52 results about "Carotenoid synthesis" patented technology

The present invention provides engineered cells and methods for utilizing same. Methods of enhanced carotenoid synthesis utilizing cells genetically disrupted for a yjiD, hnr or yjfP gene, or further disrupted for a gdhA, gpmB, aceE, ppc, talB or fdhF gene, or any combination thereof, or cells inhibited for their expression, activity or function are disclosed. Additionally, methods of enhanced carotenoid synthesis utilizing cells genetically disrupted for gdhA, aceE, fdhF, yjiD, hnr or yjfP gene expression or any combination thereof and ackA, appY, aspC, clp, clpP, clpXP, crcB, csdA, cyaA, evgS, fdhA, fdhD, feoB, funA, glnE, glxR, gntK, hycI, lipB, lysU, modA, moeA, nadA, nuoC, nuoK, pflB, pitA, pst, pstC, pta, p-yjiD, sohA, sipA, yagR, yaiD, ybaS, ycfZ, ydeN, yebB, yedN, yfcC, ygjP, yibD, yjfP, yjhH, or yliE gene expression, or a combination thereof or cells inhibited for their expression, activity or function are disclosed. Methods of enhanced carotenoid synthesis utilizing cells genetically engineered to overexpress dxs, idi, ispFD, yjiD, rpoS, torC, appY, ydgK, yeiA, yedR, tort, arcB, yggT, purDH, yfjN or a combination thereof, or further disrupted for the above-referenced genes are disclosed. Methods for identifying genes involved in optimized production of a carotenoid, and cells disrupted for, or inhibited for the expression, activity or function of genes thus identified are described.

The invention discloses a tobacco carotenoid isomerase gene. The base sequence of the tobacco carotenoid isomerase gene is represented by SEQIDNO:1. In tobacco, a key carotenoid synthesis gene NtCRTISO is cloned, and the gene can be inhibited to effectively improve the carotenoid content of tobacco leaf and improve the photoinhibition resistance of tobacco plants. The invention discloses an important function of the CRTISO gene in tobacco in carotenoid synthesis. The CRTISO gene plays a great role in the regulation of the composition of carotenoid. A method for improving the content of carotenoid, especially beta-carotene, in the tobacco is searched, and a good theoretic guidance is provided for the improvement of the molecules of the plants.

The invention discloses an acetyl coenzyme A acetyltransferase gene RKAcaT2 and application thereof. The nucleotide sequence of the gene is shown in SEQ ID NO:1, the amino acid sequence encoded by thegene is shown in SEQ ID NO:2, the gene is a key enzyme gene synthesized by carotenoid in Rhodosporidium kratochvilovae YM25235 and has the functions of the acetyl coenzyme A acetyltransferase, and the carotenoid produced by the Rhodosporidium kratochvilovae YM25235 can be controlled; microorganisms are transformed through the genetic engineering means so as to increase the yield of carotenoid inthe microorganisms, and a foundation is laid for large-scale commercial production of the carotenoid.

The invention discloses an HMG-CoA synthetase gene RKHMGCS, the nucleotide sequence is shown as the SEQ ID NO:1, and the amino acid sequence of the gene coding is shown as the SEQ ID NO:2; the gene isthe key enzyme gene for synthesis of the carotenoid in the Rhodosporidium kratochvilovae YM 25235, the gene has the HMG-CoA synthetase function, and the Rhodosporidium kratochvilovae YM 25235 can becontrolled to produce the carotenoid; the microorganism is modified by the means of genetic engineering, the yield of the carotenoid in the microorganism body is improved, and a foundation is laid forlarge-scale commercialized production of the carotenoid.

The invention relates to an application of a sll0147 gene in synthesizing synechocystis carotenoids, wherein the nucleotide sequence of the sll0147 gene in the application is shown as SEQ ID NO.1. For the first time, the invention discloses the important effect of the sll0147 gene of synechocystis PCC6803 in synthetising carotenoids; by increasing the expression of the sll0147 gene in the synechocystis PCC 6803 through an overexpression method by the applicant, the percentage composition of the carotenoid component in a mutant strain is obviously changed, wherein the content of myxoxanthophyll is increased by 47.4%, the content of zeaxanthine is increased by 93.8%, while the content of echinenone is reduced by 60.9%, and the content of beta-carotene is reduced by 15.9%.

