1350results about "Acyltransferases" patented technology

The present disclosure concerns a fusion polypeptide for making acetyl-phosphate from glucose, a recombinant yeast host cell expressing the fusion polypeptide as well as methods of using the fusion polypeptide. The fusion polypeptide comprising (i) a glucose-6-phosphate isomerase moiety and (ii) a phosphoketolase moiety.

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 the technical field of dye synthesis, and particularly discloses a genetically engineered bacterium for synthesizing N-acetyl blue as well as a construction method and application of the genetically engineered bacterium. The supply capacity of precursor substances glutamic acid and glutamine of the genetically engineered bacterium is high, the fermentation process is simple, the production intensity of the precursor substances glutamine and N-acetylglutamine is improved by cutting off a consumption path, relieving negative feedback inhibition and enhancing the expression level of key genes through the genetically engineered bacterium, and finally the yield of N-acetylblue reaches 41.2 g / L.

Provided herein, inter alia, is an engineered non-plant cell that produces a hyoscyamine alkaloid product, a precursor of a hyoscyamine alkaloid product, or a derivative of a hyoscyamine alkaloid product. Also described is a method for producing a scopolamine alkaloid, a precursor of a scopolamine alkaloid product, or a derivative of a scopolamine alkaloid product using a cell.

The invention discloses an emodin anthraquinone high-yield engineering strain. According to the invention, mdpG and mdpF genes of aspergillus nidulans are exogenous introduced into a chassis strain aspergillus nidulans LO8030, so that the genetically engineered bacterium for producing emodin is obtained. Furthermore, the g1234 gene is knocked out from the genetically engineered bacterium for producing the emodin, so that the yield of the emodin can be obviously improved. The decarboxylase gene is further introduced in an exogenous manner, so that the yield of the emodin can be further remarkably improved. And on the basis, exogenous introduction of mdpK, mdpC and mdpB genes is carried out, such that the chrysophanol-producing genetically engineered bacterium is obtained. Or the rugG gene from talaromyces sp.YE3016 is introduced in an exogenous manner, so that the genetically engineered bacterium for producing the muscardine hydrochloride is obtained.

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 invention relates to a method for increasing the yield of L-malic acid produced by fermentation by adding ethylene glycol, which comprises the following steps: constructing genetic engineering escherichia coli, and introducing ethylene glycol metabolism genes (overexpression) including glycerol dehydrogenase, lactaldehyde reductase, lactaldehyde dehydrogenase and the like; and overexpression of other genes involving malic acid production, such as malic acid synthase, phosphoenolpyruvate carboxylase and malic acid export protein. The gene engineering strain is an E.coli BW25113 strain of which ldhA, pta, poxB, adhE, pflB, mgsA, frdA, maeA, maeB, yqhD, ycdW and iclR genes are substantially inactivated, and the deleted genes are beneficial to reducing the generation of metabolic byproducts and increasing the accumulation of malic acid. The genetic engineering strain is subjected to fermentation culture by taking glucose and ethylene glycol as carbon sources, and L-malic acid is separated from a culture after culture is finished.

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 present invention provides a genetically engineered bacterium for synthesizing N-acetylglucosamine, a preparation method thereof, and an application thereof. The genetically engineered bacterium uses Escherichia coli as an initial bacterium, and related genes for the catabolism of N-acetylglucosamine are knocked out on the genome of the initial bacterium, and related genes for the metabolic synthesis of N-acetylglucosamine are overexpressed, so as to improve the production level of N-acetylglucosamine. Among them, the related genes for the catabolism of N-acetylglucosamine include any one or a combination of at least two of nagB, nagA, manX, nagE, nagK, or ptsG, and the related genes for the metabolic synthesis of N-acetylglucosamine include any one or a combination of at least two of glmS, gna1, glnA, yqaB, icaC, galP, or glk.

