274 results about "Monooxygenase" patented technology

The present invention relates to co-administering a Hepatitis C virus NS3/4A protease inhibitor and a cytochrome P450 monooxygenase inhibitor. The combination acts by interfering with the life cycle of the hepatitis C virus and is therefore useful as an antiviral therapy. As such, the combination may be used for treating or preventing Hepatitis C infections in patients. The invention also relates to compositions comprising the combination of inhibitors. The invention also relates to kits and pharmaceutical packs comprising a Hepatitis C virus NS3/4A protease inhibitor and a cytochrome P450 monooxygenase inhibitor. The invention also relates to processes for preparing these compositions, combinations, kits, and packs.

The invention provides a modified variant of dicamba monooxygenase (DMO). The invention relates to the unexpected finding that cells expressing this DMO exhibit high levels of tolerance to the herbicide dicamba. Compositions comprising DMO-encoding nucleic acids and methods of use are provided.

The invention provides for identification and use of certain chloroplast transit peptides for efficient processing and localization of dicamba monooxygenase (DMO) enzyme in transgenic plants. Methods for producing dicamba tolerant plants, methods for controlling weed growth, and methods for producing food, feed, and other products are also provided, as well as seed that confers tolerance to dicamba when it is applied pre- or post-emergence.

A full length choline monooxygenase (CMO) cDNA was cloned from spinach and used to transform plants which do not naturally express CMO. A method is presented to improve stress tolerance of crops following engineering of CMO and BADH in plants that lack glycine betaine accumulation. Also provided are fragments useful as probes to isolate other CMO-type genes, and antisense sequences which inhibit the production of CMO. Reduction of glycine betaine as a consequence of antisense expression of CMO in species naturally accumulating glycine betaine, improves the transgenic plant's tolerance toward pathogens and pests and/or enhances its nutritional quality.

The present invention relates generally to novel genetic sequences which encode fatty acid epoxygenase enzymes. In particular, the present invention relates to genetic sequences which encode fatty acid DELTA12-epoxygenase enzymes comprising mixed function monooxygenase enzymes. More preferably, the present invention provides cDNA sequences which encode plant fatty acid epoxygenases, in particular the Crepis palaestina DELTA12-epoxygenase and homologues, analogues and derivatives thereof. The genetic sequences of the present invention provide the means by which fatty acid metabolism may be altered or manipulated in organisms such as yeasts, moulds, bacteria, insects, birds, mammals and plants, in particular to convert unsaturated fatty acids to epoxygenated fatty acids therein. The invention extends to genetically modified oil-accumulating organisms transformed with the subject genetic sequences and to the oils derived therefrom. The oils thus produced provide the means for the cost-effective raw materials for use in the efficient production of coatings, resins, glues, plastics, surfactants and lubricants, amongst others.

The present invention provides methods and DNA constructs for the genetic engineering of plant cells to produce plants which produce substantially seedless fruit in the absence of exogenous growth factors (auxins or cytokinins) and in the absence of pollination. The substantially seedless fruits produced by the methods described herein are about the size of wildtype seeded fruit (or somewhat larger) and these fruits are equal to or superior to the wildtype seeded fruit with respect to solid content and flavor. The seedless fruits of the present invention are produced in transgenic plants which contain and express auxin or cytokinin biosynthetic genes, e.g., tryptophan oxygenase or isopentenyl transferase coding sequences expressed under the regulatory control of GH3 or AGL promoter sequences directing preferential or tissue specific expression of a downstream gene in the ovaries or developing fruit.

The present invention relates to novel monooxygenase nucleic acids and polypeptides created using mutagenesis, DNA shuffling, or both, in a single iteration or multiple iterations, and methods for their creation and use. The monooxygenase enzymes of the present disclosure have particular utility as biocatalysts in industrial chemical redox reactions, such as the oxidation of aromatic hydrocarbons, for example, toluene, benzene, or nitrobenzene, into industrially desirable products. The systems and processes of the present invention are especially useful for the coupled synthesis and recovery of catechols, methylcatechols, resorcinols, methylresorcinols, hydroquinones, methylhydroquinones, hydroxybenzenes, cresols, nitrobenzenes, and nitrohydroxyquinones.

