554 results about "Synthetic enzyme" patented technology

The present invention features methods and compositions for preventing, reducing, or treating a traumatic, metabolic or toxic peripheral nerve lesion or pain including, for example, neuropathic pain, inflammatory and nociceptive pain by administering to a mammal in need thereof a compound that reduces the expression or activity of BH4. According to this invention, this reduction may be achieved by reducing the enzyme activity of any of the BH4 synthetic enzymes, such as GTP cyclohydrolase (GTPCH), sepiapterin reductase (SPR), or dihydropteridine reductase (DHPR); by antagonizing the cofactor function of BH4 on BH4-dependent enzymes; or by blocking BH4 binding to membrane bound receptors. The compounds of the invention may be administered alone or in combination with a second therapeutic agent. The invention also provides methods for diagnosing pain or a peripheral nerve lesion in a mammal by measuring the levels of BH4 or its metabolites in biological sample. Alternatively, pain or a peripheral nerve lesion may be diagnosed by measuring the levels or activity of any one of the BH4 synthetic enzymes in tissue samples of a mammal. Also disclosed are screening methods that make use of BH4 or BH4 synthetic enzymes, BH4-dependent enzymes, and BH4-binding receptors for the identification of novel therapeutics for the treatment, prevention, or reduction of pain.

Compositions and methods for producing fatty acid derivatives using recombinant microorganisms are described herein.

The present invention concerns a novel means by which specific chosen reactions can be accelerated through the use of a new type of artificial enzyme. The invention allows specific reactions to occur at an accelerated rate, even in the presence of other non-chosen molecules, which may be very similar in structure to the chosen reactant. The reactions may be stoichiometric or catalytic.

The present invention relates to a kit for the treatment of cancer comprising (a) a container for containing a first compound (i) or a precursor thereof, said first compound or precursor being a compound that oxidizes glutathione (GSH); (b) a container for containing a second compound (ii) or a precursor thereof, said second compound or precursor being a compound that forms an adduct or conjugate with GSH; (c) a container for containing a third compound (iii) or a precursor thereof, said third compound or precursor being a compound that inhibits the rate-limiting enzyme of GSH biosynthesis, gamma-glutamylcysteine synthetase (GCS); and (d) a container for containing a fourth compound (iv) or a precursor thereof, said fourth compound or precursor being a compound that inhibits the enzyme responsible for the conversion of GSSG to GSH, glutathione reductase (GR).

The present invention is in the field of bioelectricity. The present invention provides energy generating systems, methods, and devices that are capable of converting chemical energy stored in sugars into useful electricity.

A unique carotenogenic biosynthetic gene cluster has been isolated from Panteoa agglomerans strain DC404, wherein the genetic organization of the cluster is crtE-idi-crtY-crtI-crtB-crtZ. The genes contained within this cluster encode geranylgeranyl pyrophosphate (GGPP) synthetase (CrtE), isopentenyl pyrophosphate isomerase (Idi), lycopene cyclase (CrtY), phytoene desaturase (CrtI), phytoene synthase (CrtB), and β-carotene hydroxylase (CrtZ). The gene cluster, genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

The invention relates to orthogonal pairs of tRNAs and aminoacyl-tRNA synthetases that can incorporate the coumarin unnatural amino acid L-(7-hydroxycoumarin-4-yl) ethylglycine into proteins produced in eubacterial host cells such as E. coli. The invention provides, for example but not limited to, novel orthogonal synthetases, methods for identifying and making the novel synthetases, methods for producing proteins containing the unnatural amino acid L-(7-hydroxycoumarin-4-yl)ethylglycine and related translation systems.

