24 results about "Elongase" patented technology

The invention discloses a schizochytrium limacinum genetic engineering strain capable of remodeling an FAS (Fibroblast Activated Sequence) pathway as well as a construction method and application thereof. The schizochytrium limacinum gene engineering strain is obtained by heterologous expression of a C16 elongase gene E16, a C18 elongase gene E18, a delta 9 desaturase gene D9, a delta 5 desaturase gene D5, a delta 6 desaturase gene D6 and a delta 15 desaturase gene D15 in a schizochytrium limacinum wild type. The recombinant schizochytrium limacinum described herein is capable of producing EPA comprising 2.08% by total fatty acids while also producing DHA comprising at least more than 41% by total fatty acids. In addition, the invention discloses a method for producing unsaturated fatty acid through a double-path channel of PKS and FAS, which is beneficial to promotion and application of a technology for producing EPA and DHA by schizochytrium limacinum.

The invention belongs to the technical field of biology, and discloses a yarrowia lipolytica engineering strain for efficiently producing eicosapentaenoic acid based on citric acid metabolism regulation, a method and application of the yarrowia lipolytica engineering strain. Delta9 elongase, Delta8 desaturase, Delta5 desaturase and Delta17 desaturase genes in a fatty acid synthesis route, heterologous ATP-citrate lyase and a mitochondrial citric acid vector are overexpressed. According to the invention, ATP-citric acid lyase and a mitochondrial citric acid carrier gene which are respectively derived from Schizochylene sp. And Saccharomyces cerevisiae are jointly expressed in the yarrowia lipolytica engineering strain, so that the yields of EPA and grease in the yarrowia lipolytica engineering strain are remarkably improved, and compared with a control strain, the yields of the grease and the EPA are respectively improved by 243.35% and 161.85%.

The invention belongs to the technical field of biology, and discloses an exogenous cofactor enhanced yarrowia lipolytica strain, a method and application in efficient synthesis of EPA (eicosapentaenoic acid) and lipids.The chassis strain of the yarrowia lipolytica strain is yarrowia lipolytica Po1f, desaturase is partially prolonged in an overexpression fatty acid synthesis route in the strain, and the EPA and the lipids.The yield of the EPA and the lipids.The yield of the EPA and the lipids.The yield of the EPA and the lipids. The desaturase gene mainly comprises a delta 9 elongase gene, a delta 8 desaturase gene, a delta 5 desaturase gene and a delta 17 desaturase gene, and G6PDH and malic enzyme are overexpressed at the same time on the basis. The strain provided by the invention improves the EPA and lipid yield of yarrowia lipolytica; the method is suitable for constructing a genetic engineering strain with high yield of EPA and lipid, provides a basis for producing grease rich in EPA from yarrowia lipolytica, and solves the problems of non-sustainability, high cost and the like caused by obtaining EPA from fish.

The invention provides a BFD mutant and application thereof as a carbon chain elongase, and the BFD mutant is characterized in that (a) compared with an amino acid sequence as shown in SEQ ID NO.1, the BFD mutant contains at least one of the following sites: L43, A73, T100 and K165, and an amino acid sequence which is subjected to amino acid mutation and has an active function of the carbon chain elongase; or (b) an amino acid sequence which is obtained by substitution and / or deletion and / or addition of one or more conserved amino acid residues of the BFD mutant and the amino acid sequence shown in (a) and has the same function. The enzyme activity of the BFD mutant is obviously improved, and the yield and conversion rate of glycolaldehyde and / or 1, 3-dihydroxyacetone generated from formaldehyde are improved.

The invention relates to a method for forming a hair follicle-like structure by co-culturing an immortalized hair papilla cell line derived from human hair follicles and keratinocytes. Specifically disclosed is a hair papilla cell line (SV40T-hTERT DPC) which is immortalized by forced expression of SV40T, which deactivates a cancer suppressor gene, in human hair follicle-derived hair papilla cells and hTERT gene, which is a telomerase elongase promoting gene, and keratinocytes, are co-cultured three-dimensionally. From the result, it has been confirmed that a hair follicle-like structure is formed in which a cuticle grows so that a hair follicle analogue in combination with a primary-cuticle hair follicle-derived cell has a shape similar to that of the primary-cuticle hair follicle-derived cell. Therefore, the method can be applied to multiple fields of animal replacement experiments, efficacy evaluation of materials for preventing alopecia / promoting hair follicle growth and the like. Through immortalized hair papilla cells, higher result consistency can be ensured compared with that in the prior art, and stable hair follicle-derived cell supply can be realized, so that the problem of high entry barriers of efficacy evaluation of alopecia materials based on cells is expected to be solved.

