80 results about "Acyl carrier protein" patented technology

The invention relates to crop plants comprising novel seed lipid compositions. Provided are both wild type and mutant nucleic acid molecules encoding Brassica fatty acyl-acyl carrier protein (ACP) thioesterase B proteins (FATB) and the proteins as such. Also provided are Brassica plants, tissue and seeds comprising at least three mutant fatB alleles in their genome, whereby the seed oil fatty acid composition or profile is significantly altered.

The present invention relates to compounds of the Formula (I), wherein X1 and X2 independently represent O, NH or S; R1 and R2 are independently selected from the group consisting of a C1-C30 hydrocarbyl group, an amino acid bonded via an amide bond or a peptide bonded via an amide bond each having up to 200 amino acids, the conjugated residue X1 or X2 in this case being NH, and hydrogen, both radicals R1 and R2 preferably not being H; and R3 is a residue selected from group consisting of —S—R6, wherein R6 is a C1-C30 hydrocarbyl group, at least one of R1 and R2 not being H when X1 and X2 are oxygen, —S—CH2—CH(NH2)(COOH) (cysteine-S-yl), a homologue or derivative (e.g. N-acetyl cysteine-S-yl) thereof, a peptide having up to 200 amino acids which contains at least one amino acid radical with a thiol group, preferably a cysteine radical, and is bonded via the thio sulfur, preferably via the cysteine sulfur (peptide-S-yl), coenzyme A which is bonded via a thiol group or fragments thereof, acyl carrier protein bonded via a thiol group, and dihydrolipoic acid bonded via a thiol group; and pharmaceutically acceptable salts thereof. The present invention also relates to the use of these compounds for preparing a drug, drugs containing the same and their use for the therapy of diseases such as autoimmune disease, NF-kappaB mediated diseases, psoriasis, psoriatic arthritis, neurodermitis, enteris regionalis Crohn, cardiac insufficiency, chronic obstructive pulmonary diseases and asthma and in transplantation medicine.

The present invention relates to compounds of the formula (I) wherein X1 and X2 independently represent O, NH or S, R1 and R2 are independently selected from the group consisting of a C1-C30 hydrocarbyl group, an amino acid bonded via an amide bond or a peptide bonded via an amide bond each having up to 200 amino acids, the conjugated residue X1 or X2 in this case being NH, and hydrogen, both radicals R1 and R2 preferably not being H, wherein R3 is a residue selected from group consisting of —S—R6, wherein R6 is a C1-C30 hydrocarbyl group, at least one of R1 and R2 not being H when X1 and X2 are oxygen, —S—CH2—CH(NH2)(COOH) (cysteine-S-yl), a homologue or derivative (e.g. N-acetyl cysteine-S-yl) thereof, a peptide having up to 200 amino acids which contains at least one amino acid radical with a thiol group, preferably a cysteine radical, and is bonded via the thio sulfur, preferably via the cysteine sulfur (peptide-S-yl), coenzyme A which is bonded via a thiol group or fragments thereof, acyl carrier protein bonded via a thiol group, and dihydrolipoic acid bonded via a thiol group, and pharmaceutically acceptable salts thereof. The present invention also relates to the use of these compounds for preparing a drug and drugs containing the same.

Process for conjugation of bacterial saccharides including Streptococcus pneumoniae and Haemophilus influenzae saccharides by reductive amination are provided herein.

The invention relates to a production method for synthesizing C8:0 / C10:0 / C12:0 / C14:0 medium-chain fatty acid and ethyl ester thereof by using engineering Escherichia coli. The production method comprises the following steps of: cloning thioesterase genes with different specificities chFatB / ucFatB / ccFatB in cuphea / laurel / camphorwood into an expression vector pETDuet-1 to obtain a corresponding recombinant plasmid; transferring the recombinant plasmid into Escherichia coli BL21, performing isopropyl-beta-d-thiogalactoside (IPDT) induction expression, and specifically hydrolyzing acyl carrier protein (ACP) with specific chain length by using the expressed thioesterase to obtain medium-chain fatty acid; cloning acyltransferase gene (atfA) in acinetobacter into expression vectors pETDuet-chFatB / pETDuet-ucFatB / pETDuet-ccFatB to obtain corresponding recombinant plasmids; and transferring the recombinant plasmids into the Escherichia coli BL21, performing IPTG induction expression, performing acyl transfer reaction on exogenously fed ethanol and acyl-CoA formed by the medium-chain fatty acid intracellularly by using the co-expressed acyltransferase to obtain fatty acid ethyl ester.

