142 results about "Dioxygenase" patented technology

The invention discloses a high-yield delta-tocotrienol genetically engineered bacterium and application thereof, and belongs to the technical field of gene recombination. The invention discloses a genetically engineered bacterium for high yield of delta-tocotrienols, which is characterized in that yarrowia lipolytica ATCC MYA-2613 is used as an original strain, and a ku70 gene is knocked out; the gene is used for expressing and coding 4-hydroxyphenylpyruvate dioxygenase, the gene is used for coding uromelanic acid phytate transferase and the gene is used for coding tocopherol cyclase; intensifying a shikimic acid pathway; a mevalonic acid pathway is strengthened; an isopentenol path is introduced; carrying out protein fusion on the HPD and the HPT; truncating a signal peptide of a gene encoding a tocopherol cyclase and back-filling a leucine defect. The genetically engineered bacterium constructed by the invention has industrial stability, can be amplified by more than 10 times in a 2L fermentation tank level compared with a shake flask level, can produce 616.45 mg / L of delta-tocotrienol after being cultured for 240 hours, and has industrial application prospects.

The invention relates to the technical field of carotenoid cleavage dioxygenase, and discloses a carotenoid cleavage dioxygenase mutant and application thereof. In order to improve the efficiency of converting carotenoid in tobacco into aroma molecule beta-ionone, specific mutation of characteristic sites is carried out on carotenoid cleavage dioxygenase (PhCCD1) from petunia, that is, serine (S) at the 428 site of the carotenoid cleavage dioxygenase is mutated into cysteine (C) or tyrosine (Y) to obtain a mutant S428C or S428Y, and the mutant S428C or S428Y is converted into beta-ionone. The yield of the beta-ionone converted from the carotenoid is greatly improved. Experiments prove that the yields of beta-ionone of S428C and S428Y respectively reach 54.2 mg / L and 50.6 mg / L, which are respectively increased by 69.7% and 58.5% compared with those of a wild strain WT.

The invention relates to the discovery of a biosynthetic gene cluster of an alkaloid, namely, exopoid C, and the biosynthetic gene cluster of the exopoid. A biosynthetic gene cluster exo of an exopoid compound is obtained from a fungus Penicillium sp.HDN14-431 through genome mining, the gene cluster comprises a heme-dependent oxidase gene exoA which is composed of indole dioxygenase and a cytochrome b5 structural domain, a cytochrome P450 enzyme gene exoB and an isopentenyl transferase gene exoC which is composed of indole dioxygenase and a cytochrome b5 structural domain, and the cytochrome P450 enzyme gene exoB and the isopentenyl transferase gene exoC are used as biosynthetic genes of the exopoid compound. All genes in a gene cluster are expressed through an aspergillus nidulans heterologous expression system, two isopentenyl indole alkaloids are obtained in total, and a compound I is a new compound exopoid C. And the compound II, namely the exopoid, is a known compound. The biosynthesis pathway of the compound is reported for the first time.

A Levodopa-selective sensor is provided. A continuous levodopa monitor is also provided. The levodopa sensor includes a sensor area having a working electrode with a levodopa-selective chemistry configured for at least partial implantation in a host. The sensor also includes at least one membrane adjacent the levodopa-selective chemistry. The at least one membrane includes an enzyme domain that at least one enzyme selected from a tyrosinase; a mutated tyrosinase with specificity towards I-3,4-dihydroxyphenylalanine; a dihydroxyphenylalanine 4,5-dioxygenase; a mutated dihydroxyphenylalanine 4,5-dioxygenase with specificity towards I-3,4-dihydroxyphenylalanine; and combinations thereof; a synthase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; a mutated synthase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; a 3,4-dihydroxyphenyl-acetaldehyde synthase; a mutated 3,4- dihydroxyphenyl-acetaldehyde synthase with specificity towards I-3,4-dihydroxyphenylalanine; a dioxygenase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; and a mutated dioxygenase enzyme with specificity towards I-3,4-dihydroxyphenylalanine.

Disclosed herein are inhibitors of 4-hydroxyphenylpyruvate dioxygenase (HPD or HPPD) that have decreased inhibitory activity against sepiapterin reductase (SPR). Disclosed herein are methods of administering an HPD inhibitor alone or in combination with a diet low in tyrosine and / or phenylalanine, with or without dihydrobiopterin (BH2) and / or tetrahydrobiopterin (BH4) supplementation and / or monoamine salvage therapy. The compounds of the present disclosure can be used to treat conditions responsive to modulation or inhibition of HPD, such as inborn errors of metabolism, including tyrosinemia. The compounds of the present disclosure can also be used as herbicides to control weeds.