The invention discloses a method for quickly constructing a nucleic acid library, and belongs to the technical field of library construction. A DNA terminal repair-linker connection reagent combination is provided, a 3'protruding end of a DNA fragment is cut off and / or a 5 'protruding end of the DNA fragment is filled in through a terminal repair enzyme so as to finally form a double-chain DNA fragment with a blunt terminal, and the DNA fragment subjected to terminal repair is connected with a linker through a ligase. According to the method, a complementary chain of a connector connected with a DNA fragment is extended through nucleic acid elongase, so that the DNA fragment with double-chain connectors at the two ends is obtained, tail end repair and connector connection are achieved in a reaction system, the steps of tail end A adding and purification are reduced, the connector is added through DNA one-step reaction, and one-step library building is achieved.

The application belongs to the technical field of biology, and discloses a Yarrowia lipolytica engineering strain for efficiently producing eicosapentaenoic acid based on citric acid metabolism regulation, a method and application, wherein a chassis strain of the engineering strain is Yarrowia lipolytica Po1f, and genes of Δ9 elongase, Δ8 desaturase, Δ5 desaturase and Δ17 desaturase in a fatty acid synthesis pathway, a heterologous ATP-citrate lyase and a mitochondrial citrate carrier are overexpressed. ATP-citrate lyases and mitochondrial citrate carrier genes respectively derived from Schizochytrium sp. and Saccharomyces cerevisiae are combined to be expressed in the Yarrowia lipolytica engineering strain, so that the production of EPA and oil in the Yarrowia lipolytica engineering strain is significantly improved, and compared with a control strain, the production of oil and EPA is respectively increased by 243.35% and 161.85%.

The invention provides a carbon chain elongase as well as a coding gene and application thereof, the amino acid sequence of the carbon chain elongase is as shown in a sequence table SEQ ID NO.1, the nucleotide sequence of the gene for coding the carbon chain elongase is as shown in a sequence table SEQ ID NO.2, and the carbon chain elongase can be used as a formaldehyde lyase and has a function of efficiently catalyzing formaldehyde to synthesize 1, 3-dihydroxyacetone.

This invention discloses an O-methyltransferase gene ElCOMT33 derived from *Euphorbia lathyris* and its applications. The nucleotide sequence of this gene is shown in SEQ ID NO:1, and the encoded protein sequence is shown in SEQ ID NO:2. The protein encoded by this gene catalyzes the sequential methylation of esculetin to generate scoparone, a coumarin phytoprotective agent with disease-resistant activity. Experiments have confirmed that overexpression of ElCOMT33 in the hairy roots of *Euphorbia lathyris* significantly increases the content of scoparetin and scoparone. This invention also provides a recombinant expression vector (such as pBinPlus-GFP4) containing this gene and host cells. The ElCOMT33 gene and related materials can be used to improve the scoparone synthesis capacity in medicinal plants (such as *Euphorbia lathyris*), enhance plant disease resistance, and provide new gene resources and technical means for molecular breeding of medicinal plants.

The present invention discloses a Schizochytrium genetic engineering strain that reshapes the FAS pathway, a construction method and an application. The Schizochytrium genetic engineering strain of the present invention is obtained by heterologously expressing the C16 elongase gene E16, the C18 elongase gene E18, the Δ9 desaturase gene D9, the Δ5 desaturase gene D5, the Δ6 desaturase gene D6, and the Δ15 desaturase gene D15 in the wild-type Schizochytrium. The recombinant Schizochytrium described in the present invention can produce EPA accounting for 2.08% of the total fatty acids, and can also produce at least DHA accounting for more than 41% of the total fatty acids. In addition, the present invention claims a method for producing unsaturated fatty acids through the dual-pathway channels of PKS and FAS, which is beneficial to promoting the popularization and application of the technology for Schizochytrium to produce EPA and DHA.

This invention provides a genetically engineered *Yersinia lipolytica* strain and its applications. The genetically engineered strain is a *Yersinia lipolytica* Po1f strain with the PEX10 gene knocked out, expressing the C16 / 18 elongase gene MaELO3 from *Morchella alpina*, the elongase gene AtKCS from *Arabidopsis thaliana*, and the elongase gene CraKCS from *Brassica oleracea*. The strain also expresses the elongase gene CgKCS from *Capsella*. Further expression of the desaturase gene MaD15D from *Morchella alpina*, and overexpression of the strain's own diacylglycerol transferase gene DGA1 and its own Δ9 desaturase gene OLE1, resulted in a nervonic acid yield of 111.6 mg / L in shake-flask fermentation. By adding 1% rapeseed oil exogenously, the nervonic acid yield reached 185.1 mg / L. The *Yersinia lipolytica* genetically engineered strain constructed in this invention is simple to operate, has stable and reliable performance, and can be applied to large-scale commercial production.