The invention relates to a monoclonal antibody of hepatitis B virus X protein (HBx), which is characterized by having specificity reaction to N-end epitope or C-end epitope of HBx, but having no reaction to keyhole limpet hemocyanin (KLH) of carrier protein and other expressed proteins of hepatitis B virus (HBV). The preparation method of the monoclonal antibody comprises the steps of: fusing N-end and C-end antigen epitope polypeptides of HBx respectively with mice immunized with KLH in crosslinking way, and myeloma cells and screening to obtain the monoclonal antibody. The monoclonal antibody can be used for various immunodetection reagents and directed therapeutic drugs of HBx protein or HBx antibody and applied to diagnosis and treatment of diseases such as hepatitis b virus (HBV) infection, hepatocellular carcinoma (HCC) and the like.

The present invention relates to a vector for the expression of recombinant proteins, antigens, pathogen-like particles and immunogenic complexes, said vector (pMRKA vector) being produced by modifying the plasmids containing the gene sequence of the T7 promoter of E. coli, this modification being mainly characterized by the substitution of the ampicillin-resistance gene by the kanamycin-resistance gene, and by the insertion of the par sequence (partition sequence which determines the efficient segregation of the plasmids in daughter cells during cell division). Also provided are expression vectors based on the pMRKA plasmid, which additionally comprise at least one of the gene sequences of the exosome of P. abyssi, which vectors are designated pMRKA-EXO, pMRKA-RING and pSUMAC. The invention also provides the vectors additionally comprising gene sequences with immunomodulatory or immunoregulatory activity, preferably the pMRKA-Z-Z-EXO and pMRKA-Z-Z-RING vectors. Other aspects of the invention include the method for producing said expression vectors and the use of the obtained vectors.

Process for conjugation of bacterial saccharides including Streptococcus pneumoniae and Haemophilus influenzae saccharides by reductive amination are provided herein.

The invention discloses a bacterial polysaccharide O-glycosylation modified recombinant cholera toxin B subunit fusion protein and an application thereof. The present invention provides a conjugate of polysaccharide and protein which is coupled by the bacterial polysaccharide and recombinant cholera toxin B subunit fusion protein. The bacterial polysaccharide is connected with the glycosylation sites of the recombinant cholera toxin B subunit fusion protein in the form of O-glucosidic bond. The experiment indicates that the preparation of the bacterial polysaccharide protein conjugate vaccine from O-glycosylation modified recombinant cholera toxin B subunit fusion protein can enhance the ability of inducing animals to generate anti-polysaccharide antibodies, avoid miscellaneous problems of pathogen culture, enhance the vaccine homogeneity and production efficiency, and reduce the vaccine preparation cost, therefore possessing wide application prospect.

The invention provides a synthetic method of urethane artificial antigen. The urethane artificial antigen is prepared by the following steps of: 1, preparation of beta-alanine and ethyl chloroformate; 2, synthesis of hapten; 3, purification of hapten; 4, preparation of 1-(3-dimethylamino propyl)-3-ethyl carbodiimide solution; 5, synthesis of artificial antigen; 6, dialysis and detection of artificial antigen; and 7, calculation of quantity of different used carrier proteins. According to the synthetic method, the shielding effect of the protein to the hapten obtained by chemical synthesis is low, therefore, high-titer polyclonal antibody can be generated in an immune animal, and the molecular weight can be detected during synthesizing the artificial antigen, and the coupling ratio can be calculated, and the quantity of used raw materials for synthesizing is precise.

This disclosure describes genetically modified photosynthetic microorganisms, e.g., Cyanobacteria, that overexpress an acyl-acyl carrier protein reductase (acyl-ACP reductase). These microorganisms may optionally overexpress one or more fatty acid synthesis proteins such as ACP and ACCase, and / or one or more polypeptides associated with glycogen breakdown. Also included are photosynthetic microorganisms comprising mutations or deletions in a glycogen biosynthesis or storage pathway, which accumulate a reduced amount of glycogen under reduced nitrogen conditions as compared to a wild type photosynthetic microorganism. The modified photosynthetic microorganisms provided herein are capable of producing increased amounts of lipids such as fatty acids or wax esters and / or synthesizing triglycerides.

