145 results about "De novo synthesis" patented technology

The invention discloses a biological production method of tanshinol, which comprises the following steps: synthesizing p-hydroxyphenylpyruvic acid into 3,4-dihydroxyphenylpyruvic acid under the catalytic action of a p-hydroxyphenylacetic acid meta-position hydroxylation enzyme, and then, synthesizing to obtain the tanshinol under the catalytic action of a D-lactate dehydrogenase; or synthesizing p-hydroxyphenylpyruvic acid into p-hydroxyphenyllactic acid under the catalytic action of a D-lactate dehydrogenase, and then, synthesizing to obtain the tanshinol under the catalytic action of a p-hydroxyphenylacetic acid meta-position hydroxylation enzyme. According to the invention, gene engineering glutamic acid corynebacteria and Escherichia coli are used to produce the tanshinol through fermentation, and the tanshinol can be synthesized without adding a substrate, thereby realizing de novo synthesis of the tanshinol and solving the problem on the source of the tanshinol; and meanwhile, the production cost is lowered to the greatest extent. Thus, the biological production method is beneficial to industrial production.

The present invention relates to the recombinant method used for the production of soluble form of an antibody that binds to CD20 for treatment of patients with relapsed or refractory, low-grade or follicular, CD20-positive, B-cell non-Hodgkin's lymphoma (NHL). The treatment will comprise the use of immunologically active anti-CD20 antibodies; or radiolabeled anti-CD20 antibodies and or cooperative strategies where both labeled and non-labeled antibodies will be used for treatment of NHL. The procedure describes the de novo synthesis of the nucleic acid sequence encoding anti-CD20, transformation of the constructed nucleic acid sequences into competent bacteria and the sub-cloning of the same into mammalian expression vectors for expression of the desired protein. DNA constructs comprising the control elements associated with the gene of interest has been disclosed. The nucleic acid sequence of interest has been codon optimized to permit expression in the suitable mammalian host cells.

RTP801 represents a unique gene target for hypoxia-inducible factor-1 (HIF-1). Down-regulation of the mTOR pathway activity by hypoxia requires de novo mRNA synthesis and correlates with increased expression of RTP801. The present invention relates to screening systems utilizing RTP801 and/or RTP801 interactors and/or RTP801 biological activity, to drug candidates identified by such screening systems, and to the use of such drug candidates in the treatment of various disorders.

Compositions containing epidithiodioxopiprazines and methods of their use are provided. Epidithiodioxopiprazines can be isolated from natural resources or synthesized de novo. Moreover, epidithiodioxopiprazines, including Verticillin A, are shown to effectively sensitize multiple types of tumor cells to TRAIL-induced apoptosis. In addition, epidithiodioxopiprazines, including Verticillin A, are shown to effectively overcome cancer cell resistance to existing drugs (i.e. Etoposide, Cisplatin, 5-FU and Doxorubicin). Therefore, compositions and methods are provided for use in sensitizing target cancer cells to death receptor- and other anticancer drugs-induced apoptosis. Methods of treating cancer in a subject in need thereof are also provided.

The invention discloses a biological method for producing resveratrol, which comprises the following steps: ligating target genes obtained by restriction enzyme digestion to an expression vector, wherein the target genes include the sequences of phenylalanine hydroxylase (PAH), tyrosine ammonia lyase (TAL) 4-coumarate:coenzyme A ligase (4CL) and resveratrol synthase (RS); transforming the constructed expression vector into a strain to obtain a recombinant engineered strain; and fermenting the recombined engineered strain. Compared with the prior art, the technical scheme provided by the invention uses the genetically engineered strain for fermentation to produce resveratrol, so as to realize denovo synthesis of resveratrol with no need of adding substrates. The method disclosed by the invention solves the source problem of resveratrol, and at the same time, reduces the production cost in a maximum extent, and is beneficial for industrial production.

