1351 results about "Metabolic pathway" patented technology

The present invention is generally directed to the evolution of new metabolic pathways and the enhancement of bioprocessing through a process herein termed recursive sequence recombination. Recursive sequence recombination entails performing iterative cycles of recombination and screening or selection to “evolve” individual genes, whole plasmids or viruses, multigene clusters, or even whole genomes. Such techniques do not require the extensive analysis and computation required by conventional methods for metabolic engineering.

Rice plants are disclosed with two separate, but synergistic mechanisms for resistance to herbicides that normally inhibit a plant's acetohydroxyacid synthase (AHAS) enzyme. The herbicide resistance of plants with both resistance mechanisms is substantially greater than one would expect from a simple combination of the two types of resistance. The first of the two resistance mechanisms is a metabolic pathway that is not fully understood, but that does not itself involve a mutant AHAS enzyme. The second resistance mechanism is a mutant AHAS enzyme, an enzyme that shows direct resistance to levels of herbicide that normally inhibit the enzyme, in both in vivo and in vitro assays. Besides controlling red rice, many AHAS-inhibiting herbicides also effectively control other weeds that are common in rice fields. Several of these herbicides have residual activity, so that a treatment controls both existing weeds as well as weeds that sprout later. No herbicide currently available for use on rice has residual activity against a broad spectrum of weeds including red rice. With effective residual activity against red rice and other weeds, rice producers now have a weed control system superior to those currently used.

The invention provides a group of relevant enzymes for preparing mannitol by performing anabolism on Chinese caterpillar fungus serving as a multifunctional production fungus and hirsutella sinensis based on glucose, a gene for encoding these enzymes and application thereof. The relevant enzymes include (1) hexokinase: manA1-A6 proteins of which the sequences are SEQ ID No.1-6, (2) phosphoglucoisomerase: manB1-B3 proteins of which the sequences are SEQ ID No.7-9, and (3) mannitol-1-P dehydrogenase: manC protein of which the sequence is SEQ ID No.10. In the invention, detailed researches are performed on the metabolic pathway of mannitol synthesized by using Chinese caterpillar fungus serving as a multifunctional production fungus, hirsutella sinensis and glucose on the aspect of principle, cloned DNA (Deoxyribose Nucleic Acid) comprising a nucleotide sequence provided by the invention can be transferred into engineering bacteria with transduction, conversion and conjugal transfer methods, and host mannitol is endowed with high expression by regulating the expression of a biosynthetic gene of the mannitol.

The present invention relates to 25 hitherto undescribed genes of B. licheniformis and gene products derived therefrom and all sufficiently homologous nucleic acids and proteins thereof. They occur in five different metabolic pathways for the formation of odorous substances. The metabolic pathways in question are for the synthesis of: 1) isovalerian acid (as part of the catabolism of leucine), 2) 2-methylbutyric acid and / or isobutyric acid (as part of the catabolism of valine and / or isoleucine), 3) butanol and / or butyric acid (as part of the metabolism of butyric acid), 4) propyl acid (as part of the metabolism of propionate) and / or 5) cadaverine and / or putrescine (as parts of the catabolism of lysine and / or arginine). The identification of these genes allows biotechnological production methods to be developed that are improved to the extent that, to assist these nucleic acids, the formation of the odorous substances synthesized via these metabolic pathways can be reduced by deactivating the corresponding genes in the micro-organism used for the biotechnological production. In addition, these gene products are thus available for preparing reactions or for methods according to their respective biochemical properties.

The present invention generally relates to a combination of the fields of tissue engineering, drug discovery and drug development. It more specifically provides new methods and materials for testing the efficacy and safety of experimental drugs, defining the metabolic pathways of experimental drugs and characterizing the properties (e.g., side effects, new uses) of existing drugs. Preferably, evaluation is carried out in three-dimensional tissue-engineered systems, wherein drug toxicity, metabolism, interaction and / or efficacy can be determined.

The present invention is generally provides recombinant microorganisms comprising engineered metabolic pathways capable of producing C3-C5 alcohols under aerobic and anaerobic conditions. The invention further provides ketol-acid reductoisomerase enzymes which have been mutated or modified to increase their NADH-dependent activity or to switch the cofactor preference from NADPH to NADH and are expressed in the modified microorganisms. In addition, the invention provides isobutyraldehyde dehydrogenase enzymes expressed in modified microorganisms. Also provided are methods of producing beneficial metabolites under aerobic and anaerobic conditions by contacting a suitable substrate with the modified microorganisms of the present invention.