The invention discloses a recombinant pichia pastoris engineering bacterium and a method for using the recombinant pichia pastoris engineering bacterium as a whole-cell catalyst for synthesizing RA (rebaudioside A). The recombinant pichia pastoris engineering bacterium is obtained through conversion of pichia pastoris after linearization of an expression vector containing exogenous DNA as follows: the DNA sequence for coding sucrose synthetase Sus1 is represented by SEQ ID NO.3, and the DNA sequence for coding UDP-glycosyl transferase UGT76G1 is represented by SEQ ID NO.4. The intracellular expression of the Sus1 and the surface expression of the UGT76G1 are realized by the aid of the recombinant pichia pastoris engineering bacterium, and the recombinant pichia pastoris engineering bacterium is used as the whole-cell catalyst and can be effectively used for synthesizing the RA, so that the synthetic efficiency of the RA is improved, the raw material utilization rate is increased, the production cost is reduced, and a new industrial way is opened for production and processing of the RA.

A method for producing lacto-N-biose I and galacto-N-biose inexpensively and conveniently is provided. The method for producing lacto-N-biose I or galacto-N-biose, characterized in that the method comprises causing: (i) a combination of a carbohydrate raw material with an enzyme that catalyzes phosphorolysis of the carbohydrate raw material to give a-glucose-1-phosphate; and (ii) a combination of an enzyme that converts α-glucose-1-phosphate to UDP-glucose and an enzyme that converts UDP-galactose to galactose-1-phosphate with their cofactors, and/or a combination of an enzyme (UDP-Gly synthase) that converts α-glucose-1-phosphate and UDP-galactose to UDP-glucose and α-galactose-1-phosphate, respectively, with its cofactor(s) to act in the presence of N-acetylglucosamine or N-acetylgalactosamine, phosphoric acid, lacto-N-biose phosphorylase (EC 2.4.1.211), and UDP-glucose-4-epimerase (EC 5.1.3.2).

This disclosure concerns compositions and methods for genetically encoding and expressing prokaryotic tRNAs in eukaryotic cells. In some embodiments, the disclosure concerns methods and compositions for expressing unnatural amino acids in eukaryotic cells using orthogonal tRNA/synthetase pairs. In certain embodiments, the methods involve expressing prokaryotic tRNA/synthetase pairs in eukaryotic cells, for instance mammalian cells or yeast cells (such as those that are NMD-deficient), under the control of a pol III promoter, for instance a type-3 pol III promoter or an internal leader promoter. Also provided are cell lines that are NMD-deficient and methods of increasing the efficiency of UAA incorporation in a cell that include de-activating the NMD pathway in the cell. Also provided are methods increasing the efficiency of incorporation of an unnatural amino acid in a cell by disrupting a Nonsense-Mediated mRNA Decay—(NMD) pathway in the cell.

The invention discloses a genetically engineered bacterium for producing L-arginine and a construction method and application thereof. According to the invention, a gene for encoding carbamyl phosphate synthetase and a gene for encoding L-arginine biosynthetic pathway enzyme are integrated in escherichia coli; a synthetic route of arginine in escherichia coli and metabolic flux related to argininein a whole amino acid metabolic network are analyzed and reconstructed to obtain genetically engineered bacteria which are clear in genetic background, do not carry plasmids, are not mutated and canstably and efficiently produce L-arginine; and the genetically engineered bacteria have good L-arginine production capacity.

A human-derived novel chondroitin synthase, which is an enzyme for synthesizing a fundamental backbone of chondroitin and has both glucuronic acid transferring activity and N-acetylgalactosamine transferring activity.

The invention relates to a process for preparing silk functional fabric, which grafts the cyclodextrin onto the silk fiber through the chemical bond to obtain the silk functional fabric product with cyclodextrin solid chirality hydrophobic cavity, wherein the process can be used for preparing fabric with aromatic and medical health care functions and also can be used in the fields of molecule identification, waste water purification, artificial enzyme system forming and pharmaceutical slow releasing.