The invention relates to a recombinant expression plasmid vector for producing fucose-based lactose, metabolic engineering bacteria, and a production method and belongs to the fields of metabolic engineering, food fermentation and the like. In the invention, genes for encoding GDP-mannose-6-dehydrogenase, GDP-fucose synthetase, lactose permease, and alpha-1,2-fucose transferase or alpha-1,3-fucosetransferase, which are required in a de-novo synthesis route of the fucose-based lactose, are expressed in corynebacterium glutamicum, so that recombinant corynebacterium glutamicum is constructed toachieve synthesis of 2'-fucose-based lactose or 3'-fucose-based lactose. In addition, by over-expression of the genes for encoding phosphomannose isomerase, phosphomannose mutase, and mannose-1-phosphate guanylyltransferase in the corynebacterium glutamicum, high yield of the fucose-based lactose is achieved. According to the method, the engineering bacteria can synthesize the fucose-based lactose by means of glucose or glycerol, has advantages of high growth speed and good safety, and has significant industrial potential.

The present invention discloses a construction method of escherichia coli for synthesizing fucosylated oligosaccharide by fermentation. The construction method includes the steps: (1) overexpressing at least one gene for encoding enzymes required for denovo synthesizing GDP-L-fucose in a prokaryotic host cell; (2) expressing an exogenous gene for encoding a fucosyltransferase in the prokaryotic host cell; (3) reducing or eliminating the activity of GDP-mannanohydrolase in the prokaryotic host cell; and (4) reducing or eliminating the activity of beta-galactosidase in the prokaryotic host cell.According to the present invention, the escherichia coli is constructed by using the method, and the escherichia coli may be used for preparing the fucosylated oligosaccharide. Shown by a result generated by performing fermentation production verification on the fucosylated oligosaccharide (with 2'-fucosyllactose as an example) in a 5L tank by adopting an engineering strain constructed by the present invention, the highest production level of the 2'-fucosyllactose (2'-FL) may reach 50 g/L.

The invention relates to recombinant escherichia coli for producing 2'-fucosyllactose and application thereof, and belongs to the field of microbial metabolic engineering. According to a regulation and control strategy for the expression level of 2'-fucosyllactose de novo synthesis route key enzyme, different enzymes are subjected to regulation and control of genome overexpression or plasmid overexpression, the efficiency of a metabolic pathway can be greatly improved, compared with the prior art, the efficiency of a genetic engineering strain constructed through related strategies is improvedby 13 times or above, and a foundation is laid for industrial production of the 2'-fucosyllactose.

Described is a method for decreasing the amount of mSREBP in a cell characterized by an elevated level of mSREBP comprising contacting the cell with an agent that specifically inhibits de novo synthesis of ceramide in the cell, thereby decreasing the amount of mSREBP in the cell. Also described are related methods and articles of manufacture.

Metabolic auxotroph of Vibrio cholerae 0139 synonym Bengal which has a mutation in its hem A gene and which is not capable of synthesizing aminolevulinic acid (ALA) de novo and which is obtained from a parent strain originally isolated from a patient's coproculture having all the identifying characteristics of Vibrio cholerae 0139 synonym Bengal is described. In this strain the hem A gene is mutated by inserting a kanamycin resistant gene cassette. Another metabolic auxotroph of Vibrio cholerae 01 El Tor where the hem A gene is mutated by a frame shift mutation is disclosed. Methods of producing the strains are disclosed.

The invention belongs to the technical field of biological engineering, and relates to the construction, strain, and application of a microbial fuel cell electricity-producing genetically engineered bacteria. With a molecular biology method, a common key gene nadE in Escherichia coli NAD<+> de-novo synthesis pathway and anaplerotic pathway is heterologously expressed in pseudomonas aeruginosa PAO1, such that the high-electricity-production genetically engineered strain pseudomonas aeruginosa PA-nadE is obtained, and can be applied in the development of the field of microbial fuel cell. Also, good basis is provided for the genetic engineering researches of electricity-producing pseudomonas aeruginosa.