The invention discloses recombinant saccharomyces cerevisiae for producing dammarenediol and protopanoxadiol using xylose and a construction method. The construction method comprises the steps of replacing a promoter of a saccharomyces cerevisiae xylulokinase gene XKS1 with a promoter PFBA1 by virtue of a homologous recombination method, introducing xylose reductase XYL1 and a xylitol dehydrogenase XYL2 expression cassette, increasing the activities of transketolase TKL1 and transaldolase TAL1 so as to obtain recombinant bacteria 1, introducing a farnesyl-diphosphate farnesyltransferase gene ERG9, a squalene monooxygenase gene ERG1 and a dammarenediol synthase gene DS into the recombinant bacteria 1 so as to obtain recombinant bacteria 2, and introducing a nicotinamide adenine dinucleotide-hydroxymethylglutaryl coenzyme A reductase gene NADH-HMGr, farnesyl diphosphatesynthase ERG20 and a protopanoxadiol synthase-cytochrome P450 reductase fusion protein gene PPDS-ATR1 into the recombinant bacteria 2, so as to obtain recombinant bacteria 3. According to the recombinant saccharomyces cerevisiae, dammarenediol and protopanoxadiol can be artificially synthesized by virtue of xylose.

Conversion in vitro of X-Gly to X-alpha-hydroxy-Gly or X—NH₂ (X being a peptide or any other compound having a carbonyl group capable of forming a covalent bond with glycine) is accomplished enzymatically in the presence of keto acids, or salts or esters thereof, to provide a good yield without the necessity of catalase or similar enzymatic reaction enhancers. Peptidylglycine α-amidating monooxygenase (PAM) is a preferred enzyme for catalyzing the conversion. Alternatively, peptidylglycine α-hydroxylating monooxygenase (PHM) is utilized to convert X-Gly to X-alpha-hydroxy-Gly which may be recovered, or optionally may be simultaneously or sequentially converted to an amide by either a Lewis base or action of the enzyme peptidyl α-hydroxyglycine α-amidating lyase (PAL). Both PHM and PAL are functional domains of PAM.

The invention belongs to the field of bioengineering and particularly relates to a flavin monooxygenase mutant and a preparation method thereof. The error-prone PCR technology is used to conduct random mutation on wild type flavin monooxygenase to obtain the flavin monooxygenase mutant. The specific enzyme activity of the flavin monooxygenase mutant is improved by 35% compared with that of the wild type flavin monooxygenase. Meanwhile, a yeast expression system is adopted, when the flavin monooxygenase is displayed on the surface of yeasts, yeast cells can serve as enzyme producers and can also serve as carriers in immobilized enzymes, operation of purification, fixation and the like of the enzymes are omitted, the production link is simplified, and the production cost is reduced.

The present invention discloses cytochrome P450BM-3 monoxygenase variant gene and its use. The P450BM-3 variant gene is obtained via error-prone PCR technology mediated random mutant, and has Asp168->Asn, Ala225->Val and Lys440->Asn. The variant gene has nucleotide sequence of SEQ ID No. 1 and encods the cytochrome P450BM-3 monoxygenase. The present invention creates one kind of variant gene P450BM-3 with high catalytic activity, and the variant gene expresses mutant enzyme capable of catalyzing indole to create dye indigo blue in high activity. The present invention provides wide application foreground for the biological conversion of producing dye indigo blue.

The invention belongs to the technical field of bio-catalysis, and in particular relates to a method for improving regeneration of mycobacterium coenzyme and for simultaneously promoting production ofandrostenedione. According to the method, sterol C27 site monooxygenase Cyp125 in a phytosterol metabolic pathway is introduced into steroid transformation microorganism, so as to improve androstenedione conversion efficiency of recombinant microorganism in a fermentation system; and therefore, a novel method for solving a problem of slow metabolism of mycelial phytosterol is offered. Through over-expression of the Cyp125, specific enzyme activity of the mycobacterium sterol C27 site monooxygenase is improved by 22.2%; regeneration of intracellular NAD+ is improved, and an intracellular NAD+/NADH ratio is improved by 131%; and meanwhile, it is unexpectedly discovered that a bacterium biomass amount can be improved by 18.7% through the over-expression of the enzyme, and finally, effects ofshortening a conversion cycle by 24h and improving an AD(D) yield by 10.0% can be achieved.