The invention discloses a gene for coding a glutamine dipeptide biosynthetic enzyme and application thereof. The nucleotide sequence of the gene is shown as SEQ ID NO.1. The invention further discloses an amino acid sequence coded by the gene, and provides a recombinant vector containing the gene, recombinant escherichia coli, and a method for performing biotransformation to synthesize the glutamine dipeptide by using the gene. The gene and the application of a recombinant bacterial strain of the gene have the advantages of high mol conversion rate, high reaction speed, easy separation, low cost and the like. In the synthesis method, the maximum mol conversion rate of the glutamine dipeptide can reach 83.3 percent; meanwhile, the thalli can be applied to the catalytic synthesis of the glutamine dipeptide again after the circulation recovery; in addition, the catalytic activity is stable. Therefore high market competitive power and application values are realized; a foundation is laid for the industrial production of the glutamine dipeptide.

The present invention provides cloning sequencing, analyzing, function research of a biosynthesis gene cluster of an antibiotic-Azinomycin B having antitumor activity produced by streptomyces, and its application. The whole gene cluster includes 34 genes: one repeatedly using I type polyketide synthase gene; two naphthalene ring modification enzyme genes; 8 non-ribosomal polypeptide skeleton synthesis and modification enzyme genes; 11 non-natural amino acid structure unit synthase genes; 1 resistance gene; 3 post modification enzyme genes and 8 genes which functions are not determined. The genetic operation of the biosynthesis gene breaks the synthesis of Azinomycin B; the precursor compound is produced by the heterologous expression of synthesis gene and modification gene of naphthalene ring. The gene of the invention and the protein can be used for searching and finding compound or gene, protein applied in medical, industry or agriculture.

The invention discloses an enzyme biological fuel cell positive pole and a preparation method and application thereof. Carbon paper is served as an electrode substrate, gelatin is served as modified material of a carbon nano tube, the carbon nano tube with the gelatin modified and bovine serum albumin are served as enzyme carriers, enzyme is served as catalyst, glutaraldehyde is served as cross-linking agent, and therefore the enzyme biological fuel cell positive pole is combined. The preparation method is simple and effective, convenient to operate, environment-friendly and low in cost. Due to the fact that the gelatin can successfully solve the problem that the carbon nano tube is not good in dispersibility in water solution, the enzyme biological fuel cell positive pole which is provided by the preparation method can greatly improve power density of the enzyme biological fuel cell positive pole. The enzyme biological fuel cell positive pole can be used widely, not only in preparation of biological electrodes of enzyme biological fuel cells, but also in preparation of electrodes of biological sensors.

The present invention provides novel orthogonal suppressor tRNAs, aminoacyl-tRNA synthetases, and suppressor tRNA/aminoacyl-tRNA synthetase pairs. In preferred embodiments the suppressor tRNAs function in mammalian cells and in certain embodiments they are not aminoacylated by any of the mammalian cytoplasmic aminoacyl-tRNA synthetases. The invention provides a novel ochre suppressor tRNA that fulfills these criteria. The invention further provides novel amber and opal suppressor tRNAs having a range of different translation efficiencies. The invention also provides mammalian cells containing the suppressor tRNAs, aminoacyl-tRNA synthetases, or both. The suppressor tRNAs may be introduced into the cell from the exterior or may be expressed by the cell. They may be aminoacylated with either a natural or an unnatural amino acid. The suppressor tRNAs, aminoacyl-tRNA synthetases, or both, may be expressed by a mammalian cell in a regulated manner. The invention further provides methods for synthesizing proteins using the inventive suppressor tRNAs and aminoacyl-tRNA synthetases and proteins produced by the methods.

The invention discloses recombinant corynebacterium glutamicum for producing gamma-polyglutamic acid as well as a construction method and a use of the recombinant corynebacterium glutamicum. The recombinant corynebacterium glutamicum is characterized by being an engineering bacteria which is obtained by transferring a recombinant expression plasmid containing a gamma-PGA (Poly Glutamic Acid) synthetase composite gene pgsBCA to an original strain wild C.glutamicum ATCC13869. The recombinant corynebacterium glutamicum disclosed by the invention contains the gamma-PGA synthetase composite gene pgsBCA, so that much gamma-PGA can be synthesized under biotin limit or under the fermentation condition of adding Tween40, therefore, the glutamic acid adding cost in the gamma-PGA production process is reduced, and a foundation is laid for the innovation in realizing the one-step production of gamma-PGA better.

