15 results about "Astaxanthin synthesis" patented technology

The invention provides a phaffia rhodozyma mutant strain for high yield of astaxanthin as well as a screening method and application of the phaffia rhodozyma mutant strain, belongs to the technical field of fermentation microorganism screening, and can solve the problems of low astaxanthin yield, high cost and low growth temperature of wild type phaffia rhodozyma existing in synthesis of astaxanthin from a phaffia rhodozyma strain. The invention provides a phaffia rhodozyma mutant strain capable of producing astaxanthin at high yield, the phaffia rhodozyma mutant strain is a phaffia rhodozyma AA0529 mutant strain, the phaffia rhodozyma AA0529 mutant strain is preserved in China Center for Type Culture Collection in Wuhan, Hubei on September 7, 2023, and the preservation number is CCTCC (China Center for Type Culture Collection) NO: M20231637. According to the method, the yield of the astaxanthin can be increased, and the yield of the astaxanthin is increased to 3.3 mg / g at 26 DEG C.

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.

The invention provides a method for promoting synthesis of microcystis astaxanthin by adjusting the concentration of a stress substance, which comprises the following steps of: firstly, culturing microcystis algal bloom or microcystis aeruginosa to a logarithmic phase by adopting a BG11 culture medium under the conditions of specific temperature, illumination and shaking table rotating speed, and adjusting the cell density by centrifuging and resuspending; then, NaCl, KCl, beta-violet ketone, longifolene and H2O2 with different concentrations are added into the algae liquid respectively, control is set, through multi-dimensional index measurement, the algae cell density, the ROS level, the photosynthetic pigment content, the photosynthetic performance and the level of astaxanthin and a precursor thereof are monitored when treatment is carried out for 0 day, 2 days, 4 days and 6 days, the expression quantity of astaxanthin biosynthesis related genes is further analyzed, and the astaxanthin biosynthesis related genes are obtained. Therefore, the optimal concentration of NaCl, KCl, beta-violet ketone, longifolene and H2O2 is determined, the effect of promoting astaxanthin synthesis is compared, and a theoretical basis and a precise regulation and control method are provided for industrial production of astaxanthin by using microcystis.

The invention provides a vector and a method for expressing astaxanthin by using nicotiana benthamiana leaves, which are characterized in that plant virus expression vectors of calendula astaxanthin synthesis related genes AdKC and AdKeto and plant virus gene silencing (RNAi) vectors of nicotiana benthamiana carotenoid cleavage dioxygenase 1 (CCD1) are respectively constructed; the agrobacterium tumefaciens of the three vectors are mixed and infiltrated into the boeng tobacco leaves to obtain the leaves for expressing the astaxanthin. According to the invention, the expression of the nicotiana benthamiana CCD1 gene is interfered while the AdKC and AdKeto genes are expressed in the nicotiana benthamiana leaves, so that the content of astaxanthin expressed in the nicotiana benthamiana leaves is increased.

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 invention discloses application of oxygen-carrying protein VHb in construction of high-yield astaxanthin engineering bacteria, and belongs to the technical field of biology. According to the method, carRPY27R, carB, fusion genes crtW-crtZ and GGPPS are iteratively integrated onto a yarrowia lipolytica chassis genome by utilizing a synthetic biological technology, so that two recombinant strains for producing astaxanthin are obtained. On the basis, the oxygen-carrying protein VHb from vitreoscilla is expressed, and compared with the oxygen-carrying protein VHb which is not expressed, the VHb is positioned to the endoplasmic reticulum, so that the accumulation level of astaxanthin is improved by 272.99%, and the biomass is improved by 94.15%. According to the method, the oxygen-carrying protein VHb is expressed on cytoplasm and organelle, so that the synthesis efficiency and biomass of astaxanthin are remarkably improved. In addition, the intracellular dissolved oxygen level is improved, so that the cost for improving the dissolved oxygen in the fermentation process is reduced.

The application discloses a method for adjusting environment to promote astaxanthin synthesis of Microcystis aeruginosa, and specifically comprises the following steps: culturing Microcystis aeruginosa, determining optimal light quality, determining optimal light intensity and determining optimal temperature and the like. Under the same light intensity and temperature, different light qualities are replaced to culture, white light is taken as a control group, and the optimal light quality for astaxanthin output is determined; under the same light quality and temperature, different light intensities are replaced to culture, and the optimal light intensity for astaxanthin output is determined; under the same light intensity and light quality, different temperatures are replaced to culture, and the optimal temperature for astaxanthin output is determined. In the application, the control variable method is adopted to determine the optimal light quality condition, the optimal light intensity and the optimal temperature for astaxanthin synthesis of Microcystis aeruginosa, so as to provide a theoretical basis for promoting astaxanthin production and resource utilization of Microcystis aeruginosa.

The invention discloses a Yarrowia lipolytica engineering bacterium for producing astaxanthin based on a fusion protein method 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 ketoalcohol enzyme CrtW and beta-carotene hydroxylase CrtZ, and astaxanthin synthesis key enzymes CrtW and CrtZ are subjected to fusion expression by using a short peptide or Linker, so that the astaxanthin is obtained. Meanwhile, the expression of hemoglobin VHb from vitreoscilla is increased; the astaxanthin production performance of the recombinant strain is verified on the basis of protein engineering and fermentation engineering, and the astaxanthin production capacity of the yarrowia lipolytica is further improved.

