracellular proteases AprE, NprE, Bpr, Mpr, Epr and NprB are knocked out from bacillus subtilis, a repressor protein mutant tetR expression cassette and a beta-carotene synthetic pathway related gene expression cassette are constructed, and a tetracycline inducible promoter is used for expressing beta-carotene synthetic pathway related genes crtE, crtI, crtB and crtY, so that the bacillus subtilis for producing beta-carotene is successfully constructed. The recombinant strain G600-crtEBIY is constructed, the induction time and the inducer concentration are optimized in the fermentation process of the constructed recombinant strain G600-crtEBIY, the yield of the optimized beta-carotene in a shake flask fermentation system reaches up to 18.5 mg / L, stable and efficient production of the beta-carotene is achieved, and industrial production and popularization of the beta-carotene are facilitated.

The present invention relates to process for producing β-carotene in a specific solvent.

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 invention discloses a pichia pastoris recombinant strain for producing astaxanthin as well as a construction method and application of the pichia pastoris recombinant strain. According to the method, pichia pastoris serves as an operation platform, astaxanthin serves as a target product, a beta-carotene high-yield strain is obtained by increasing the dosage of beta-carotene synthesis genes, then astaxanthin is synthesized through pichia pastoris by integrating astaxanthin synthetase genes, and the astaxanthin yield is improved through optimization of a carotenoid synthesis module. In addition, the synthesis of the recombinant strain astaxanthin is promoted through multienzyme assembly of key nodes in an upstream path of an astaxanthin synthesis path and weakening of a squalene synthesis path in a branch path. On the basis, recombination strain cofactor synthesis is optimized, astaxanthin synthesis is improved, the astaxanthin yield is further increased, and the astaxanthin high-yield strain is obtained. According to the invention, the yield of astaxanthin reaches 4.75 g / L by using a pichia pastoris combined regulation strategy, which is the highest level in current microbial synthesis.

The application discloses a gene related to osmanthus fragrans carotenoid synthesis OfPHL5 and application thereof OfPHL5 The gene is a MYB transcription factor gene, and an overexpression vector containing the gene OfPHL5 is transformed into osmanthus fragrans petals to perform transient expression. OfPHL5 The results show that the gene expression amount of the transiently transformed osmanthus fragrans petals is significantly increased, and the total carotenoid content in the petals is increased, which indicates that the gene plays an important role in promoting osmanthus fragrans carotenoid synthesis. OfPHL5 The gene can be used for improving the color quality of osmanthus fragrans and excellent variety breeding in osmanthus fragrans genetic engineering.

The invention relates to a novel intermediate which can be used in a method for producing beta-carotene.

The invention belongs to the technical field of beta-carotene synthesis, and discloses a fermentation method of a strain for synthesizing beta-carotene, which comprises the following steps: S1, strain preparation and stability enhancement; s2, seed liquid preparation and proportion pre-optimization; s3, dynamic proportion regulation and control in the fermentation process; s4, material supplementation and process optimization; s5, terminating and harvesting; the step of strain preparation and stability enhancement comprises strain selection and activation and genetic stability monitoring, the step of seed solution preparation and proportion pre-optimization comprises a seed culture medium formula and culture parameters, and the step of fermentation process dynamic proportion regulation and control comprises a fermentation culture medium formula, initial inoculation and conditions and a dynamic proportion regulation and control strategy. According to the method, synthesis interruption caused by proportion imbalance is avoided, the effect is derived from the synergistic effect of stabilization of an internal metabolic pathway of the strain and external regulation instead of simple repeated technical steps, the industrial life of the produced strain is remarkably prolonged, and the quality control cost between batches is reduced.

The present invention relates to a method for producing beta-carotene in a specific solvent.

The invention discloses an OfPHL5 gene related to synthesis of osmanthus carotenoid and application of the OfPHL5 gene, the OfPHL5 gene is an MYB transcription factor gene, an overexpression vector containing the gene OfPHL5 is transformed into osmanthus petals for transient expression, and the result shows that the expression quantity of the OfPHL5 gene in the osmanthus petals subjected to transient transformation is remarkably increased, so that the OfPHL5 gene can be applied to the synthesis of the osmanthus carotenoid. The OfPHL5 gene plays an important role in promoting the synthesis of the carotenoids in the osmanthus fragrans, and can be used for improving the color and quality of the osmanthus fragrans, breeding excellent varieties and the like in the genetic engineering of the osmanthus fragrans.

The present invention relates to a process for producing β-carotene in specific solvents.

The invention belongs to the technical field of microorganisms and fermentation engineering, and particularly relates to yarrowia lipolytica engineering bacteria capable of producing beta-carotene at high yield as well as a construction method and application of the yarrowia lipolytica engineering bacteria. Specifically, Yarrowia lipolytica is modified, beta-carotene synthetic pathway related genes are constructed to a genome integrated plasmid, and an original strain is introduced through multiple rounds of transformation. And a diphenylamine color screening method is combined to finally obtain the high-yield beta-carotene strain. The method has the advantages of high yield, few byproducts, low cost, good stability and the like, the production cost is reduced by optimizing the fermentation process, the cost performance is further improved, a foundation is laid for industrial production, and remarkable economic and social benefits are achieved.