The invention discloses a preparation method of recombinant catechol-2, 3-dioxygenase and an application of the recombinant catechol-2, 3-dioxygenase. According to the invention, a novel catechol-2, 3-dioxygenase group C23O927 is identified from a constructed metagenome library, and the catechol-2, 3-dioxygenase group C23O927 is a key enzyme for catalyzing degradation of polycyclic aromatic hydrocarbon pollutants. A C23O927-pET32a (+) recombinant expression vector is constructed and converted into host bacteria, heterologous expression of target protein is achieved, and the prepared recombinase belongs to acid enzymes and has good resistance to high temperature, acid, alkali, high-concentration metal ions, organic solvents and surfactants. The recombinase provided by the invention can be used in an acidic environment, has very good enzymatic activity, can catalytically degrade various catechol compounds, can be better used for treating polycyclic aromatic hydrocarbon polluted environments, and has important significance for developing more products which can tolerate various extreme environments and efficiently catalyze degradation of polycyclic aromatic hydrocarbons.

The invention relates to a yarrowia lipolytica strain for producing tyrosol as well as a construction method and application of the yarrowia lipolytica strain, and belongs to the technical field of genetic engineering. According to the present invention, ScADH6, ScARO10F138L and D218G genes are expressed in the yarrowia lipolytica W29 [delta] KU70; the shikimic acid feedback inhibition is relieved by expressing the Y1ARO3K225L gene, the Y1ARO4K2214 gene and the Y1AR07G139S gene; pmLAAD, EcTyrAM53I and A354V genes are introduced, so that the conversion from FAD to FADH2 is promoted, and the supply of NADH (Nicotinamide Adenine Dinucleotide Hormone) in cytoplasm is enhanced; a 4-hydroxyphenylpyruvate dioxygenase (HPD) gene is knocked out; and overexpressing Y1DHS1, Y1DHS2 and Y1DHS3 genes of a shikimic acid pathway to obtain the yarrowia lipolytica strain HY5 for producing tyrosol. The invention lays a foundation for industrial production of tyrosol, and has important social significance and economic value.

The invention discloses a cephalotaxus hainanensis Ch2OGD1 gene and application thereof, and belongs to the technical field of gene engineering. The Ch2OGD1 gene is used for coding a 2-oxoglutaric acid dependent dioxygenase (2OGD) type Pictet-Spengler enzyme, and the Pictet-Spengler enzyme is used for coding a Pictet-Spengler enzyme. Through a tobacco transient expression system and in-vitro enzyme activity determination, the Ch2OGD1 is proved to be capable of specifically catalyzing the condensation of dopamine and 4-hydroxyphenylpropanal to generate 1-phenethyl isoquinoline. The invention provides a directional preparation method of 1-phenethyl isoquinoline based on the Ch2OGD1 gene, provides a key enzyme element for biosynthesis of a cephalotaxine alkaloid precursor compound, and provides a technical basis for expanding the application of the gene in plant genetic engineering. And a potential technical path is provided for relieving the problem of resource shortage of cephalotaxine alkaloids.

A Levodopa-selective sensor is provided. A continuous levodopa monitor is also provided. The levodopa sensor includes a sensor area having a working electrode with a levodopa-selective chemistry configured for at least partial implantation in a host. The sensor also includes at least one membrane adjacent the levodopa-selective chemistry. The at least one membrane includes an enzyme domain that at least one enzyme selected from a tyrosinase; a mutated tyrosinase with specificity towards 1-3,4-dihydroxyphenylalanine; a dihydroxyphenylalanine 4,5-dioxygenase; a mutated dihydroxyphenylalanine 4,5-dioxygenase with specificity towards 1-3,4-dihydroxyphenylalanine; and combinations thereof; a synthase enzyme with specificity towards 1-3,4-dihydroxyphenylalanine; a mutated synthase enzyme with specificity towards 1-3,4-dihydroxyphenylalanine; a 3,4-dihydroxyphenyl-acetaldehyde synthase; a mutated 3,4-dihydroxyphenyl-acetaldehyde synthase with specificity towards 1-3,4-dihydroxyphenylalanine; a dioxygenase enzyme with specificity towards 1-3,4-dihydroxyphenylalanine; and a mutated dioxygenase enzyme with specificity towards 1-3,4-dihydroxyphenylalanine.