This invention relates to an isolated polynucleotide encoding a cytochrome P450 monooxygenase that in the presence of linoleic acid results in production of 1-octen-3-ol. The invention also relates to the construction of a recombinant DNA construct comprising all or a portion of the polynucleotide encoding the cytochrome P450 monooxygenases of the invention, in sense or antisense orientation, wherein expression of the recombinant DNA construct results in production of altered levels 1-octen-3-ol in a transformed host cell.

The invention discloses monooxygenase. An amino acid sequence of the monooxygenase is obtained through mutation of an amino acid sequence represented by SEQ ID NO:2. The monooxygenase is used for preparing chiral sulfoxide drugs, has the advantages of mild reaction conditions, environmental friendliness, high yield and product optical purity, few peroxidation products and the like and has very good industrial application prospects in production of a drug proton pump inhibitor for treating gastric ulcer.

Novel genes have been isolated which encode cytochrome P450 and NADPH reductase enzymes of the ω-hydroxylase complex of C. tropicalis 20336. Vectors including these genes, transfected host cells and transformed host cells are provided. Methods of producing of cytochrome P450 and NADPH reductase enzymes are also provided which involve transforming a host cell with a gene encoding these enzymes and culturing the cells. Methods of increasing the production of a dicarboxylic acid and methods of increasing production of the aforementioned enzymes are also provided which involve increasing in the host cell the number of genes encoding these enzymes. A method for discriminating members of a gene family by quantifying the expression of genes is also provided.

The invention provides an immobilized enzyme as well as a preparation method and an application thereof. The immobilized enzyme comprises an enzyme and an amino resin carrier for immobilizing the enzyme, the enzyme is selected from any one of transaminase, ketoreductase, monooxygenase, ammonia lyase, alkene reductase, imine reductase, amino acid dehydrogenase and nitrilase, the amino resin carrieris the cross-linking agent modified amino resin carrier, and a cross-linking agent is a cross-linking agent treated by a high polymer. The amino resin carrier is modified by adopting the cross-linking agent treated by the high polymer, so that enzymes immobilized on the amino resin carrier can form netted cross-linking, the immobilization effect of the enzymes is more stable, and the cyclic utilization efficiency of the enzymes is improved.

The invention relates to the technical field of genetic engineering, and particularly relates to a monooxygenase mutant and a preparation method and application thereof. The monooxygenase mutant is provided with any one of amino acid sequences as shown in formulas (I) and (II), wherein the amino acid sequence in the formula (I) reaches 80% consistency with the amino acid sequence as shown in SEQ ID NO.1; the amino acid sequence in formula (II) is obtained by modifying, substituting, decreasing or increasing one or some amino acid to 23 to 508 amino acid sites of the amino acid sequence as shown in SEQ ID NO.1; 1-34 amino acid is substituted; the mutant has the activity of monooxygenase. According to the method, a plurality of different sites, namely, 23-508 amino acid sites, are mutated; and the result shows that the mutants are capable of improving the activity, stability, soluble expression and selectivity of monooxygenase as well as decreasing the dosage of monooxygenase.

The invention relates to a process for the biocatalytic oxidation of aromatic compounds using recombinant xylene monooxygenase-expressing microorganisms in a biphasic reaction medium.

The invention discloses an Escherichia coli for producing isoeugenol monooxygenase and a construction method and an application of Escherichia coli. The Escherichia coli is named as BL21 (DE3) IEM-PP, and the preservation number is CGMCC No. 8918. The construction method comprises the steps of amplifying whole genome DNA of soil as a template by a PCR (polymerase chain reaction) technology under the action of PCR primers to obtain fragments of which the lengths are 1438bp, carrying out gel extraction and recovery, purifying and linking with a T-vector pET21 (a) to obtain the recombinant strain BL21 (DE3) IEM-PP cloning vector recombinant plasmid PET21 (a)-IEM. The Escherichia coli can be applied in producing vanillin by the biotransformation of isoeugenol via the recombinant strain.