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 invention discloses recombinant streptomyces mobaraensis as well as a construction method and application thereof, and belongs to the technical field of bioengineering. According to the invention, smY2019 is used as an original strain, and a CRISPR / cas9 technology is utilized to construct five chassis cells of a GIs gene cluster; on the basis of smY2019-deltaG5, two kinds of chassis cells in which endogenous CRISPR / Cas system genes are knocked out are constructed. And on the basis of the smY2019-deltaG5-C2, further constructing a chassis cell smY2019-deltaG5-C3 of which a non-homologous end connection (NHEJ) repair pathway gene is knocked out. The TGase secretory enzyme activity of the chassis cell smY2019-deltaG5-C3 is improved by 37.3% compared with that of the original strain smY2019, and meanwhile, the transformation efficiency, the growth speed, the protein synthesis capability and the energy metabolism level of the chassis cell are obviously improved compared with that of the original strain smY2019.

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 belongs to the technical field of strain metabolism modification, and particularly relates to a streptomyces avermitilis strong promoter and application. The nucleotide sequence of the streptomyces avermitilis strong promoter provided by the invention is as shown in SEQ ID NO. 4. The strong promoter can express and regulate a key gene, and the mRNA relative expression level of the gene driven by the strong promoter can still be kept stable in the 48-96-hour growth stage of the streptomyces avermitilis, which indicates that the strong promoter can keep high activity in the 48-96-hour growth stage of the streptomyces avermitilis, so that the target gene can be stably and highly expressed. Therefore, the strong promoter can be used for constructing recombinant plasmids for expressing key genes for synthesizing target natural products and streptomyces avermitilis recombinant bacteria, and the constructed streptomyces avermitilis recombinant bacteria can efficiently produce the target natural products.

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.

The present invention belongs to the field of genetic engineering technology and discloses a plasmid-free genetically engineered bacterium that can efficiently synthesize γ-aminobutyric acid from scratch using a cheap carbon source as a substrate, a method and an application thereof. The engineered bacterium is based on wild-type E. coli MG1655, overexpresses the T7 RNA polymerase gene; deletes the genes gabT and puuE; overexpresses gadC and gad bm , gdh, gltA, pyc, and ppc genes; and a growth-coupled promoter was used to dynamically regulate the expression of the sucA and argA genes in the GABA production pathway in Escherichia coli. All genetic manipulations in this engineered bacterium were performed within the genome, leaving no plasmid residues and requiring no antibiotics or inducers. The resulting strain exhibited stable production performance and a simple fermentation process. Using the engineered bacterium under a staged pH control process for 38 hours, the yield of γ-aminobutyric acid reached 35.4 g / L, demonstrating promising industrial application potential.

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.

A genome-editing complex for modifying a target polynucleotide comprising a recombinant β helical protein linked to either one or more molecules of a genome-editing system or a plasmid encoding for one or more molecules of a genome-editing system, wherein the β helical protein length is in the range of from 5 nm to 25 nm, and width is in the range of from 1 nm to 5 nm.

The present invention is in the field of genetically modified enzyme and microorganism comprising such a modified enzyme for the production of raspberry ketone or zingerone. The modified enzyme is a polyketide synthase (PKS) issued or derived from a wild type PKS. The modified PKS has the capability to produce vanillylidene acetone from feruloyl-CoA and / or 4-hydroxybenzalacetone from 4-coumaroyl-CoA in a more effective way as compared to wild type BAS or wild type PmPKS, in particular in recombinant bacteria strains.

The present invention relates to the field of biotechnology, and particularly to the application of recombinant Saccharomyces cerevisiae in the production of homopterocarpin using p-coumaric acid. The present invention provides an engineered Saccharomyces cerevisiae capable of producing homopterocarpin using p-coumaric acid. Through strategies such as artificial pathway construction, pathway optimization, rational design of enzymes, and fermentation process optimization, the present invention obtains an engineered Saccharomyces cerevisiae capable of producing homopterocarpin using p-coumaric acid. Experiments show that its homopterocarpin yield is 1.005 mmol / l and the conversion rate is 65.8%. In large-scale production in a 5L fermenter, the strain of the present invention can utilize 15.2 mmol / l of p-coumaric acid to produce 3.2 mmol / l of homopterocarpin, and the conversion rate is 21.1%.

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.