A Levodopa-selective sensor is provided. A continuous levodopa monitor is also provided. The levodopa sensor includes a sensor area having a working electrode with a levodopa-selective chemistry configured for at least partial implantation in a host. The sensor also includes at least one membrane adjacent the levodopa-selective chemistry. The at least one membrane includes an enzyme domain that at least one enzyme selected from a tyrosinase; a mutated tyrosinase with specificity towards I-3,4-dihydroxyphenylalanine; a dihydroxyphenylalanine 4,5-dioxygenase; a mutated dihydroxyphenylalanine 4,5-dioxygenase with specificity towards I-3,4-dihydroxyphenylalanine; and combinations thereof; a synthase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; a mutated synthase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; a 3,4-dihydroxyphenyl-acetaldehyde synthase; a mutated 3,4- dihydroxyphenyl-acetaldehyde synthase with specificity towards I-3,4-dihydroxyphenylalanine; a dioxygenase enzyme with specificity towards I-3,4-dihydroxyphenylalanine; and a mutated dioxygenase enzyme with specificity towards I-3,4-dihydroxyphenylalanine.

The invention discloses a tobacco carotenoid dioxygenase NtCCD10 mutant as well as a genetically engineered bacterium and application thereof, and belongs to the technical field of oxidation aroma production of carrots. The thermal stability of the mutant NtCCD10-G189P-K279R disclosed by the invention is obviously improved compared with that of the NtCCD10, and the residual enzyme activity of the NtCCD10-G189P-K279R is 1.25 times that of the NtCCD10 under the condition that the mutant NtCCD10-G189P-K279R is incubated for 30 minutes at the temperature of 50 DEG C. A recombinant strain is constructed based on different carotenoid dioxygenase, and beta-carotene can be cracked to produce an aroma substance beta-ionone; the beta-carotene degradation rate of the constructed recombinant strain reaches 54.59% or above within 24 h, and the yield of beta-ionone reaches 20.721 mg / L or above; after the recombinant strain is sprayed to fermented tobacco shreds, the tobacco shred fragrance can be obviously improved, and the tobacco leaf quality is improved.

The present invention discloses a mutant of a dioxygenase for biocatalytic synthesis of vanillin, a preparation method thereof and applications. The mutant is such that at least one of the 332nd, 401st, and 553rd positions in the amino acid sequence shown in SEQ ID NO.1 is mutated; by mutating the aromatic dioxygenase sequence of 4-vinylguaiacol and through screening, this series of mutants can efficiently catalyze the formation of vanillin from 4-vinylguaiacol, and it has a yield 5 to 7 times higher than that of the template.

Pigment compositions comprising betalain pigment molecules are provided. Compositions comprising betanin and betanidin are also provided as are compositions comprising less than 5% sugars by weight. Cells comprising at least one copy of an exogenous tyrosine hydroxylase gene, at least one copy of an exogenous L-DOPA oxidase gene, at least one copy of an exogenous DOPA 4,5-dioxygenase (DOD) gene, at least one copy of an exogenous NADPH cytochrome P450 reductase gene, and at least one copy of a cyclo- DOPA-5-O-glucosyltransferase gene integrated into the genome of the cell are also provided.

The invention relates to the technical field of genetic engineering, and discloses a genetically engineered bacterium for producing beta-ionone at high yield and application of the genetically engineered bacterium. A synthetic biological plasmid modularization technology is adopted, coding genes of carotenoid cleavage dioxygenase NtCCD1 in tobacco are excavated, and specific soluble protein tags including a thioredoxin tag, a maltose binding protein tag and a glutathione transferase tag are screened, so that the content of the carotenoid cleavage dioxygenase NtCCD1 in the tobacco is determined. The NtCCD1 is subjected to single-point mutation modification of mutation of 431st-site histidine into tyrosine, lysine or glutamine, and the fermentation enzyme and the gene engineering bacterium which can efficiently catalyze splitting decomposition of beta-carotene in tobacco to produce beta-ionone are constructed. The biosynthesis of the tobacco aroma component beta-ionone is realized, and the method has high efficiency.

