882results about "Acyltransferases" patented technology

The invention relates to a recombinant pichia pastoris strain capable of producing astaxanthin at high yield as well as a construction method and application of the recombinant pichia pastoris strain. The recombinant pichia pastoris strain is obtained by expressing ATP (adenosine triphosphate) citrate lyase (ACL) and acetyl-CoA synthase (ACS) or phosphoketolase (PK) and phosphotransacetylase (PTA), choline kinase (CK), inositol polyphosphate kinase (IPK) and vitreoscilla hemoglobin (VHB) in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified strain PP-LC2. The astaxanthin production performance of the recombinant strain is verified on the basis of comparison of precursor supply pathways, IUP pathway construction and improvement of the oxygen supply capacity of the engineering strain, and the astaxanthin production capacity of the pichia pastoris is further improved. The construction method of the recombinant pichia pastoris is simple, the synthesis of astaxanthin can be better promoted, the engineering strain can efficiently synthesize the astaxanthin by utilizing methanol through amplification fermentation of a 5L fermentation tank, and industrial production is facilitated.

The invention relates to an L-valine production strain as well as a construction method and application thereof, the strain is obtained by transformation through a metabolic synthesis path of L-valine, and the construction method comprises the following steps: by taking Cornebacterium glutamicum ATCC 13032 (C.glutamicum) as an original strain, carrying out editing transformation such as gene knockout, gene overexpression, gene heterologous expression, gene double-copy expression and the like on a genome of the original strain by utilizing a metabolic engineering means. The L-valine production strain disclosed by the invention can be used for synthesizing L-valine from the beginning by taking glucose as a carbon source, has the advantages of high production rate, short fermentation period, high strain stability and low cost, and lays a foundation for realizing large-scale production of L-valine. According to the fermentation tank fermentation method, the L-valine is produced in a mechanical stirring type fermentation tank by using the L-valine production strain Va1-09, 86.4 g / L of L-valine is produced by fermenting for 36 hours, and the method has a good industrial prospect.

The present disclosure relates to isolated nucleic acid molecules comprising at least two double-stranded inhibitory ribonucleic acid (RNA) molecules adapted to silence the same or different genes by RNA interference to enhance silence, thereby modulating gene expression.

The present disclosure relates to compositions and methods of increasing levels of fetal hemoglobin (HbF) in cells. The present disclosure further relates to methods for treating patients suffering from blood cell diseases, including those associated with reduced amounts of functional adult hemoglobin (HbA), such as sickle cell disease and β-thalassemias

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 relates to recombinant saccharomyces cerevisiae for de novo synthesis of mevalonic acid as well as a construction method and application of the recombinant saccharomyces cerevisiae, and belongs to the technical field of genetically engineered bacteria. The technical problem to be solved by the invention is to provide the recombinant saccharomyces cerevisiae for de novo synthesis of mevalonic acid. According to the recombinant saccharomyces cerevisiae, saccharomyces cerevisiae is used as a chassis strain, and ACL, CTP1, RtME and MDH3 are over-expressed. A series of recombinant saccharomyces cerevisiae strains with high yield of mevalonic acid are constructed through a gene recombination method, the strains can be fermented in a conventional culture medium to prepare mevalonic acid, the fermentation process is simple, the cost is low, the yield is high, and the maximum yield of mevalonic acid can reach 5692.45 mg / L. According to the invention, the recombinant saccharomyces cerevisiae strain is used as a chassis strain, and recombinant genetically engineered bacteria for producing the santalene, the santalol and the patchouli alcohol are respectively obtained through gene recombination, so that the yields of the santalene, the santalol and the patchouli alcohol produced by fermentation are improved.

The invention provides a method for producing borneol, the method comprising culturing the transgenic microorganism in a culture medium, and the microorganism comprises or expresses borneol diphosphate synthetase and phosphatase.