The invention provides a regulation and control mechanism of EjERF003 and EjERF029 transcription factors on synthesis of loquat carotenoids and application of the regulation and control mechanism. The relative expression quantity of carotenoid metabolism related genes and ERF transcription factors of loquat fruits treated by 1-MCP is detected through fluorescent quantitative PCR, and it is found that the expression quantity of EjERF003 and EjERF029 transcription factors in the ERF transcription factors is related to the expression quantity of the carotenoid metabolism related genes; an LUC / REN dual luciferase experiment, a yeast single impurity experiment and a gel migration experiment prove that the EjERF003 and the EjERF029 can reversely regulate and control carotenoid structure genes EjPSY2, EjPDS, EjZISO, EjZEP2, EjZDS2 and EjBCH2, and meanwhile, the EjERF003 and the EjERF029 can positively regulate and control a degradation gene EjCCD1. A yeast two-hybrid experiment and a bimolecular fluorescence complementation experiment prove that the EjERF003 transcription factor and the EjERF029 transcription factor can interact with each other and cooperatively regulate the content of carotenoid. The invention also constructs an EjERF003 and / or EjERF029 overexpression vector or interference vector to successfully regulate and control the content of carotenoid in loquat.

The present invention discloses a method for biosynthesizing dihydro-β-ionone, comprising incubating a transformed first host cell in a suitable culture medium containing β-ionone for a sufficient time to convert β-ionone into dihydro-β-ionone; wherein the transformed first host cell comprises a synthetic or recombinant nucleic acid molecule, the nucleic acid molecule comprising a first nucleotide sequence encoding an olefin reductase AaDBR1 from Artemisia annua and a second nucleotide sequence encoding an NADPH regeneration enzyme. The present invention first constructs three modules, namely, a supply module for terpenoid precursors for biosynthesis of β-ionone, a β-carotene synthesis module, and β-ionone synthesis, and recombinantly optimizes the three modules, constructs an excellent recombinant bacterium for synthesizing β-ionone from scratch, and provides a method for realizing green, sustainable, large-scale synthesis of dihydro-β-ionone and its wide application as a natural product.

This invention discloses a stress-induced method for cultivating high-content β-carotene algae, comprising three sequentially executed stages: a balanced growth stage I, used to obtain highly active algal cells; a synergistic induction stage II, to trigger β-carotene synthesis; and an enhanced accumulation stage III, which applies intermittent strong light pulses and temperature oscillations synchronized with the light-dark cycle to increase product accumulation. Stages II and III are implemented in a closed photobioreactor. This invention, through phased, multi-stress synergy and closed-loop precise control, directionally activates and enhances the β-carotene biosynthetic pathway while maintaining high cell activity and biomass. Ultimately, it achieves a volumetric yield ≥50% higher than the traditional high-salt stress method, with cell dry weight not less than 85% of the control, resolving the contradiction between algal cell growth and product accumulation, and possesses promising prospects for industrial application.

The application discloses a preparation method of 2,4-dibond fifteen carbon phosphonate, which uses methylenediphosphonic acid tetraethyl ester and 2-methyl-4-(2,6,6-trimethyl-3-hydroxycyclohexene-1-yl)-2-butene aldehyde (referred to as fourteen carbon aldehyde) as raw materials, and generates Wittig-Horner reaction, and then is hydrolyzed to obtain 2,4-dibond fifteen carbon phosphonate. In the process that the methylenediphosphonic acid tetraethyl ester and the fourteen carbon aldehyde generate Wittig-Horner reaction, a transposition catalyst is introduced, and then the hydrolysis is carried out to obtain 2,4-dibond fifteen carbon phosphonate with a content of more than 98%, which avoids the 2,4-dibond fifteen carbon phosphonate generated by the Wittig-Horner reaction in the prior art, and improves the product quality due to the influence of by-products in a subsequent vitamin A acetate or beta-carotene synthesis process caused by the residual fifteen carbon aldehyde isomer at other positions, and is suitable for industrial production.

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 invention relates to a novel intermediate which can be used in a method for producing beta-carotene.

The invention provides a genetically engineered bacterium for biosynthesis of retinaldehyde and application of the genetically engineered bacterium, and relates to the technical field of genetic engineering.The genetically engineered bacterium comprises an MEP pathway key enzyme expression module, a beta-carotene synthesis key enzyme expression module and a beta-carotene 15, 15 '-oxygenase expression module, an isopentene precursor IPP / DMAPP is continuously supplied through an MEP pathway, and the retinaldehyde is synthesized through the beta-carotene 15, 15'-oxygenase expression module. The method comprises the following steps of: connecting a carotenoid skeleton synthesis module (crtE / crtB / crtI / crtY) to generate beta-carotene, and introducing beta-carotene 15, 15 '-oxygenase (ddblh) to realize central splitting, so that the stoichiometric target of generating two molecules of retinaldehyde from one molecule of beta-carotene is achieved. The technical problem that in the prior art, due to insufficient precursor supply and overweight metabolic burden, the yield of biosynthetic retinaldehyde is low, and industrial requirements are difficult to meet is solved.

The invention belongs to the technical field of rice gene engineering, and particularly relates to an OsGGPPS oriented mutant protein GGPPS22 / 26. The two sites 22 and 26 of the existing OsGGPPS protein are derived from ancestor sequence reconstruction; on the basis of the two sites, the invention provides an OsGGPPS series oriented mutant protein, which comprises a series of single site mutant protein (OsGGPPS-22 and OsGGPPS-26) and a double site mutant protein (OsGGPPS-22-26). According to the invention, a small and fine mutant library is constructed, specific site amino acid mutation types are analyzed in detail through further screening and based on directed evolution requirements, and a preliminary experimental result shows that after specific site amino acid mutation, the synthesis amount of beta-carotene is obviously increased; and a certain technical foundation is laid for further cultivation of new varieties of crops.