The invention discloses a phytoene dehydrogenase mutant gene and application thereof in carotenoid synthesis. The mutant gene is obtained by performing multiple rounds of mutation on a phytoene dehydrogenase coding gene by means of iterative orthogenesis and through a high-throughput screening method based on bacterial colony color change. When the phytoene dehydrogenase mutant gene is used to synthesize lycopene, A178S mutant is obtained in the first round of orthogenesis, and lycopene yield is increased by 27.3%; A178S / N280T mutant and A178S / M234I mutant are obtained by using the A178S mutant as a base for orthogenesis, and lycopene yield is further increased by 51.4% and 18.3% while the lycopene yield is not further increased by combining A178S, N280T and M234I; A178S / N280T / E239P mutantis obtained by performing orthogenesis on the A178S / N280T mutant, and A178S / N280T / E239P mutant is finally obtained through three rounds of orthogenesis and screening.

The present invention provides genetically manipulated cells and methods for utilizing same. Methods of enhanced carotenoid synthesis utilizing cells genetically disrupted for a yjiD, hnr or yjfP gene, or further disrupted for a gdhA, gpmB aceE, ppc, talB or fdhF gene, or any combination thereof, or cells inhibited for their expression, activity or function are disclosed. Methods of enhanced carotenoid synthesis utilizing cells genetically engineered to overexpress dxs, idi, yjiD, rpoS, torC, appY, ydgK, yeiA, yedR, tort, arcB, yggT, purDH, yfjN or a combination thereof, or further disrupted for the above-referenced genes are disclosed. Methods for identifying genes involved in optimized production of a carotenoid, and cells disrupted for, or inhibited for the expression, activity or function of genes thus identified are described.

The invention discloses a 3-hydroxy-3-methylglutaryl coenzyme A reductase gene RKHMGR and application thereof. A nucleotide sequence is as shown in SEQ ID NO: 1, an amino acid sequence encoded by thegene is as shown in SEQ ID NO: 2, the gene is a key enzyme gene of carotenoid synthesis in YM25235 of Rhodosporidium Kratochvilovae, is the first rate limiting enzyme in an MVA pathway, and can regulate the YM25235 of the Rhodosporidium Kratochvilovae to generate the carotenoid. According to the 3-hydroxy-3-methylglutaryl coenzyme A reductase gene RKHMGR and the application thereof, microorganismis transformed by means of genetic engineering, the yield of the carotenoid in the microorganism is improved, and a foundation is laid for large-scale commercial production of the carotenoid.

The present invention provides engineered cells and methods for utilizing same. Methods of enhanced carotenoid synthesis utilizing cells genetically disrupted for a yjiD, hnr or yjfP gene, or further disrupted for a gdhA, gpmB, aceE, ppc, talB or fdhF gene, or any combination thereof, or cells inhibited for their expression, activity or function are disclosed. Additionally, methods of enhanced carotenoid synthesis utilizing cells genetically disrupted for gdhA, aceE, fdhF, yjiD, hnr or yjfP gene expression or any combination thereof and ackA, appY, aspC, clp, clpP, clpXP, crcB, csdA, cyaA, evgS, fdhA, fdhD, feoB, funA, glnE, glxR, gntK, hycI, lipB, lysU, modA, moeA, nadA, nuoC, nuoK, pflB, pitA, pst, pstC, pta, p-yjiD, sohA, stpA, yagR, yaiD, ybaS, ycfZ, ydeN, yebB, yedN, yfcC, ygjP, yibD, yjfP, yjhH, or yliE gene expression, or a combination thereof or cells inhibited for their expression, activity or function are disclosed. Methods of enhanced carotenoid synthesis utilizing cells genetically engineered to overexpress dxs, idi, ispFD, yjiD, rpoS, torC, appY, ydgK, yeiA, yedR, tort, arcB, yggT, purDH, yfjN or a combination thereof, or further disrupted for the above-referenced genes are disclosed. Methods for identifying genes involved in optimized production of a carotenoid, and cells disrupted for, or inhibited for the expression, activity or function of genes thus identified are described.