The present disclosure provides guided microbial remodeling (GMR) methods for the rational improvement of plant-associated microbes to perform plant-beneficial functions. The GMR methods described herein allow for non-intergeneric genetic optimization of key regulatory networks within the microbes, which improve plant-beneficial functions over wild-type microbes but don't have the risks associated with transgenic approaches (e.g., unpredictable gene function, public and regulatory concerns, etc.). The present disclosure also provides remodeled microbes and compositions thereof. The utilization of remodeled microbes and compositions thereof will enable farmers to realize more productive and predictable crop yields without the nutrient degradation, leaching, or toxic runoff associated with traditional synthetically derived fertilizers.

The invention relates to a diacylglycerol acyltransferase capable of synthesizing triglyceride as well as a coding gene and application of the diacylglycerol acyltransferase, and belongs to the field of enzymology, the nucleotide sequence of the gene for coding the diacylglycerol acyltransferase is as shown in SEQ ID NO.1, and the amino acid sequence of the diacylglycerol acyltransferase is as shown in SEQ ID NO.2. The invention also provides a recombinant plasmid and a recombinant engineering bacterium containing the gene of the enzyme. Diglyceride and an acyl donor are used as substrates, and triglyceride is catalytically synthesized through an acyl transfer reaction by using the enzyme. The invention provides an efficient tool for realizing enzymatic biosynthesis and large-scale production of natural triglyceride.

This invention provides a genetically engineered bacterial strain, its preparation method, and its application in the synthesis of perillaldehyde. The genetically engineered bacterial strain includes the CcLPPS gene derived from *Cistus creticus*; the strain is selected from *Yarrowia lipolytica*. This invention constructs a stable, highly efficient de novo perillaldehyde synthesis strain. The entire fermentation process requires no addition of any organic reagents or intermediate substances, significantly reducing production costs and simplifying subsequent separation and purification steps. The engineered *Yarrowia lipolytica* strain provided by this invention exhibits high production intensity and a simple fermentation process, showing promising application prospects and laying the foundation for the microbial fermentation production of perillaldehyde.

The present invention belongs to the field of biotechnology. Disclosed is the use of an NAT protein of Oryza sativa and an encoding gene thereof in the regulation of plant disease resistance. In the research on the immune response of Oryza sativa, one gene NAT is identified, the expression of which is induced by Magnaporthe oryzae, is obtained by means of identification. Experimental verification shows that overexpression of the NAT gene in Oryza sativa can significantly improve the disease resistance of Oryza sativa. It is indicated that the NAT gene positively regulates the disease resistance of plants, and thus the NAT gene can be used as a target gene for molecular breeding to improve the disease resistance of Oryza sativa or other plants, thereby laying a theoretical basis for research into plant disease resistance.

The invention relates to recombinant escherichia coli for de novo synthesis of piceatannol as well as a construction method and application thereof, and belongs to the field of synthetic biology and metabolic engineering. According to the invention, an L-tyrosine production strain is used as an original strain, five key genes required for synthesizing piceatannol are combined and connected to a plasmid vector, and a recombinant plasmid is introduced into the original strain, so that an engineering strain for heterologous synthesis of piceatannol is successfully constructed. According to the invention, the yield of piceatannol is further obviously improved through copy number optimization.

A recombinant yeast cell functionally expressing: a) a nucleic acid sequence encoding a protein comprising phospho-ketolase (PKL) activity (EC 4.1.2.9 or EC 4.1.2.22) and / or a nucleic acid sequence encoding a protein having phosphotransacetylase (PTA) activity (EC 2.3.1.8) and / or a nucleic acid sequence encoding a protein having acetate kinase (ACK) activity (EC 2.7.2.12); and / or) a nucleic acid sequence encoding a protein having transketolase activity (EC 2.2.1.1), wherein the expression of the nucleic acid sequence encoding the protein having transketolase activity is under control of a promoter (the “TKL promoter”), which TKL promoter has an anaerobic / aerobic expression ratio for the transketolase of (2) or more.