The present disclosure relates in some aspects to constructs and methods for increasing solid organ transplant survival. More specifically, aspects of the present disclosure relate to methods and constructs for administering full-length PD-L1 and one immune modulators, such as one or more of: CTLA-4, Qa-1d, HLA-E, HLA-G, CMTM6, and Indolamine-2,3-dioxygenase (IDO), by ex vivo perfusion in order to increase solid organ transplant survival.

The invention belongs to the technical field of biology, and particularly relates to a pentachloronitrobenzene rapid detection method based on enzyme catalysis reaction, which comprises the following steps: converting pentachloronitrobenzene into pentachlorophenol and nitrate radicals through an enzyme system consisting of iron thioflavin protein reductase, ferredoxin and nitrobenzene dioxygenase; and then nitrate radicals are specifically detected by a Griess reagent method so as to indirectly reflect quintozene. The detection method disclosed by the invention is good in specificity, short in detection time and remarkable in visual detection effect, and can be used for on-site rapid detection of quintozene.

Disclosed are proteolysis-targeting chimeric molecules (PROTACs) that induce degradation of IDO protein. The disclosed PROTACs typically include a first targeting moiety that binds to IDO (Miro). The first targeting moiety typically is linked via a bond or a linker (L) to a second targeting moiety that binds to an E3 ubiquitin ligase (ME3). As such, the disclosed PROTACS may be described as having a formula MIDO-L-ME3 or ME3-L-MIDO.

The invention relates to the field of tobacco, in particular to an expression cassette, an expression vector and a preparation for improving tobacco quality and application thereof. The present invention provides an expression cassette comprising: a gene encoding carotenoid cleavage dioxygenase and a promoter. Probiotic escherichia coli E. coli Nissle 1917 is used as an expression host, genetically engineered bacteria for heterologously expressing carotenoid cleavage dioxygenase are constructed, endotoxin is prevented from being introduced in the tobacco leaf fermentation process, and a biological preparation capable of increasing aroma substances of tobacco leaves can be obtained through simple cell disruption. Meanwhile, the content of aroma substances such as geranyl acetone and ionone in the tobacco leaves can be remarkably increased through tobacco leaf fermentation.

The present invention provides a tricyclic diterpenoid compound having anti-influenza virus activity. The tricyclic diterpenoid compound not only has better anti-H1N1 virus activity, but also is low in toxicity. The invention also discloses a preparation method of the tricyclic diterpenoid compound with antiviral activity. The preparation method comprises the following steps: (1) obtaining a fermentation product rich in the tricyclic diterpenoid compound through microbial fermentation culture; a recombinant vector adopted in the microbial fermentation culture carries nucleotide sequences: PQ793156 (dioxygenase gene), PQ793157 (FAD dependent monooxygenase gene), PQ793158 (terpenoid synthase gene), PQ793159 (UbiA family isopentenyl transferase gene), PQ793160 (GGPP synthase gene) and PQ793161 (amino transferase gene). And (2) separating and purifying the fermentation product obtained in the step (1) to obtain the tricyclic diterpenoid compound.

Disclosed herein is a molecule, or a pharmaceutically acceptable salt thereof, that has a formula M1DO1-L-ME3, M1DO1 is a moiety that binds to IDO1, L is a linker covalently attaching M1DO1 and ME3, and ME3 is a moiety that binds to an E3 ubiquitin ligase. Disclosed herein are also the uses of the molecule, or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the same, in a method of treating cancer in a subject in need thereof.

Various methods and compositions of treating 4-hydroxyphenylpyruvate dioxygenase-like (HPDL)-related diseases or disorders are presented herein. Also presented herein are methods of increasing CoQ10 biosynthesis, and methods of determining whether a subject will benefit from a CoQ10 or CoQ10 alternative treatment. Also presented herein are pharmaceutical compositions and dosage forms comprising 4-hydroxymandelic acid (4-HMA), and / or its metabolites. Further presented herein are compounds that inhibit 4-hydroxyphenylpyruvate dioxygenase-like (HPDL). Further presented herein are methods of identifying and / or assessing modulators of HPDL. Yet further presented herein are example methods and systems for isotopic labelling in cells by metabolizing cells in the presence of gaseous isotopic tracer.