The invention provides a recombinant escherichia coli for synthesizing 2'-fucosyllactose and construction method thereof, and the recombinant Escherichia coli is obtained by knocking out a beta-galactosidase gene in the genome of Escherichia coli, synthesizing a key enzyme gene of colanic acid from guanosine diphosphate fucose, enhancing expression key enzyme genes of lactose permease genes and a guanylate diphosphate fucose denovo synthesis route and expressing exogenous fucosyltransferase genes. According to the scheme, lactose hydrolysis can be reduced, and the rate of transferring exogenous lactose into cells is increased, so that the concentration of lactose in the cells is effectively increased; meanwhile, the consumption of the guanosine diphosphate fucose is reduced, and the concentration of the guanosine diphosphate fucose in cells is effectively improved; through codon optimization, the expression of an exogenous fucosyltransferase gene is improved, and synthesizing of the 2'-fucosyllactose is effectively promoted.

The invention provides novel L-proline-4-hydroxylase, amino acid sequence of which is as shown in the SEQ ID No.2. The L-proline-4-hydroxylase is derived from Micromonospora sp. CNB394. The inventionalso provides genetic engineering bacterium E. coli HYP for high-efficiency expression of the above L-proline-4-hydroxylase and denovo synthesis of high-yield trans-4-hydroxy-L-proline. The genetic engineering bacterium has advantages of good genetic stability and strong fermentation and acidogenesis capability, and contains no plasmid. By genome expression of the above novel L-proline-4-hydroxylase and using a cheap carbon source such as glucose and the like as a substrate, efficient denovo synthesis of trans-4-hydroxy-L-proline is achieved. After fermentation for 42 h, yield of the trans-4-hydroxy-L-proline reaches up to 51 g/L. The invention has a good industrial application prospect.

The present invention relates to a method of treating or preventing cellular dysfunction and death caused by genetic, degenerative, toxic, traumatic, ischemic, infectious, neoplastic and inflammatory diseases and aging—and their neurological symptoms and manifestations, which includes administering d-methadone, beta-d-methadol, alpha-l-methadol, beta-l-methadol, alpha-d-methadol, acetylmethadol, d-alpha-acetylmethadol, l-alpha-acetylmethadol, beta-d-acetylmethadol, beta-l-acetylmethadol, d-alpha-normethadol, l-alpha normethadol, noracetylmethadol, dinoracetylmethadol, methadol, normethadol, dinormethadol, EDDP, EMDP, d-isomethadone, normethadone, N-methyl-methadone, N-methyl-d-methadone, N-methyl-l-methadone, l-moramide, levopropoxyphene, pharmaceutically acceptable salts, or mixtures thereof, including deuterated and tritium analogues, whether isolated from its enantiomer or synthesized de novo.

The present invention relates to the recombinant method used for the production of soluble form monoclonal antibody that binds to CD52. The procedure describes the de novo synthesis of the nucleic acid sequence encoding anti-CD 52, transformation of the constructed nucleic acid sequences into competent bacteria and the sub-cloning of the same into mammalian expression vectors for expression of the desired protein. DNA constructs comprising the control elements associated with the gene of interest has been disclosed. The nucleic acid sequence of interest has been codon optimized to permit expression in the suitable mammalian host cells.

The invention belongs to the technical field of immunological test, and in particular relates to a method for preparing Sudan 1 capable of being directly marked and application of the Sudan 1. In the method, a technical route of denovo synthesis is mainly utilized, amino active group is introduced successfully on the premise of not changing an antigenic determinant in a host molecular structure of the Sudan 1, and a trim capable of directly being connected with fluorescent substances and enzymes is provided for the Sudan 1; and an important problem of competitive antigen preparation is solved for establishing a Sudan 1 competitive method enzyme-linked immuno sorbent assay (ELISA) detection system. The whole method is simple and practicable, easy to operate, and high in efficiency of connecting the fluorescent substances and the enzymes.