12 results about "Lycopene cyclase" 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.

The invention belongs to the technical field of enzyme mutants, and discloses a mutant of lycopene cyclase and application of the mutant, glutamic acid at the 321 site of wild type lycopene cyclase which is derived from arabidopsis and has an amino acid sequence as shown in SEQ ID NO.1 is mutated into lysine, phenylalanine at the 319 site of the wild type lycopene cyclase is mutated into leucine, cysteine at the 323 site of the wild type lycopene cyclase is mutated into alanine, and the mutant of the lycopene cyclase is obtained. 1, and the lycopene cyclase mutant with the amino acid sequence as shown in SEQ ID NO. 2 is obtained. BTS1, CrtB and CrtI are integrated into a saccharomyces cerevisiae BY4741 strain, and a chassis strain ZA1 for stably producing alpha-carotene precursor lycopene is constructed; mutant expression plasmids are constructed, positive clone strains are screened out, and the epsilon-cyclization ability of mutant enzymes is remarkably higher than the beta-cyclization ability; the method comprises the following steps: by taking lycopene as a substrate, constructing an expression vector lipid droplet surrounding protein PET10; gene PAH1, DGA1 and Cat2 related to TAG synthesis and perilipid droplet protein PET10 are constructed on an expression vector and converted into ZA1, and when lycopene is used as a substrate, the efficiency of catalytic production of alpha-carotene is improved.

The invention provides recombinant saccharomyces cerevisiae for producing beta-carotene and application of the recombinant saccharomyces cerevisiae, and belongs to the field of genetic engineering. According to the invention, a beta-carotene metabolic pathway is constructed in saccharomyces cerevisiae, screening is carried out on a key gene lycopene cyclase gene in the metabolic pathway, and site-directed mutagenesis is carried out on lycopene cyclase derived from Rhizomucor circinelloides f. Lusitanius, so that a mutant is constructed. According to the invention, the constructed mutant is expressed in saccharomyces cerevisiae, so that the titer of beta-carotene is remarkably improved.

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 firstly provides an enzyme system for improving the yield of astaxanthin. The enzyme system comprises beta-carotene hydroxylase crtZ, beta-carotene ketolase crtW and lycopene cyclase crtY which are found by the inventor and have remarkably improved conversion rate. The beta-carotene hydroxylase crtZ has an amino acid sequence as shown in SEQ ID NO: 2, the beta-carotene ketolase crtW has an amino acid sequence as shown in SEQ ID NO: 3, and the lycopene cyclase crtY has an amino acid sequence as shown in SEQ ID NO: 1. On the basis, coding genes of the beta-carotene hydroxylase crtZ, the beta-carotene ketolase crtW and the lycopene cyclase crtY are over-expressed in a basic strain for producing the lycopene, and an escherichia coli recombinant strain for producing the astaxanthin at high yield is constructed. When the recombinant escherichia coli is used for producing astaxanthin through fermentation, the yield and the conversion rate of the astaxanthin are obviously improved; according to the present invention, glycerol is adopted as a substrate, and fermentation is performed in a 5 L fermentation tank for 60 h to obtain 1.54 g / L of astaxanthin (1.18 g / L in the prior art), such that the technical problem of low yield of astaxanthin produced through microbial fermentation in the prior art is solved, the foundation is laid for the industrial production of astaxanthin, and wide application prospects and huge market values are provided.

The invention relates to a recombinant yarrowia lipolytica strain with high yield of astaxanthin as well as a construction method and application thereof. The preparation method comprises the following steps: expressing geranyl geranyl diphosphate synthase CrtE, phytoene synthetase / lycopene cyclase CrtYB, phytoene desaturase CrtI, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR, beta-carotene ketolase CrtW, beta-carotene hydroxylase CrtZ, ATP (adenosine triphosphate) citrate lyase ACL, acetyl-CoA synthase ACS, choline kinase CK and inositol polyphosphate kinase IPK in host bacteria, so as to obtain a recombinant bacterium; the expression cassettes of farnesyl diphosphate synthase ERG20 and isopentenyl pyrophosphate isomerase IDI are obtained. Meanwhile, a photocatalytic material O-g-C3N4 / PEI is added to receive visible light excitation and generate reducing electrons, synthesis of intracellular NADPH is promoted, and then accumulation of astaxanthin is promoted. And continuous fed-batch fermentation is carried out in a 5L fermentation tank, so that the production performance of yarrowia lipolytica is greatly improved, and a foundation is laid for industrial production of high-value terpenoids.

Recombinant microorganisms configured for enhanced production of carotenoids and other compounds and methods of using the recombinant microorganisms for the production of same. The recombinant microorganism can include a modification that decreases phytoene synthase (CrtB) activity, a modification that decreases lycopene-forming phytoene desaturase (CrtI) activity, a modification that decreases lycopene cyclase (CrtY) activity, a modification that decreases beta-carotene hydroxylase (CrtZ) activity, a modification that decreases 2,2′-beta hydroxylase (CrtG) activity, and / or a modification that increases beta-carotene ketolase (CrtW) activity. The recombinant microorganism can be from the genus Novosphingobium, such as Novosphingobium aromaticivorans.