The present invention provides for novel metabolic pathways to reduce or eliminate glycerol production and increase product formation. More specifically, the invention provides for a recombinant microorganism comprising a deletion of one or more native enzymes that function to produce glycerol and / or regulate glycerol synthesis and one or more native and / or heterologous enzymes that function in one or more engineered metabolic pathways to convert a carbohydrate source, such as lignocellulose, to a product, such as ethanol, wherein the one or more native and / or heterologous enzymes is activated, upregulated, or downregulated. The invention also provides for a recombinant microorganism comprising one or more heterologous enzymes that function to regulate glycerol synthesis and one or more native and / or heterologous enzymes that function in one or more engineered metabolic pathways to convert a carbohydrate source to ethanol, wherein said one or more native and / or heterologous enzymes is activated, upregulated or downregulated.

The methods described herein enable the evaluation of compounds on subjects to assess their therapeutic efficacy or toxic effects. The target of analysis is the underlying biochemical process or processes (i.e., metabolic process) thought to be involved in disease pathogenesis. Molecular flux rates within the one or more biochemical processes serve as biomarkers and are quantitated and compared with the molecular flux rates (i.e., biomarker) from control subjects (i.e., subjects not exposed to the compounds). Any change in the biomarker in the subject relative to the biomarker in the control subject provides the necessary information to evaluate therapeutic efficacy of an administered drug or a toxic effect and to develop the compound further if desired. In one aspect of the invention, stable isotope-labeled substrate molecules are administered to a subject and the label is incorporated into targeted molecules in a manner that reveals molecular flux rates through one or more metabolic pathways of interest. By this method, a comparison between subjects and control subjects reveals the effects of the chemical entity or entities on the biomarkers. This, in turn, allows for the identification of potential therapeutic uses or toxicities of the compound. Combinations of compounds can also be systematically evaluated for complementary, synergistic, or antagonistic actions on the metabolic pathways of interest, using the methods of the present invention as a strategy for identifying and confirming novel therapeutic or toxic combinations of compounds.

Compositions and methods useful for inducing an increase in fatty acid oxidation or mitochondrial biogenesis, reducing weight gain, inducing weight loss, or increasing Sirt1, Sirt3, or AMPK activity are provided herein. Such compositions can contain synergizing amounts of a sirtuin-pathway activators, including but not limited to resveratrol, in combination with beta-hydroxymethylbutyrate (HMB), keto isocaproic acid (KIC), leucine, or combinations of HMB, KIC and leucine.

The present invention relates to methods, systems and compositions, including genetically modified microorganisms, adapted to exhibit increased tolerance to 3-hydroxypropionic acid (3-HP), particularly through alterations to interrelated metabolic pathways identified herein as the 3-HP toleragenic pathway complex (“3HPTGC”). In various embodiments these organisms are genetically modified so that an increased 3-HP tolerance is achieved. Also, genetic modifications may be made to provide at least one genetic modification to any of one or more 3-HP biosynthesis pathways in microorganisms comprising one or more genetic modifications of the 3HPTGC.

Processes for biogenic production of a hydrogen-carbon-containing fluid from a hydrocarbon containing formation are described. The processes may include providing in the formation an anaerobic microorganism consortium containing one or more enzymes to activate a starting hydrocarbon by an addition of a chemical group to the hydrocarbon. The processes may further include converting the activated hydrocarbon into the hydrogen-carbon-containing fluid through one or more intermediate hydrocarbons, and recovering the hydrogen-carbon-containing fluid from the formation.

Aspects of the invention relate to methods for the production of difunctional alkanes in host cells. In particular, aspects of the invention describe components of genes associated with the difunctional alkane production from carbohydrate feedstocks in host cells. More specifically, aspects of the invention describe metabolic pathways for the production of adipic acid, aminocaproic acid, caprolactam, and hexamethylenediamine via 2-ketopimelic acid.

The present invention is directed to compounds, compositions and methods for treating or preventing viral infections using nucleoside analog monophosphate prodrugs. More specifically, HCV, Norovirus, Saporovirus, Dengue virus, Chikungunya virus and Yellow fever in human patients or other animal hosts. The compounds are certain 2,6-diamino 2-C-methyl purine nucleoside monophosphate prodrugs and modified prodrug analogs, and pharmaceutically acceptable, salts, prodrugs, and other derivatives thereof. In particular, the compounds show potent antiviral activity against HCV, Norovirus, Saporovirus, Dengue virus, Chikungunya virus and Yellow fever. This invention teaches how to modify the metabolic pathway of 2,6-diamino 2′-C-methyl purine and deliver nucleotide triphosphate(s) to polymerases at heretofore unobtainable therapeutically-relevant concentrations.