The invention relates to the technical field of synthesizing a sex pheromone component precursor substance by a biological method, further relates to a method for synthesizing a moth insect sex pheromone intermediate, and particularly relates to a recombinant plasmid containing cnaphalocrocis medinalis desaturase and elongase, a recombinant cell, an expression system and a construction method and application thereof. By adopting the method provided by the invention, the sex pheromone component precursor substance can be obtained through a non-chemical synthesis method for the first time, and when the sex pheromone component precursor substance is synthesized, the method is high in specificity, free of impurities or almost free of byproducts, and environment-friendly.

The present disclosure provides a kit and its application, the kit comprising two sets of amplification primer groups, single base extension primers and processing reagents; the amplification primer groups are used for amplifying Mycobacterium tuberculosis flora identification sites and anti-tuberculosis drug pharmacogenetic polymorphism sites; each pair of primers corresponds to the upstream and downstream regions of a SNP site; the processing reagents include PCR reaction mixture, PCR enzyme mixture, SAP reaction mixture, SAP enzyme mixture, extension reaction mixture and extension enzyme mixture. By designing specific amplification primer groups and single base extension primers, the system and reagents are optimized, and the sensitivity is high. The kit combined with matrix-assisted laser desorption ionization time-of-flight mass spectrometry can complete the detection of all sites, and the detection cost is low and the efficiency is high.

The invention provides a carbon chain elongase as well as a coding gene and application thereof, the amino acid sequence of the carbon chain elongase is as shown in a sequence table SEQ ID NO.1, the nucleotide sequence of the gene for coding the carbon chain elongase is as shown in a sequence table SEQ ID NO.2, and the carbon chain elongase can be used as a formaldehyde lyase and has a function of efficiently catalyzing formaldehyde to synthesize 1, 3-dihydroxyacetone.

The invention discloses a method for increasing the yield of stearic acid in schizochytrium limacinum based on lipid droplet targeting and application, and belongs to the technical field of microbial metabolism engineering. The invention aims to solve the technical problems of non-specific positioning of single lipid droplet protein signal peptide and difficulty in efficiently producing stearic acid in schizochytrium limacinum. The invention provides a fusion protein containing a lipid droplet targeting signal peptide and palmitic acid elongase, wherein the amino acid sequence of the lipid droplet targeting signal peptide is shown as SEQ ID NO: 5, and the lipid droplet targeting signal peptide is connected to the N-terminal of the palmitic acid elongase. The fusion protein is expressed in a schizochytrium limacinum genetic engineering strain and is subjected to fermentation culture, so that the yield of stearic acid is effectively increased. The fusion protein and the method are mainly used for improving the stearic acid yield of schizochytrium limacinum, can be used for plant subcellular localization research, provide a favorable method for subcellular localization research of microalgae such as schizochytrium limacinum and have important significance.

The invention discloses an Eriocheir sinensis ELOVL7 gene and application thereof, the nucleotide sequence of the Eriocheir sinensis ELOVL7 gene is as shown in SEQ ID NO: 1, and the amino acid sequence of protein encoded by the Eriocheir sinensis ELOVL7 gene is as shown in SEQ ID NO: 2. The Eriocheir sinensis ELOVL7 gene sequence is successfully obtained through a PCR technology for the first time, the function of the ELOVL7 gene is verified through an in-vitro experiment, it is proved that the gene serves as an elongase gene to play a role in the Eriocheir sinensis fatty acid synthesis process, the function of promoting LC-PUFA synthesis is achieved, a foundation is laid for studying the LC-PUFA anabolism mechanism of the Eriocheir sinensis, and the Eriocheir sinensis ELOVL7 gene has a good application prospect. And a reference is provided for the functional research of other crustacean ELOVL7.

The invention relates to the technical field of genetic engineering algae, in particular to a method for increasing the content of triglyceride-type eicosapentaenoic acid in marine globoid cells. The method comprises the following steps: constructing an expression vector comprising a delta 5 desaturase gene, an omega 3 desaturase gene and a delta 6 elongase gene; the expression vector adopts a Ribi bidirectional promoter, one end of the Ribi bidirectional promoter is connected with a delta 6 elongase gene, a delta 5 desaturase gene and an omega 3 desaturase gene are connected through a 2A peptide sequence, a polycistron is formed in a single open reading frame, and the polycistron is connected to the other end of the Ribi bidirectional promoter; the base sequence of the delta 6 elongase gene is as shown in SEQ ID NO. 1; the base sequence of the delta 5 desaturase gene is as shown in SEQ ID NO. 2; and the omega3 desaturase gene is as shown in SEQ ID NO. 3. The expression vector is transferred into marine nannochloropsis oculata chassis cells, so that the content of triglyceride eicosapentaenoic acid in the marine nannochloropsis oculata cells is increased.

A method of inhibiting growth of cancerous cells involves contacting the cancerous cells with a very long chain fatty acid elongase 6 inhibitor (ELOVLSi). The cancerous cells are KRAS dependent. The ELOVLSi reduces a level of KRAS expressed by the cancerous cells, relative to a level thereof prior to contacting of the ELOVLSi, thereby inhibiting the growth of the cancerous cells.