The invention relates to the field of astaxanthin synthesis, and discloses recombinant saccharomyces cerevisiae as well as a construction method and application thereof. The recombinant saccharomyces cerevisiae provided by the invention comprises the following exogenous genes: coding genes of beta-carotene hydroxylase CrtZ, beta-carotene ketolase CrtW, nicotinamide adenine dinucleotide kinase, ferredoxin-NADPH (nicotinamide adenine dinucleotide phosphate) oxidoreductase and ferredoxin; on the basis of the gene, other related genes are further introduced to realize that CrtZ and CrtW are positioned in lipid droplets by lipid droplet targeting peptide Olesion, so that the yield of astaxanthin is increased, finally geranyl-geranyl diphosphate synthase, phytoene dehydrogenase and phytoene synthase are targeted to endoplasmic reticulum through endoplasmic reticulum targeting peptide, and the yield of astaxanthin is increased. Therefore, the yield of the fermentation tank is up to 440mg / L. The recombinant saccharomyces cerevisiae provided by the invention is clear in genetic background, can stably and efficiently produce astaxanthin, and has a wide application prospect.

The invention relates to an astaxanthin-producing engineering bacterium as well as a construction method and application thereof. According to the astaxanthin-producing engineering bacterium, a crtE gene, a crtI gene, a crtB gene, a crtY gene, a crtZ gene, a crtW gene, a dxs gene and an idi gene are over-expressed. A brand-new genetic modification strategy is designed and comprises the steps that genes in an astaxanthin synthesis route are integrated to an escherichia coli genome, engineering bacteria for producing the astaxanthin are constructed, supply of astaxanthin synthesis precursors is improved by expressing dxs genes and idi genes on the genome, the yield of the astaxanthin is further increased, in addition, a specific crtW gene source is controlled, and the yield of the astaxanthin is increased. According to the present invention, the gene expression quantities of the crtY, the crtZ and the crtW, the astaxanthin yield can be further improved, the maximum shake flask level yield can achieve 27.4 mg / L, and the gene expression on the chromosome is further beneficial to the amplification to the industrial production.

The invention discloses a culture method of all-trans astaxanthin, and relates to the technical field of shrimp culture, and the culture method comprises the following steps: S1, algae species screening and activation; s2, carrying out pre-adaptive domestication culture; s3, gradient stress induction culture; s4, harvesting algae and extracting astaxanthin; according to the culture method, through pre-adaptation domestication-gradient stress induction-dynamic nutrition regulation and control, cell proliferation is effectively promoted, the purity and yield of all-trans astaxanthin are remarkably improved, meanwhile, the cell death rate is effectively reduced, damage of extreme stress to cells is avoided, the algae harvesting efficiency and astaxanthin synthesis stability are guaranteed, and the yield of all-trans astaxanthin is increased. And excellent performance can be presented under different parameter combinations.

The invention relates to the technical field of astaxanthin preparation, and particularly discloses a method for increasing the yield of astaxanthin in haematococcus pluvialis, which comprises the following steps: culturing the haematococcus pluvialis to a growth plateau stage, inoculating the haematococcus pluvialis to a special improved BG-11 culture medium, and carrying out high-light high-salt induction in a columnar photobioreactor; the culture medium is designed for an induction stage and comprises limited sodium nitrate, specific content of salicylic acid, alpha-ketoglutaric acid, sodium chloride and magnesium chloride, and melatonin is not added; in the induction period, step-type increasing illumination intensity is adopted, and white light and ultraviolet light composite light quality treatment is matched. According to the method, the stress tolerance and the astaxanthin synthesis efficiency of cells are remarkably enhanced through the synergistic effect of culture medium components and a light stress procedure, the dry weight of haematococcus pluvialis is 1.69 g L <-1 >, the astaxanthin concentration of haematococcus pluvialis liquid is 105.6 mg L <-1 >, the astaxanthin content of haematococcus pluvialis powder is 6.25% (w / w) within 10 days, the final astaxanthin yield reaches 11.70 mg L <-1 > d <-1 >, and the synchronous great improvement of the astaxanthin yield and the production efficiency is realized.

This invention discloses an engineered astaxanthin-producing probiotic yeast strain, its construction method, and its applications, belonging to the field of synthetic biology. The engineered strain uses β-carotene-producing *Saccharomyces cerevisiae* as the substrate cell. Its genome integrates a fusion gene selected through adaptability screening. This gene is composed of *BDC263crtW* from *Saccharomyces cerevisiae*, a flexible linker peptide, and *AspcrtZ* from *Alcaligenes aeruginosa*, linked sequentially. Simultaneously, based on systematic expression screening of multiple key genes, the endogenous acetyl-CoA synthesis key gene ACS1 was optimized and overexpressed, while the acetic acid metabolism negative regulatory gene YPL062W was knocked out. The construction method includes plasmid construction of the aforementioned fusion expression module and ACS1 expression module, and targeted integration into the genome. This invention significantly improves astaxanthin synthesis levels and enhances the engineered strain's tolerance to the simulated gastrointestinal environment and its antioxidant capacity through enzyme source optimization and substrate channel effects. This engineered strain can be directly used as a functional live bacteria preparation for food and feed additives, with broad prospects for industrial application.

This invention relates to a novel process for producing astaxanthin.

A fusion protein CrtZW and an application in catalyzing a production of astaxanthin from β-carotene are provided, which relates to the field of astaxanthin synthesis technology. The sequence of the fusion protein CrtZW is shown in SEQ ID NO. 4. The fusion protein can catalyze the synthesis of astaxanthin from β-carotene in one step and is suitable for large-scale production.