Disclosed herein are antibacterial compounds that inhibit fabl, a NADH-dependent enoyl [acyl carrier protein] reductase enzyme in the fatty acid biosynthesis pathway. The compounds are represented by structural formulas Ia and Ib: R1 and R2 are independently monocyclic aryl or heteroaryl groups, wherein the groups represented by R1 and R2 are optionally substituted with one or more acyclic substituents; R3 is —H or an optionally substituted C1-C8 aliphatic, C3-C8 cycloaliphatic, aryl, or heteroaryl group. X1 is a bond or a C1-C3 alkylene chain that is optionally substituted with a C1-C4 alkyl or an acidic group. X2 is an aryl, heteroaryl or C3-C8 cycloaliphatic ring, wherein the group represented by X2 is optionally substituted with triazole, tetrazole, and / or one or more acyclic substituents.

The invention discloses a construction method and application of schizochytrium limacinum engineering bacteria for KS (Beta-ketoacyl Synthase) gene knockout. The construction method comprises the following steps: (1) using a genome of schizochytrium limacinum as a DNA (Deoxyribonucleic Acid) template, and respectively carrying out PCR (Polymerase Chain Reaction) amplification on an upstream fragment UKS and a downstream fragment DKS of a KS gene by using an upstream primer pair and a downstream primer pair; (2) connecting the upstream fragment UKS and the downstream fragment DKS with a knockout carrier, and constructing recombinant knockout plasmids; (3) electrically converting the recombinant knockout plasmids into the schizochytrium limacinum, screening by using a resistant plate, and verifying through a PCR resistant gene sequence, thus obtaining a converter, i.e., the schizochytrium limacinum engineering bacteria for the KS gene knockout. According to the construction method of the schizochytrium limacinum engineering bacteria for the KS gene knockout, provided by the invention, schizochytrium limacinum recombinant bacteria for knocking out FabA (Beta-Hydroxydecanoyl-Acyl Carrier Protein-Dehydratase) genes are constructed; compared with a proportion of total oil and fat in original strains, the polyunsaturated fatty acid production ability of the engineering bacteria is reduced by 43 percent, and the production of C16 and C18 saturated fatty acids is increased by 29 percent.

Methods and means are provided to increase the oil content of plants, particularly oleaginous plants by preventing feedback inhibition by 18:1-Coenzyme A or 18:1-Acyl Carrier Protein of the acetyl CoA-carboxylase enzyme in cells of these plants in various manners.

The invention discloses a coupling object. The coupling object is obtained through the coupling of at least one protein in the four proteins of a recombined human epidermal growth factor and a recombined epidermal growth factor mutant with a carrier protein. The amino acid sequence of the recombined human epidermal growth factor is shown by the No.2 sequence in a sequence list. The recombined epidermal growth factor mutant can be one of the following proteins: 1)the protein obtained through the deletion of the No.53 arginine in the No.2 sequence in the sequence list; 2) the protein obtained through the deletion of the No.53 arginine and No.52 leucine in the No.2 sequence in the sequence list; and 3) the protein obtained through the addition of methionine before the No.1 asparagine in the No.2 sequence in the sequence list. The coupling object can be used as a vaccine for the active treatment of an epithelium sourced tumor after being mixed with an adjuvant. Results show that the vaccine can induct the high level antibodies in a mouse; the highest antibody titer reaches 1:20,000; and the prolonging rate of the survival period of the mouse reaches 69.4 percent.

The invention belongs to the technical field of drugs, relates to prodrugs designed on the basis of intestinal MCT1 (monocarboxylate transporter 1) carrier protein and a preparation method of the prodrugs, in particular to a preparation method of short-chain fatty acid analogs with MCT1 as a target. The preparation method comprises structural design and synthesis of the prodrug containing hydroxyl or amino anti-tumor drugs and modified with acetic acid, lactic acid and pyruvic acid analogs. According to a series of the prodrugs, oral bioavailability of the drug can be improved, and the release rate of the prodrugs can be increased. Involved derivatives represented by general formula (I) or (II) and pharmaceutically acceptable salts, hydrates, solvates or prodrugs of the derivatives have structures as follows in the description, wherein X, Y, n and Drug have definitions given in the description and claims.