The invention provides a mutant chlorella strain capable of realizing high yield of zeaxanthine and other carotenoids essential to the human body, and a culturing method thereof. The culturing method comprises the following steps: subjecting Chlorella zofingiensis ATCC 30412 to nitrosoguanidine mutagenesis; then coating a Kuhl solid medium containing DPA with treated chlorella cells for culture; selecting a single chlorella colony from the Kuhl solid medium and culturing the single chlorella colony on a Kuhl liquid medium; adding glucose in the middle and later periods of logarithm for induction of carotenoid synthesis; and detecting the variety changes and content of carotenoids in the obtained chlorella body so as to obtain the strain which can realize high yield of zeaxanthine, xanthophyll and beta-carotene and is named as CZ-ZEA1. Under the induction condition that the concentration of glucose is 30 g / L, the biomass of the CZ-ZEA1 is 12.95 g / L, and the contents of zeaxanthine, xanthophyll and beta-carotene reach 2.176 mg / g, 1.10 mg / g and 1.211 mg / g, respecitively. The mutant chlorella strain is an edible alga species and contains high-content optimally-proportioned carotenoids essential to the human body, especially zeaxanthine rare in nature; so the mutant chlorella strain is an ideal new-resource functional foodstuff for preventing deterioration and lesion of the eyes of middle aged and old people and has good development and application prospects.

The invention relates to recombinant bacillus subtilis capable of synthesizing carotenoid 4,4'-diaponeurosporene with 30 carbon atoms (C30), and belongs to the biotechnology genetic engineering field. A bacillus subtilis expression plasmid pMK3-crtmn (which is capable of expressing C30 carotenoid synthases Crtm and Crtn in bacillus subtilis) containing C30 carotenoid synthase genes crtm and crtn is successfully constructed, and is successfully electro-transformed into bacillus subtilis WB800. The WB800 strain after transformation turns from the original color of white to yellow. A pigment component extracted from the recombinant WB800 is identified as 4,4'-diaponeurosporene by high performance liquid chromatography (HPLC). 4,4'-diaponeurosporene can stimulate maturation of dendritic cells, and generated cytokines (IL-6, IL-10, IL-12p70 and TNF[alpha]) have the amount increased, and moreover and have stronger ability to stimulate proliferation of T lymphocytes. With oral administration of the bacillus subtilis capable of synthesizing 4,4'-diaponeurosporene, the dextran sulphate sodium (DSS)-induced mice colitis symptoms can be significantly reduced. The bacillus subtilis capable of producing 4,4'-diaponeurosporene is expected to be developed as probiotics for prevention of colitis.

The invention discloses a 3-ketoacyl coenzyme A thiolase gene RKACAA1-2, the nucleotide sequence of the 3-ketoacyl coenzyme A thiolase gene RKACAA1-2 is shown as SEQ ID NO: 1, and the amino acid sequence coded by the gene is shown as SEQ ID NO: 2; the gene is separated from rhodosporidium kratochvilovae YM25235, the gene is transferred into the rhodosporidium kratochvilovae YM25235 through transformation, and the experimental result shows that the overexpression of the RKACAA1-2 gene can cause the improvement of the carotenoid synthesis level of the YM25235 strain; according to the gene, the microorganisms are modified by means of genetic engineering, so that the yield of carotenoid in the microorganisms is increased, and a foundation is laid for large-scale commercial production of carotenoid.

The invention discloses astaxanthin synthetase of sphingomonas, an encoding gene of astaxanthin synthetase and a method for genetic manipulation of sphingomonas. The invention verifies that the sphingomonas can be used for producing astaxanthin, a biological synthesis way capable of producing the astaxanthin is contained, an MEP way and a carotenoid synthesis way are used for producing the astaxanthin in the sphingomonas, and the fact that in the sphingomonas, crtE and crtZ having relatively low homology with the known genes have the function of synthesizing the astaxanthin, can be further proven. Both the enzyme and the encoding gene thereof required by the astaxanthin biosynthetic way of the sphingomonas are not reported in the prior art, so that more resources are provided to the biosynthetic metabolism and transformation of the carotenoid. Genetic manipulation can be carried out on the sphingomonas by taking pBBR1MCS2 as a carrier and taking EGFP as a reporter gene, so that a foundation is laid for the genetic modification of the environmental microorganism of the type and the finding of the degradation mechanisms.