The present invention provides recombinant mircoorganisms comprising an isobutanol producing metabolic pathway and methods of using said recombinant microorganisms to produce isobutanol. In various aspects of the invention, the recombinant microorganisms may comprise a modification resulting in the reduction of pyruvate decarboxylase and / or glycerol-3-phosphate dehydrogenase activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

The invention is directed to materials and methods associated with polymorphic variants in two enzymes involved in folate-dependent and one-carbon metabolic pathways: MTHFD1 (5,10-methylenetetrahydrofolate dehydrogenase, 5,10-methenyltetrahydrofolate cyclohydrolase, 10-formyltetrahydrofolate synthetase) and methylenetetrahydrofolate dehydrogenase (NADP+dependent) 1-like (MTHFD1L). Diagnostic and therapeutic methods are provided involving the correlation of polymorphic variants in MTBFD1, MTHFD1, and other genes with relative susceptibility for various pregnancy-related and other complications.

The present invention provides recombinant microorganisms comprising isobutanol producing metabolic pathway with at least one isobutanol pathway enzyme localized in the cytosol, wherein said recombinant microorganism is selected to produce isobutanol from a carbon source. Methods of using said recombinant microorganisms to produce isobutanol are also provided. In various aspects of the invention, the recombinant microorganisms may comprise a cytosolically active isobutanol pathway enzymes. In some embodiments, the invention provides mutated, modified, and / or chimeric isobutanol pathway enzymes with cytosolic activity. In various embodiments described herein, the recombinant microorganisms may be microorganisms of the Saccharomyces clade, Crabtree-negative yeast microorganisms, Crabtree-positive yeast microorganisms, post-WGD (whole genome duplication) yeast microorganisms, pre-WGD (whole genome duplication) yeast microorganisms, and non-fermenting yeast microorganisms.

Lactobacillus plantarum having an anti-diabetic function and application thereof are disclosed, the strain is Lactobacillus plantarum NCU116, and the accession number is CGMCC 11885. The strain has a function to alleviate symptoms of diabetes, and the composition is in a form of food or a pharmaceutical composition. The present invention also discloses a method for screening and identification of the lactic acid bacteria strain as well as a method to achieve the anti-diabetic effect by regulating blood sugar, blood lipids and hormone levels and regulating metabolic pathways in the body. The daily-intake lactic acid bacteria or a composition thereof is used to replace a medicine to achieve the anti-diabetic effect; and the healthy daily-intake lactic acid bacteria or the composition thereof is free of toxic and side effects to human body is used as a new choice to alleviate the symptoms of diabetes.

Genetically engineered organisms for production of PHA copolymers containing 2-hydroxyacid monomers and the methods of making and using thereof have been developed. The copolymers containing 2-hydroxyacid monomers can be synthesized via biosynthesis by the action of a PHA polymerase in a living cell. By changing the genetic background of the cells, one can control specific metabolic pathways allowing control of the level of glycolic acid co-monomer in the PHA polymer.

The invention enables high-throughput screening of compounds in living systems to detect unanticipated or unintended biological actions. The invention also allows for screening, detection, and confirmation of new indications for approved drugs. Screening and detection of toxic effects of compounds also can be achieved by using the methods of the invention. The methods comprise administering isotope-labeled substrates to a living system so that the label is incorporated into molecules in a manner that reveals flux rates through metabolic pathways thought to be involved in a disease. Comparisons between living systems exposed to compounds and living systems not so exposed reveals the effects of the compounds on the flux rates through the metabolic pathways. Combinations or mixtures of compounds can be systematically screened to detect unanticipated or unintended biological actions, including synergistic actions, in the same manner.

Use of polymorphism site gene to predict ACEI medicine, its method and reagent kit are disclosed. It can be used to determine critical enzyme gene polymorphism site gene typing oligonucleotide and critical enzyme gene polymorphism site gene in cysteine metabolic pathway.

There are disclosed metabolically-engineered yeast and methods of producing n-butanol. In an embodiment, metabolically-engineered yeast is capable of metabolizing a carbon source to produce n-butanol, at least one pathway produces increased cytosolic acetyl-CoA relative to cytosolic acetyl-CoA produced by a wild-type yeast, and at least one heterologous gene encodes and expresses at least one enzyme for a metabolic pathway capable of utilizing NADH to convert acetyl-CoA to n-butanol. In another embodiment, a method of producing n-butanol includes (a) providing metabolically-engineered yeast capable of metabolizing a carbon source to produce n-butanol, at least one pathway produces increased cytosolic acetyl-CoA relative to cytosolic acetyl-CoA produced by a wild-type yeast, and at least one heterologous gene encodes and expresses at least one enzyme for a metabolic pathway utilizing NADH to convert acetyl-CoA to n-butanol; and (b) culturing the yeast to produce n-butanol. Other embodiments are also disclosed.