Isolated mogroside and mogrol biosynthetic pathway enzyme polypeptides useful in mogroside biosynthesis are provided. Mogroside biosynthetic pathway enzymes of the invention include squalene epoxidase (SE), expoxy hydratase (EH), cytochrome p450 (Cyp), cucurbitadienol synthase (CDS) and udp-glucosyl-transferase (UGT). Also provided are methods of producing a mogroside using the isolated mogroside and mogrol biosynthetic enzyme polypeptides, the methods comprising contacting a mogrol and/or a glycosylated mogrol (mogroside) with at least one UDP glucose glucosyl transferase (UGT) enzyme polypeptide of the invention catalyzing glucosylation of the mogrol and/or the glucosylated mogrol to produce a mogroside with an additional glucosyl moietie(s), thereby producing the mogroside. Alternatively or additionally provided is a method of synthesizing a mogrol, the method comprising contacting a mogrol precursor substrate with one or more mogrol biosynthetic pathway enzyme polypeptides as described herein catalyzing mogrol synthesis from the mogrol precursor substrate, thereby synthesizing the mogrol.

The invention provides a glutathione synthetase-system recombinant plasmid as well as a construction method and application thereof, a recombinant strain for expressing glutathione synthetase systems as well as a construction method and application thereof, and a method for producing glutathione. The glutathione synthetase-system recombinant plasmid comprises a GSH1 sequence, a GSH1 sequence and a suitable vector fragment. The recombinant strain for glutathione synthetase systems is constructed by use of the glutathione synthetase-system recombinant plasmid. The method for producing glutathione comprises the step of fermenting the recombinant strain for glutathione synthetase systems. Glutathione-producing Pichia pastoris can be constructed by use of the coexpression plasmid provided by the invention; the obtained glutathione-producing Pichia pastoris is fermented and cultured; and the yield of glutathione in obtained fermentation broth is significantly higher than the current reported highest level of glutathione fermentation production. The invention has the advantages of improving the utilization rate of raw materials and reducing production cost and energy consumption, and can be used for industrial production.

The invention relates to a biosynthesis method of dammarenediol and provides a producing strain of the dammarenediol. A pichia pastoris engineering strain, which can recombine and express a dammarenediol synthetic enzyme with high efficiency, is constructed through a genetic engineering technical means, a precursor for synthesizing protopanaxadiol, namely the dammarenediol is further synthesized, and a foundation is finally laid for biosynthesis and large-scale industrial production of the protopanaxadiol.

A genetic recombinant method of pseudomonas aeruginosa for high-yield producing rhamnolipid includes following steps: (1) designing a primer for amplifying RhlAB gene according to the RhlAB gene sequence of rhamnolipid synthetase; (2) with chromosome of wild pseudomonas aeruginosa producing rhamnolipid as a template, performing PCR amplification to the RhlAB gene and performing rubber cutting purification to target genes through a purification kit; (3) performing enzyme digestion to the purified RhlAB gene to introduce the purified RhlAB gene into an escherichia coli-pseudomonas aeruginosa shuttle expression vector pAK1900 to obtain new plasmid pAK1900-AB; (4) introducing the constructed shuttle expression vector pAK1900-AB into a recipient bacterium pseudomonas aeruginosa competent cell through a heat-shock conversion method, coating a carboxyl-benzyl resistance plate with the competent cell and performing PCR verification to the constructed engineering bacteria; and (5) performing fermentation in a shake flask and measuring the yield of the rhamnolipid in a sulfuric acid-anthranone manner. The method can greatly reduce the production cost of the rhamnolipid, provides basis for industrial application of the rhamnolipid and can promote high-yield production of the rhamnolipid.