The invention provides a resveratrol production strain and a construction method and application thereof, according to the strain, tyrAfbr, tyrB, aroE, pntAB, ompF, aroGfbr and accA genes are up-regulated, a carbon storage regulation factor csrA gene is knocked out, the transcriptional level of a fabF gene is down-regulated through a BBaJ23113 promoter, a TAL gene from Rhodotorula glutinis, a 4CL gene from Arabidopsis thaliana and an STS gene from Vitis vinifera are heterologously expressed on a genome, and the strain carries a high-copy plasmid PET-28a-WAL at the same time; the strain takes glucose as a carbon source, resveratrol is efficiently and stably synthesized from the beginning by adopting a fermentation method, the production cost is low, and efficient production of resveratrol is realized.

This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. This disclosure describes a cell metabolically engineered for production of a mixture of at least three different oligosaccharides. Furthermore, this disclosure provides a method for the production of a mixture of at least three different oligosaccharides by a cell as well as the purification of at least one of the oligosaccharides from the cultivation.

The invention provides a strain for producing L-isoleucine as well as a construction method and application thereof. The strain is obtained by modifying a chassis strain escherichia coli XX12 by utilizing a metabolic engineering modification method, ldhA, adhE and pflB genes are deleted, the transcriptional levels of aspC, pykF, pntAB, ppK, ppc, ilvAfbr, ilvIHfbr and ygaZH genes are up-regulated, the transcriptional level of a leuA gene is down-regulated, a bcd gene derived from B.subtilis 168 and ppnK and cysK genes derived from Cornebacterium glutamicum ATCC 13032 are heterologously expressed, and the strain has the advantages that the strain can be used for producing L-isoleucine; the method has the advantages of no need of adding resistant substances, good L-isoleucine synthesis capability, short fermentation period by using glucose as a carbon source, and good economic benefit and industrial application value.

Described is an antibody-drug conjugate (ADC) having the formula A-L, wherein A is an antibody or an antibody fragment and wherein L is a linker, said linker comprising: as a first payload a topoisomerase I inhibitor which is cell-permeable, e.g., a camptothecin cytotoxic molecule which is cell-permeable; as a second payload a topoisomerase I inhibitor which is not cell-permeable, e..g., a camptothecin cytotoxic molecule which is not cell-permeable; and as a as a third payload a toxin or a cytotoxin, e.g., an auristatin such asMMAE (Monomethyl auristatin E). Moreover, described is a pharmaceutical composition comprising said ADC and at least one pharmaceutically acceptable ingredient. Further, described method of treating a patient suffering from, being at risk of developing, and / or being diagnosed for a neoplastic disease.

The disclosure provides methods for increasing saturated, monounsaturated, and poly unsaturated fatty acid production using the PUFA gene cluster plasmid and the gene regulator pfaR.

An application of rice NAT protein and a coding gene thereof in regulation and control of plant disease resistance, a method for regulating and controlling rice resistance to rice blast and a method for cultivating transgenic rice with high resistance to rice blast are provided. In the research of rice immune response, a gene NAT of which the translation is induced by M. oryzae is identified and obtained.

The present disclosure provides a method of treating estrogen receptor-positive (ER+) breast cancer, comprising the combined administration of therapeutically effective amounts of a KAT6A / B inhibitor and a Menin inhibitor to a patient in need thereof.

The invention belongs to the technical field of genetic engineering, and particularly relates to a genetically engineered bacterium for producing N-acetylneuraminic acid as well as construction and application of the genetically engineered bacterium. Accumulation of acetic acid is controlled by combining and applying different strategies: rational transformation is performed on an escherichia coli metabolic pathway, and acetic acid synthesis pathway related genes poxB, ackA and pta are knocked out to reduce acetic acid synthesis; the glyoxylic acid cycle is activated by knocking out the coding gene iclR of the glyoxylic acid cycle transcriptional regulation factor, and acetic acid overflow is reduced; the fermentation temperature is adjusted to control overall metabolism of cells to reduce acetic acid accumulation. Through comprehensive application of the strategy, the Neu5Ac production method capable of effectively controlling acetic acid accumulation is obtained, and the method has a good industrial application prospect.