The present invention relates to combinatorial gene expression libraries and methods for making these. Such libraries are useful in discovery of novel and / or enhanced metabolic pathways leading to the production of novel compounds for e.g drug discovery and / or to the production of known compounds in novel quantities or in novel compartments of the cells. The expression libraries in particular are composed of host cells capable of co-ordinated and controllable expression of large numbers of heterologous genes in the host cells.

The embodiments described herein pertain to cells, and methods for preparing cells, that can be used as biocatalysts by altering enzymes that compete for a substrate or product of a pathway of interest such that the targeted enzyme is sensitive to a site-specific protease, which protease is expressed but relocated in the cell to a site where it is not in contact with the targeted enzyme in the intact cell. Upon cell lysis, the protease contacts the target enzyme, which is then inactivated by protease cleavage.

The metabolic processes involved in the formation of any glucose-based metabolite of a metabolic network are determined. A precursor molecule is labeled with a stable carbon (13C) isotope at specific positions. The label is allowed to distribute and rearrange in the system. Metabolites are recovered and analyzed against a control system to determine a set of metabolic pathway substrate fluxes caused by changes to the test system relative to the control system such as the addition of compound being tested as a potential drug.

The invention describes a process for preparing acetone starting from acetyl-coenzyme A comprising process steps A. enzymatic conversion of acetyl-CoA into acetoacetyl-CoA B. enzymatic conversion of acetoacetyl-CoA into acetoacetate and CoA and C. decarboxylation of acetoacetate to acetone and CO2, which is characterized in that the coenzyme A is not transferred in process step B to an acceptor molecule. In addition, process step B is surprisingly catalysed by enzymes of the classes of acyl-CoA thioesterase, acyl-CoA synthetase or acyl-CoA thiokinase.A completely novel metabolic pathway is concerned, because the enzymatic hydrolysis of acetoacetyl-CoA without simultaneous transfer of CoA to a receptor molecule has never previously been described for any microbial enzyme.

A process of increasing the cellular production of secondary metabolites, such as antibiotics, is provided. The process is particularly useful for increasing antibiotic production by bacterial cells, especially erythromycin. The process includes the step of increasing the activity of methylmalonyl-CoA mutase.

A method of doing business is disclosed whereby the process for selecting drug candidates is improved. The method involves the application of a technology which makes it possible to determine metabolic processes involved in the formation of any glucose-based metabolite. A precursor molecule is labeled with a stable carbon (13C) isotope at specific positions. The label is allowed to distribute and rearrange in the system. Metabolites are recovered and analyzed against a control system or known biochemical reactions and / or cycles to determine information such as metabolic pathway substrate flux caused by a compound acting on the system.

This invention describes genes, metabolic pathways, microbial strains and methods to produce methyl butanol and other compounds of interest from renewable feedstocks.

The methods of the present invention allow for the measurement of ribonucleotide reductase (RR) activity, an important enzyme in the de novo DNA synthesis pathway. Ribonucleotide reductase converts all four ribonucleotides to their deoxy form and is a rate-controlling step in this pathway. Biosynthetic pathways of deoxyribonucleotides (dN) have received considerable attention in the context of anti-proliferative chemotherapy. Inhibitors of various steps in dN biosynthesis, including inhibitors of RR are among the most useful chemotherapeutic agents in cancer, viral infections, and other therapeutic uses. DNA synthesis from the dN salvage pathway is also an important component to DNA replication. The relative contributions from RR vs. salvage pathways are critical to the actions and effectiveness of chemotherapeutic agents that act on nucleoside metabolic pathways. Until now, however, it has not been possible to study these metabolic processes in vivo. Disclosed within are methods of measuring RR activity in vivo and in vitro which find use, among other things, in drug discovery, development, and approval.

The invention discloses a genetically engineered bacteria used for producing stevia glycosyltransferase UGT76G1; a UGT76G1 coding gene is inserted between the restriction enzyme cutting sites EcoRI and XhoI of a PYes2 carrier, so as to construct a recombinant plasmid, and then the recombinant plasmid is introduced to expression host Saccharomyces cerevisiae YPH499 to obtain the engineered bacteria; and the coding gene of the UGT76G1 is GenBank, No. GenBank: AY345974.1, and the gene sequence of the coding gene is named as UGT (Udp Glucuronyl Transferase). The invention also discloses a construction method of the genetically engineered bacteria and the application of the bacteria to the production of rebaudioside A. According to the invention, under the condition that expensive UDPG (Uridine Diphosphate Glucose) is not added, cheap carbon source glucose is used as a substrate, the metabolic pathway of UDPG in the yeast is regulated, and then the rebaudioside A is produced from St glycosides through whole cell catalysis.