The invention discloses a carotenoid synthetic route Tagetes erecta violaxanthin de-epoxidase (TeVDE1) gene. The TeVDE1 is characterized in that the nucleotide sequence of the TeVDE1 gene is shown as SEQ ID NO: 1 in a sequence table, the amino acid sequence of the TeVDE1 gene is shown as SEQ ID NO: 2 in the sequence table. The TeVDE1 gene is obtained from tagetes erecta petal tissue in a clone mode. Real-time quantification analysis and a xanthophyll content determination result show that the expression quantity of the TeVDE1 gene in Tagetes erecta flower buds is high, and the expression level of the TeVDE1 is related to the xanthophyll content. The SEQ ID NO: 1 gene has an effect on xanthophyll synthetic accumulation, and the TeVDE1 obtained from the pigment tagetes erecta in a clone mode can be used for increasing the pigment contents of xanthophylls and other carotenoid.

The invention discloses a method for detecting the synthesis capability of carotenoid in a fruit. According to the method, lycopene cyclase inhibitor, namely CPTA, is adopted for treating the fruit to be detected, degradation of the carotenoid in the fruit is blocked so that the carotenoid can be accumulated in the form of lycopene, and the capability of the fruit for synthesizing the carotenoid can be evaluated objectively by detecting the lycopene. The method is generally applicable to various fruits such as peaches, apricots and apples.

The invention discloses a fermentation medium beneficial to synthesis of saccharomyces cerevisiae carotenoid. The culture medium comprises a saccharomyces cerevisiae basic culture medium and copper ions. The fermentation medium provided by the invention significantly improves the accumulation of recombinant saccharomyces cerevisiae carotenoid (by 10 times). According to the invention, transcriptomics reveals that the addition of zinc ions and copper ions in the culture medium can significantly promote the synthesis of the carotenoid, and the zinc ions, copper ions and the carotenoid have a synergistic enhancement effect.

The invention discloses a research method for the synthetic process of haematococcus pluvialis astaxanthin based on an infrared spectroscopy microscopic imaging technology. The method comprises the following steps that inoculation and culture are carried out on haematococcus pluvialis; an inhibitor performs treatment on haematococcus pluvialis strains; the infrared spectroscopy of the astaxanthin,beta-carotene and lycopene are collected; the infrared microscopic spectroscopy of haematococcus pluvialis cells are detected; and comparison and analysis of imaging of the microscopic infrared spectroscopy are carried out. According to the method, the infrared microscopic spectroscopy imaging technology is utilized, the haematococcus pluvialis cells are treated by combining the inhibitor synthesized by carotenoid, and high-spatial-resolution in-situ observation is carried out on the contents of various components of the haematococcus pluvialis cells in the astaxanthi accumulation process, sothat the effects of the components in the synthesis path of the astaxanthin can be conveniently analyzed. The method has important significance in research of the synthesis path of the carotenoid inthe haematococcus pluvialis cells and large-scale breeding of the haematococcus pluvialis for production of the astaxanthin.

The invention discloses an application of complexing agents in enhancing ornamental quality of rye grass with an aim to enhance growth of seedlings and synthesis of chlorophyll and carotenoids. The complexing agents refer to GLDA (tetrasodium glutamate diacetate) and IDS with the concentration of 6-9mmol kg<1>; the complexing agents GLDA and IDS with respective concentration of 9mmol kg<1> are used for increasing total chlorophyll and carotenoid content; the complexing agents GLDA and IDS with respectively concentration of 6mmo kg<1> are used for increasing the total chlorophyll and carotenoid content; the complexing agents of GLDA and IDS with concentration of 3mmol kg<1> are used for increasing total chlorophyll and carotenoid content. The GLDA or IDS different in concentration are added to a turf medium, and enhanced regulation on synthesis of the GLDA or IDS on the chlorophyll and carotenoid of the rye grass is realized; technical support for green quality of the rye grass can be further improved.

The invention discloses a gene RKcrtYB. The gene is isolated from rhodosporidium kratochvilovae YM25235. The nucleotide sequence of the gene is as shown in SEQ ID NO:1, and the amino acid sequence encoded by the gene is as shown as EDQ ID NO:2. In the present invention, the gene is linked into a vector and transferred into the YM25235, and the translated protein is involved in the synthesis of beta-carotenoids. The gene can enable the rhodospora erythraea YM25235 to improve the synthesis of the beta-carotenoids, and lays a foundation for the large-scale commercial production of the beta-carotenoids.