The invention discloses a screening method of a bacterial enoyl acyl carrier protein (ACP) reductase inhibitor. The screening method comprises the steps: expressing fabK of enterococcus faecalis in escherichia coli, inhibiting and not inhibiting FabI of recombinant escherichia coli through triclosan, and constructing a double-plate sensitive difference screening model for FabK; constructing a fabI gene deletion mutant and a fabV gene deletion mutant of pseudomonas aeruginosa, and constructing a three-plate sensitive difference screening model for FabI and FabV on the basis of wild bacteria; screening a combination of the two screening models, and measuring half inhibition concentration of an activity extract on enoyl ACP reductase and detecting different expression strains of the enoyl ACP reductase to the activity extract, thus realizing secondary screening on sensitivity difference of the activity extract. The screening method disclosed by the invention has the beneficial effects that the screening method not only improves screening efficiency and simplifies screening of a broad-spectrum inhibitor, but also avoids construction of various drug screening models for diversity of the enoyl ACP reductase; and simultaneously, the method solves problems of high false positive and low screening efficiency.

The invention provides a synthetic method of an artificial florfenicol amine antigen and belongs to the technical field of biochemical engineering. Florfenicol reacts with 4-benzyl bromide, 20% H2SO4, K2CO3 and NH3.CH3OH to obtain a product provided with a carboxy group, the product is hapten, then the hapten is coupled with carrier protein by using a carbodiimide method, and the coupling ratio of a conjugate is determined by using an ultraviolet method. The artificial florfenicol amine antigen is successively synthetized, synthesis steps are simple and effective, the artificial florfenicol amine antigen can be completedly used for immunoassay, and a necessary artificial antigen is provided for people for future research.

The invention provides glycosylated polypeptide, a conjugate of the glycosylated polypeptide and carrier protein, as well as a preparation method and application of the glycosylated polypeptide. The glycosylated polypeptide has a sequential structure of Glyco-Val-His-Leu-Thr-Pro-Tyr-Ryr-Cys; the conjugate of the glycosylated polypeptide and the carrier protein is a product generated after the tail end Crs of the glycosylated polypeptide is coupled with the carrier protein (such as bovine serum albumin, myohemoglobin or hemocyanin); the glycosylated polypeptide or the conjugate of the glycosylated polypeptide and the carrier protein can be used for replacing an expensive pure human glycosylated hemoglobin antigen product to prepare a chromatographic testing bar (testing paper bar); the chromatographic testing b bar can be used for rapidly, accurately, quantitatively, simply and conveniently detecting the glycosylated hemoglobin in the whole blood.

The invention relates to the technical field of biology, in particular to a seed-specific promoter sequence separated from soybean and applications thereof. The nucleotide sequence is shown in SEQ No.1. The seed-specific promoter sequence is used as the seed-specific promoter of soybean. In the invention, the stearic acid-acyl carrier protein (ACP) desaturase promoter of soybean is cloned and the tissue expression characteristics and expression efficiency of the promoter in soybean are researched, thus an evidence for the effective use of the promoter is provided; and the promoter provides a good tool for the transgenic research of soybean and creates conditions for the researches and developments by utilizing the soybean seed as bioreactor.

The present disclosure pertains to methods of producing recombinant peptides that contain between 10 and 200 amino acid residues using novel carrier proteins derived from superfolder green fluorescent protein and its mutants.

The invention discloses a method for increasing fermentation yield of polyketone compounds. The method includes steps of: (1) culturing genetically engineered bacteria of an overexpressed acyl-coenzyme A synthesizing enzyme and an acyl carrier protein in a culture medium added with natural grease; and (2) extracting the polyketone compounds from a fermented product. The genetically engineered bacteria are a recombination expression transformant of the overexpressed acyl-coenzyme A synthesizing enzyme and the acyl carrier protein, which is prepared through conjugational transfer by an actinomycete onto recombinant escherichia coli containing a recombinant expression vector expressing the overexpressed acyl-coenzyme A synthesizing enzyme and the acyl carrier protein. In the method, an exogenetic acyl-coenzyme A synthesizing enzyme and an exogenetic acyl carrier protein are overexpressed in the bacteria generating the polyketone compounds through gene engineering technologies, so that by means of cheap natural grease as a raw material, the fermentation yield of the polyketone compounds is increased, wherein the method is never reported in references. The method is simple in operations, can provide precursors for synthesizing the polyketone compounds through the cheap natural grease, is low in cost and is convenient to popularize.