This invention discloses an engineered strain of *Yersinia lipolytica* for synthesizing astaxanthin, its preparation method and application, a method for producing astaxanthin, and a method for increasing astaxanthin yield. The engineered strain of *Yersinia lipolytica* for synthesizing astaxanthin is obtained by expressing phytoene dehydrogenase CarB, phytoene synthase / lycopene cyclase CarRP, β-carotene ketolase CrtW, β-carotene hydroxylase CrtZ, and ferricoxane reductase RFNR in a host strain. RFNR Derived from Chlamydomonas rhinelandii Chlamydomonas reinhardtii Haematococcus pluvialis Haematococcus pluvialis This strain, through heterologous expression of RFNR, efficiently provides reducing power to CrtZ, significantly enhancing the catalytic activity of CrtZ and ultimately achieving higher astaxanthin yield.

The invention discloses a genetically engineered bacterium for improving rate-limiting enzyme to promote xanthophyll synthesis and application of the genetically engineered bacterium, and belongs to the technical field of genetic engineering. According to the genetically engineered bacterium, a lycopene-producing strain is taken as a starting bacterium, and coding genes of a lycopene epsilon-cyclase mutant, lycopene beta-cyclase, a carotene beta-hydroxylase mutant and carotene epsilon-hydroxylase are extrinsic introduced; according to the invention, a lycopene epsilon-cyclase mutant with higher enzymatic activity and / or a carotene beta-hydroxylase mutant with higher enzymatic activity are / is utilized to improve the limitation of a rate-limiting enzyme in the lutein synthesis process, and further, the lutein synthesis route is subjected to compartment regulation and control; and the lycopene epsilon-cyclase and / or the lycopene beta-cyclase are / is positioned in the intracellular lipid droplets, so that the synthesis of the lutein is comprehensively enhanced.

The invention provides recombinant yarrowia lipolytica for co-production of microbial oil and astaxanthin as well as a construction method and application of the recombinant yarrowia lipolytica, and belongs to the technical field of genetic engineering. The recombinant yarrowia lipolytica expresses geranyl geranyl diphosphate synthase CrtE derived from phaffia rhodozyma, phytoene synthetase / lycopene cyclase CrtYB, phytoene desaturase CrtI, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR derived from saccharomyces cerevisiae, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR derived from saccharomyces cerevisiae, 3-hydroxy-3-methylglutaryl CoA reductase tHMGR derived from saccharomyces cerevisiae, and 3-hydroxy-3-methylglutaryl CoA reductase tHMGR derived from saccharomyces cerevisiae. The beta-carotene ketolase CrtW and the beta-carotene hydroxylase CrtZ are derived from the haematococcus pluvialis. According to the method, coproduction of the microbial oil and the astaxanthin by using the yarrowia lipolytica is realized by using the molasses as a cheap raw material in a 5L fermentation tank, the yield of the microbial oil reaches 80.14% and the yield of the astaxanthin reaches 3.09 g / L after fermentation is performed for 168 hours, and a foundation is laid for further industrialization.

This invention belongs to the field of bioengineering technology, specifically relating to a recombinant yeast strain that utilizes yeast as a host cell and achieves efficient astaxanthin biosynthesis through metabolic engineering. The invention further discloses its construction method and applications. The recombinant yeast strain of this invention is obtained by introducing expression cassettes of geranylgeranyl diphosphate synthase (GGPPS), phytoene synthase / lycopene cyclase (CrtYB), phytoene desaturase (CrtI), 3-hydroxy-3-methylglutaryl-CoA reductase (HMG2), β-carotene ketolase (CrtW), and β-carotene hydroxylase (CrtZ) into *Saccharomyces cerevisiae* and dynamically regulating the expression of the ERG20 gene. The modified recombinant *Saccharomyces cerevisiae* strain of this invention can efficiently synthesize astaxanthin, achieving the efficient synthesis of the natural product astaxanthin in *Saccharomyces cerevisiae*.

The invention relates to the technical field of genetic engineering and biosynthesis, in particular to a lycopene cyclase variant and application of the lycopene cyclase variant in production of alpha-carotene. The present invention provides a lycopene bifunctional cyclase variant, which catalyzes the synthesis of alpha-carotene only from lycopene without producing beta-carotene, as well as a gene encoding the variant. The invention provides a yarrowia lipolytica genetically engineered bacterium for producing alpha-carotene by using the lycopene bifunctional cyclase variant as well as a construction method and application of the yarrowia lipolytica genetically engineered bacterium, and the constructed yarrowia lipolytica genetically engineered bacterium can preferentially catalyze and synthesize lycopene into alpha-carotene instead of beta-carotene. The content of the alpha-carotenoid in the fermentation bacteria liquid is 85% or above of the total carotenoid, and a technology and thought are provided for industrial production of the natural alpha-carotenoid through a biosynthesis method.