The invention belongs to the technical field of gene engineering, and discloses a CsMYB110 gene and an application thereof in regulation of synthesis of carotenoids. The nucleotide sequence of the CsMYB110 gene is as shown in SEQ ID NO. 1 in a sequence table. The amino acid sequence of protein coded by the CsMYB110 gene is as shown in SEQ ID NO. 2 in the sequence table. The expression mode of CsMYB110 is highly related to synthesis of the carotenoids in tea tree leaves, and the gene can significantly promote synthesis and accumulation of the carotenoids in plants. Cloning of the gene not only facilitates analysis of a regulation mechanism of the carotenoids in tea tree leaves, but also is good for cultivation of tea tree varieties with high carotenoid content and improvement of tea quality, and great application value is achieved.

A microorganism is provided which allows carotenoid production in industrial production scale. A method of preparing a carotenoid includes: culturing a cell transformed with a DNA chain having a DNA sequence selected from the group consisting of the following (a) to (f) or a cell transformed with a vector having a DNA sequence selected from the group consisting of the following (a) to (f) in an appropriate culture condition, and isolating carotenoid from the cell or a culture medium: (a) DNA sequence encoding a polypeptide having such an enzymatic activity of Paracoccus sp. Strain MBIC1143 that converts a methylene group at 4 position in β-ionone ring into a keto group (crtW), described in SEQ ID NO: 2, or a substantially homologous DNA sequence thereof; (b) DNA sequence encoding a polypeptide having such an enzymatic activity of Paracoccus sp. Strain MBIC1143 that adds one hydroxyl group to a carbon at 3-position of 4-keto-β-ionone ring and / or at 3-position of β-ionone ring (crtZ), described in SEQ ID NO: 3, or a substantially homologous DNA sequence thereof; (c) DNA sequence encoding a polypeptide having such an enzymatic activity of Paracoccus sp. Strain MBIC1143 that converts lycopene into β-carotene (crtY), described in SEQ ID NO: 4, or a substantially homologous DNA sequence thereof; (d) DNA sequence encoding a polypeptide having such an enzymatic activity of Paracoccus sp. Strain MBIC1143 that converts phytoene into lycopene (crtI), described in SEQ ID NO: 5, or a substantially homologous DNA sequence thereof; (e) DNA sequence encoding a polypeptide having prephytoene synthase activity of Paracoccus sp. Strain MBIC1143 (crtB), as described in SEQ ID NO: 6, or a substantially homologous DNA sequence thereof; and (f) DNA sequence encoding a polypeptide having geranylgeranyl diphosphate synthase activity of Paracoccus sp. Strain MBIC1143 (crtE), as described in SEQ ID NO: 7, or a substantially homologous DNA sequence thereof.

The invention discloses a method for producing carotenoids by using food organic wastewater, relating to a method for producing carotenoids by using organic wastewater. By adopting the method, the problems that in an existing method for synthesizing carotenoids by using PSB (photosynthetic bacteria) to treat food organic wastewater, the capacity of PSB for degrading organic matters is limited, relatively less substrates for synthesizing carotenoids are provided, the yield of bacterial carotenoids in wastewater treatment is low, and the recycling process of wastewater treatment is blocked can be solved. The method comprises the following steps: firstly, adjusting the pH value of the food organic wastewater, and then adding rhodobacter sphaeroides and growth-promoting bacteria; secondly, performing treatment for 72-96h under micro-aerobic conditions; and thirdly, recycling and extracting to obtain carotenoids. Compared with an existing conventional technique for treating the food organic wastewater by independently using PSB, the bacterium yield and carotenoid yield can be increased by more than 25% and 100% respectively by adopting the method disclosed by the invention, and the removal rate of COD in the food organic wastewater also can be increased by more than 78%.