The invention discloses a 16-valent streptococcus pneumoniae conjugate vaccine composition. The vaccine composition is obtained in a manner that streptococcus pneumoniae capsular polysaccharide afterseparation and purification and carrier protein are subjected to coupling combination to be mixed, wherein the capsular polysaccharide includes 16 streptococcus pneumoniae serotypes of 1, 2, 3, 4, 5,6A, 6B, 7F, 9V, 11A, 14, 15B, 18C, 19A, 19F and 23F. The 16-valent streptococcus pneumoniae conjugate vaccine composition can cover most of common pathogenic bacterial types or drug-resistance bacterial types of our country, and has the broad-spectrum and efficient protection effect on high-risk susceptible populations such as new-born infants, old people and children under two years old.

The present disclosure pertains to methods of producing recombinant peptides that contain between 10 and 200 amino acid residues using novel carrier proteins derived from superfolder green fluorescent protein and its mutants.

The invention discloses a cyanobacteria aliphatic hydrocarbon key synthesis gene and application thereof. The aliphatic hydrocarbon synthesis gene includes the coding genes of fatty acyl-acyl carrier protein reductase (AAP) and fatty aldehyde deformylating oxygenase (ADO). The invention also relates a contractor, wherein the contractor comprises a promoter having activity and the aliphatic hydrocarbon key synthesis gene controlled by the promoter. By converting the contractor into escherichia coli, the genetic engineering escherichia coli can have an aliphatic hydrocarbon synthesis capacity. By converting the at least one contractor into the cyanobacteria, the aliphatic hydrocarbon yield of the genetic engineering cyanobacteria can be significantly improved. The cyanobacteria hydrocarbon production key gene disclosed by the invention can provide a new candidate gene resource for directly synthesizing aliphatic hydrocarbon novel bio-fuels from microorganisms.

The invention discloses a preparation method of a cell strain with sterol carrier protein (SCP) stable transformation of prodenia litura. The preparation method sequentially includes the steps of 1), cloning cDNA (complementary deoxyribonucleic acid) of SCP (single-cell protein)-2 gene from prodenia litura; 2), cloning the cDNA of the SCP-2 encoded protein into pcDNA5 / FRT plasmid to form recombined pcDNA5 / FRT-SCP2 plasmid; 3), applying the pcDNA5 / FRT-SCP2 plasmid to transform colon bacillus and obtaining transformed colon bacillus via cultivation; 4), extracting recombined plasmid DNA from the transformed colon bacillus and performing purification; 5), transfecting Chinese hamster ovary cells with purified and recombined pcDNA5 / FRT-SCP2 plasmid DNA; 6), selecting transfected CHO cells in a culture medium containing antibiotic hygromycinB and performing continuous subculture to obtain the cell strain. The invention aims to provide the cell strain with SCP stable transformation of the prodenia liture, the preparation method of the cell strain and an application of the cell strain. The cell strain has the advantages of high screening accuracy, simpleness in operation and accuracy in result.

The invention provides angiotensin II immunogen and a preparation method thereof. According to the angiotensin II immunogen and the preparation method thereof, when the N-end of angiotensin II is connected to carrier protein, a derivative of the angiotensin II is firstly synthesized, in the derivative of the angiotensin II, a segment of polypeptide is added to the N-end of the angiotensin II to serve as a connecting arm, the connecting arm comprises a flexible peptide, the flexible peptide is composed of Gly and Ser amino acid residues, the position of the Gly are distributed on the two sidesof the Ser, and therefore the torsion of the connecting arm is facilitated; and the derivative of the angiotensin II can effectively reduce the steric-hindrance effect of the carrier protein on smallmolecule hapten of the angiotensin II, and -OH on side chains of the Ser can improve the water solubility of molecules of the angiotensin II to a certain extent. The prepared angiotensin II immunogenhas the advantages of being simple in preparation method, low in cost, strong in immunogenicity and suitable for preparation of an angiotensin II antibody.