The invention relates to the technical field of genetic engineering, in particular to a gene for biosynthesis of (Z)-13-octadecenoic acid and application of the gene. The invention discloses a gene for biosynthesis of (Z) 13 octadecenoic acid. An amino acid sequence coded by the gene is as shown in SEQ ID NO: 1. The invention finds that the (Z)-13-octadecenoic acid can be synthesized by jointly expressing the protein with the amino acid sequence shown as SEQ ID NO: 1, hexadecanoic acid specific thioesterase and Z11 desaturase in plants. The process of synthesizing the (Z)-13-octadecenoic acid by using plants is green and pollution-free, the purification is simple and convenient, the environmental pollution degree is low, and the extraction purity is high.

The invention belongs to the technical field of gene engineering, and particularly relates to application of a GhHAT18 gene in regulation and control of drought stress resistance of plants, the nucleotide sequence of the GhHAT18 gene is shown as SEQ ID NO.1. The invention discloses that the cotton histone acetylase gene GhHAT18 can be used for regulating and controlling the drought resistance of the plants for the first time, and experiments show that compared with a wild type plant, after the expression of the GhHAT18 gene is inhibited, the GhHAT18 gene can be used for regulating and controlling the drought stress resistance of the plants. The antioxidant enzyme activity and the drought resistance of plants can be obviously reduced; and the transgenic plant over-expressed with the GhHAT18 shows stronger and higher antioxidant enzyme activity and drought resistance under the drought stress. Transcriptome analysis shows that the GhHAT18 gene affects the expression of drought-resistant related genes through a secondary metabolite synthesis pathway and an MAPK signal transduction pathway, and the adaptive capacity of plants to drought stress is further regulated and controlled.

The invention belongs to the technical field of animal model construction, and particularly relates to a non-alcoholic fatty liver disease mouse model based on humanized PNPLA3 I148M transgene as well as a construction method and application thereof, the construction method comprises the following steps: step i: designing CDS of a human gene and a key 3 'UTR element for patent medicine of a small nucleic acid drug in gene plasmid DNA of the constructed model; step ii, using Luc2 or iRFP to report gene expression in the gene plasmid DNA for constructing the model; and (iii) a disease model of the fatty liver disease induced by combining plasmid DNA transient transfer with high-fat diet. The mouse model constructed by the invention has the advantages that a specific exogenous gene expression signal of the liver is detected in real time through small animal living imaging, a human PNPLA3 protein is detected and overexpressed through serum ELISA, PNPLA3 I148M mRNA is detected and overexpressed through liver RT-qPCR, the phenotype of the non-alcoholic fatty liver disease of the liver is determined through 15-week-old pathology, and the whole process of modeling for 8 weeks does not need to be operated in a biosafety secondary laboratory.

The invention discloses a recombinant escherichia coli strain, a preparation method and application thereof, and a synthesis method of oryzanol A, and relates to the technical field of bioengineering. The recombinant escherichia coli strain takes E.coli BL21 (DE3) as a chassis cell, and is modified as follows: (a) knockout of a gene pfkA; or, the gene pfkA is knocked out, and an untranslated region of the 5'end of the gltA gene is mutated from the sequence as shown in SEQ ID NO: 1 to the sequence as shown in SEQ ID NO: 2; and (b) inserting exogenous genes agmA, agmB, agmC, agmE and agmF, so as to obtain the gene. The recombinant Escherichia coli strain is high in growth speed and short in fermentation period (within 72 hours), a culture medium which is cheap and easy to obtain can be adopted, and compared with an existing Escherichia coli system, the fermentation yield of oryzanol A is obviously increased.