A PSY2 gene is a key gene participating in carotenoid synthesis. A new PSY gene DcPSY2 is cloned from carrots. The open reading frame length of the gene is 1317bp, and 438 amino acids are encoded. Theprotein encoded by DcPSY2 belongs to hydrophilic protein. The protein is closest to the evolutionary relationship of leguminous rosewood PSY and has high conservative property. Fluorescent quantitative expression analysis shows that the relative expression level of the DcPSY2 gene reaches a peak value after treatment for 2h under high temperature and high salt stress, and the relative expressionlevel is significantly different from that of a control group (P is less than 0.05). Under low temperature and drought stress, the expression level of the DcPSY2 gene of the carrots is increased mostobviously after treatment for 1h. The DcPSY2 gene of the carrots is cloned from the carrots by using a polymerase amplification technology, and the DcPSY2 gene of the carrots responds to the adversitystress of carrot plants.

The invention discloses saccharomyces cerevisiae bred by a space breeding technology and mutation site application thereof. The saccharomyces cerevisiae W2 is preserved in Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC No: 61335. The strain W2-A-5 is obtained through further modification on the basis of W2, and the preservation number of the strain W2-A-5 is GDMCC No:61337. The saccharomyces cerevisiae mutant strain W2 is obtained through microgravity mutagenesis screening, and the carotenoid content of the strain can reach 45 mg / g of dry cell weight under the shake flask fermentation condition. W2-A-5 is obtained through further transformation on the basis of the strain, and the content of carotenoid can reach 68 mg / g of dry cell weight. Through genome sequencing and reverse metabolic engineering research, CHO2 gene inactivation is determined to be a main factor for improving the synthesis capability of the W2 carotenoid. After CHO2 in the strain without growth retardation is inactivated, the carotenoid synthesis capability is remarkably improved, and a new modification target is provided for improving the carotenoid synthesis capability by directionally modifying the saccharomyces cerevisiae.

The invention belongs to the technical field of microbial fermentation, and particularly belongs to a method for increasing the yield of beta-carotene produced by fermentation of Blakeslea trispora. When Blakeslea trispora is used for liquid submerged fermentation, an ethephon solution with a certain concentration is added, ethephon slowly releases ethylene, and ethylene plays a role in promotingcarotenoid synthesis related genes on the transcription level, so that accumulation of a large amount of beta-carotene is promoted, and the fermentation yield of beta-carotene is increased. The methodis low in cost, simple and convenient to operate and obvious in effect, and can replace various fermentation accelerators such as an oxygen carrier, a surfactant, a precursor substance and the like to increase the yield of beta-carotene, so that other fermentation accelerators are prevented from being too high in price and poison to strains, and a good basis is provided for industrial production.Compared with a blank control group, the yield of beta-carotene is increased by 54%-78%, and meanwhile, the fermentation period is shortened to 3 days from the original 5 days and is shortened by 40%.

The invention discloses a crgA gene of Blakeslea trispora negative bacteria as well as a gene cloning method and application of the crgA gene and belongs to the technical field of genetic engineering. The gene cloning method disclosed by the invention comprises the following steps: designing four primer pairs according to a crgA gene sequence of Blakeslea trispora positive bacteria and successfully cloning to obtain the crgA gene of the Blakeslea trispora negative bacteria; carrying out homologous alignment analysis on a crgA gene of the Blakeslea trispora positive bacteria and the crgA gene of the Blakeslea trispora negative bacteria; then knocking out the gene, and carrying out comparative analysis a gene knock-out plant on a wild plant in aspects of phenotypic characteristic, key enzyme gene transcription level, carotenoid synthesis level and the like to initially display the effect of the gene as a negative regulatory factor in Blakeslea trispora. According to the crgA gene of the Blakeslea trispora negative bacteria as well as the gene cloning method and application of the crgA gene, disclosed by the invention, the crgA gene of the Blakeslea trispora negative bacteria is cloned by a gene engineering technology, which is beneficial to analyzing a regulation and control mechanism of the crgA gene in the process of producing carotenoid by the Blakeslea trispora and improving the yield of the carotenoid; the crgA gene has a good application prospect.

The invention belongs to the technical field of plant gene engineering, and in particular, relates to an expression vector for comprehensive down-regulation of cotton lycopene cyclase (Lyc) genes and an application thereof. The technical scheme comprises that coding sequences selected from 5 pairs of cotton Lyc genes respectively construct an RNAi element in series; the element is transcribed in cotton by a transgenic method, and ultimately, the purposes of down-regulating the 5 pairs of Lyc genes and regulating synthesis of carotenoids in the cotton simultaneously are achieved. The RNAi element and the vector can be used for regulating the content and type of the carotenoids in the cotton.