Provided herein are engineered microorganisms comprising one or more genes involved in fatty acid metabolism. For example, there is provided an engineered microorganism comprising a first nucleic acid sequence encoding an elongase and a second nucleic acid sequence encoding a desaturase wherein the first and second nucleic acid sequences are operably linked to a promoter. Methods of making and using engineered microorganisms are also provided. Also provided are microbial oils comprising fatty acids, wherein the fatty acids comprise C20: 3 (n-6) (di-homo-gamma-linoleic acid) and C20: 5 (n-3) eicosapentaenoic acid (EPA). In addition, provided herein are methods for promoting the conversion of saturated fatty acids to unsaturated fatty acids by transforming a microorganism with a nucleic acid encoding a polypeptide involved in the fatty acid synthesis pathway, resulting in increased conversion of saturated fatty acids to unsaturated fatty acids.

A method for producing a microbial oil includes genetically modifying a labyrinthulid which belongs to the genus Parietichytrium or the genus Schizochytrium by disrupting and / or silencing a Δ4 desaturase gene, a C20 elongase gene, and / or a C18 elongase gene, or by being transformed with an ω3 desaturase gene; culturing the labyrinthulid; and collecting the microbial oil from the labyrinthulid. The labyrinthulid has an activity of synthesizing DHA via a PUFA-PKS pathway in an amount of not more than 1 / 100 of a total amount of DHA synthesized in the labyrinthulid or no activity of producing PUFAs via the PUFA-PKS pathway, and the labyrinthulid has an activity of producing PUFAs via an endogenous elongase-desaturase pathway.

A method of inhibiting growth of cancerous cells involves contacting the cancerous cells with a very long chain fatty acid elongase 6 inhibitor (ELOVL6i). The cancerous cells are KRAS dependent. The ELOVL6i reduces a level of KRAS expressed by the cancerous cells, relative to a level thereof prior to contacting of the ELOVL6i, thereby inhibiting the growth of the cancerous cells.

Provided herein are engineered microorganisms containing one or more genes involved in fatty acid metabolism. For example, provided are engineered microorganisms containing a first nucleic acid sequence encoding an elongase and a second nucleic acid sequence encoding a desaturase, the first and second nucleic acid sequences being operably linked to a promoter. Methods for producing and using the engineered microorganisms are also provided. Also provided are microbial oils containing fatty acids, the fatty acids including C20:3(n-6) (di-homo-γ-linoleic acid) and C20:5(n-3) eicosapentaenoic acid (EPA). Additionally, provided herein are methods for enhancing the conversion of saturated fatty acids to unsaturated fatty acids by transforming a microorganism with a nucleic acid encoding a polypeptide involved in a fatty acid synthesis pathway to result in increased conversion of saturated fatty acids to unsaturated fatty acids.

The application belongs to the technical field of biology, and discloses an exogenous cofactor enhanced Yarrowia lipolytica strain, a method and application in high-efficiency synthesis of EPA and lipids. The chassis strain of the Yarrowia lipolytica strain is Yarrowia lipolytica Po1f, and part of elongated desaturase in a fatty acid synthesis pathway, mainly including Δ9 elongase, Δ8 desaturase, Δ5 desaturase and Δ17 desaturase genes, is overexpressed in the strain. On this basis, G6PDH and malic enzyme are simultaneously overexpressed. The strain improves the EPA and lipid yield of Yarrowia lipolytica. The application is suitable for constructing a gene engineering strain with high EPA and lipid yield, and provides a basis for Yarrowia lipolytica to produce EPA-rich oil, so as to solve the problems of unsustainability and high cost caused by obtaining EPA from fish.

This invention belongs to the fields of genetic engineering and fermentation engineering, and discloses a method for constructing and applying a Schizochytrium strain that produces high DHA content. The engineered Schizochytrium strain uses Schizochytrium sp. HX308 as the starting strain. Through metabolic engineering, a complete elongation-desaturation pathway is established in Schizochytrium (overexpression of C16 / 18 elongase, Δ-9 elongase gene, Δ-5 desaturase gene, Δ-17 desaturase gene, Δ-5 elongase gene, and Δ-4 desaturase gene), which works synergistically with the PKS pathway to produce DHA. The peroxisome synthesis factor PEX10 is knocked out to obtain the recombinant engineered strain SCD3. In fed-batch fermentation, the final DHA content of the engineered Schizochytrium strain SCD3 is the highest to date, reaching 70.1% of the total fatty acid content. The oil and DHA yields are also significantly increased, reaching 67.9 g / L and 47.6 g / L, respectively. The DHA yield is 1.7 times that of the wild type. This engineered strain has good application prospects.