The invention discloses a recombinant bacillus paralicheniformis and an application of the recombinant bacillus paralicheniformis in production of betaine. According to the invention, bacillus paralicheniformis Bp1 with heavy metal cadmium resistance is used as a chassis strain, and an de novo synthesis pathway which takes glucose as a substrate and synthesizes glycine betaine through glycolysis, glyoxylic acid pathway and glycine methylation pathway in sequence is established in the bacillus paralicheniformis, so that the survival rate of the glycine betaine in cadmium-polluted soil is increased; the effects of the microbial soil treating agent and the microbial fertilizer are improved, so that the growth of the plant-black nightshade with an enrichment effect on cadmium and the absorption of the black nightshade on the cadmium in the polluted soil are promoted.

Provided herein are methods for coating a microparticle, cell, or therapeutic agent with a biomimetic zona pellucida (BZP). Also provided herein are BZP-coated cells prepared using the enclosed methods. The methods for using the BZP-coated microparticle to treat diabetes are also provided.

The invention provides a high-yield O-succinyl-L-homoserine engineering bacterium as well as a construction method and application of the high-yield O-succinyl-L-homoserine engineering bacterium. According to the invention, O-succinyl-L-homoserine genetically engineered bacteria with high yield of O-succinyl-L-homoserine are constructed by introducing O-succinyl-homoserine transferase from Vibrio cholera into a chassis strain in an exogenous manner and obtaining a gene mutant metAVc (T242A) of the O-succinyl-L-homoserine transferase, and further increasing the expression of the O-succinyl-L-homoserine transferase by adopting a Trc promoter. The engineering strain obtained through a systematic metabolic engineering modification strategy can realize effective accumulation of OSH, the shake flask yield of OSH reaches 18.8 g / L, the fed-batch fermentation yield of a 5L fermentation tank reaches 105.4 g / L, the sugar-acid conversion rate reaches 52.6%, and a foundation is laid for subsequent construction of high-yield OSH engineering bacteria.

This invention specifically relates to a de novo 7-DHC-synthesizing *Saccharomyces cerevisiae* genetically engineered strain, its construction method, and its applications, belonging to the field of biotechnology. Using *Saccharomyces cerevisiae* engineered strain SC9 as the starting strain, this invention utilizes multiple copies of tHMG1, DHCR24, and ERG1 via the TY transposon, multiple copies of ERG2, POS5, ERG7, ERG11, IDI1, and ERG3 via the TY transposon, and single copies of POS5, ERG3, and DHCR24 in the genome to increase the intracellular content of reducing coenzyme II and enhance the main pathway of 7-DHC synthesis. Single copies of ACL, ACS1, ERG10, and ERG19 in the genome enhance the utilization of ethanol in *Saccharomyces cerevisiae* and dynamically regulate the synthesis of 7-DHC. The *Saccharomyces cerevisiae* genetically engineered strain constructed by this invention exhibits high product accumulation, laying the foundation for replacing traditional multi-step chemical synthesis routes and realizing industrial production.

The invention discloses a genetically engineered bacterium for high yield of L-alanine as well as a construction method and application of the genetically engineered bacterium, a dominant strain is obtained by knocking out an alanine racemase gene dadX and integrating coding sequences of alanine dehydrogenase from different sources, and the shake flask yield is 10.8 g / L; the activity of a corresponding mutant M18L / E75A / A78S / I266V is 4.3 times that of a wild type; 6 by-product related genes are knocked out, meanwhile, a multi-copy strain integrated with alanine dehydrogenase is constructed, and the yield of E.coli A-LB6 in a 5L fermentation tank is increased to 133.3 g / L. Through a dynamic regulation and control strategy, an oxygen-sensitive promoter Pomp * is adopted to replace a phosphoenolpyruvate carboxylase promoter, so that succinic acid byproducts are reduced by 85.8%, and the yield reaches 143.5 g / L after 48 hours of aerobic and anaerobic two-stage fermentation. And finally, by regulating and controlling the ATP synthetase gene cluster atp promoter, the yield of L-alanine in a 50 L fermentation tank reaches 151.3 g / L, and the saccharic acid conversion rate reaches 95.2%. The combination of polygene knockout and dynamic metabolism regulation effectively enhances the biosynthesis ability of alanine, and provides an efficient engineering strain for industrial production.