Disclosed herein are compositions comprising (a) a pyridine carboxylate herbicide or an agriculturally acceptable N-oxide, salt, or ester thereof and (b) an HPPD inhibitor herbicide or an agriculturally acceptable salt or ester thereof. Also disclosed herein are methods of controlling undesirable vegetation, comprising applying to vegetation or an area adjacent the vegetation or applying in soil or water to control the emergence or growth of vegetation (a) a pyridine carboxylate herbicide or an agriculturally acceptable N-oxide, salt, or ester thereof and (b) an HPPD inhibitor herbicide or an agriculturally acceptable salt or ester thereof.

The present application belongs to the technical field of quorum quenching enzyme preparation, and particularly relates to a multi-target quorum quenching enzyme preparation and a preparation method and application thereof, such as being mainly used for inhibiting the virulence factors of Pseudomonas aeruginosa, and including acyl homoserine lactone acyltransferase AiiO protein and 3-hydroxy-4-oxoquinolone 2,4-dioxygenase AqdC protein, and the mass ratio of the two is 20:(1-25); such as being mainly used for inhibiting the virulence factors and the synthesis of biological membranes of Pseudomonas aeruginosa, and further including deferiprone, and the mass ratio of AiiO, AqdC and deferiprone is 20:(1-25):(0.695-2.78), the present application can target the Las, Rhl and PQS of Pseudomonas aeruginosa, the yield and stability of the prepared AqdC protein are both high, the two types of enzymes are used in combination after the regulation amount, and the virulence factors regulated by different quorum sensing pathways in Pseudomonas aeruginosa are effectively inhibited; the regulation amount of deferiprone is used in combination with the two types of quorum quenching enzymes, and the virulence and biological membranes of Pseudomonas aeruginosa can be effectively inhibited in a comprehensive manner.

Indoleamine 2,3-dioxygenase (IDO) is a heme-binding enzyme involved in pathophysiological processes including, but not limited to, antimicrobial and antitumor defense, neuropathology, immunoregulation, and antioxidant activity. Aspects of the disclosure relate to variants of IDO that comprise one or more amino acid variations relative to a wild-type IDO and / or that are connected to a synthetic polymer, such as poly (ethylene glycol). Embodiments of the disclosure provide variant IDOs that have improved properties, such as resistance to oxidative conditions, protein solubility, lower propensity to aggregate in vivo, increased half-life in the circulatory system upon administration to a subject, increased binding to a cell or tissue, or any combination thereof. Accordingly, IDO variants of the disclosure can be administered to a subject in need thereof, such as a subject having, suspected of having, or at risk of developing a disease, disorder, or condition, such as one associated with inflammation.

The invention provides 2-oxoglutaric acid dependent dioxygenase 2-ODD of glycyrrhiza glabra as well as a coding gene and application of the 2-oxoglutaric acid dependent dioxygenase 2-ODD. The coding gene of the 2-oxoglutaric acid dependent dioxygenase 2-ODD of the glycyrrhiza glabra, which is provided by the invention, has a nucleotide sequence as shown in SEQ ID NO. 1. The protein coded by the Gg2-ODD can be used for catalytically synthesizing glabridin or a precursor compound thereof. The synthesis method disclosed by the invention is economical, efficient, green and environment-friendly, and the synthesized glabridin or the precursor thereof is high in purity. The enzyme participates in synthesis of glabridin, provides an important element for synthetic organisms of glabridin, and also provides a basis for discussing a regulation mechanism of glabridin synthesis in the future.

The present invention relates to processes for the formation of pyridinedicarboxylic acid (PDCA), in particular, 2,4-pyridinedicarboxylic acid (2,4-PDCA) and 2,5-pyridinedicarboxylic acid (2,5-PDCA), and mono- and diester derivatives thereof, from 3,4-dihydroxybenzoic acid, via a biocatalytic reaction using, for example, a protocatechuate dioxygenase such as protocatechuate 4,5-dioxygenase or protocatechuate 2,3-dioxygenase, and a nitrogen source. The invention also relates to copolymers that comprise the pyridinedicarboxylic acid monomers and derivatives thereof, processes for the formation of the copolymers and uses for the copolymers.

The invention discloses an enzyme activity detection reagent for plant alpha-ketoglutaric acid / Fe (II) dependent dioxygenase, a kit and application, and belongs to the technical field of biology. The enzyme activity detection reagent comprises a reagent I and a reagent II, wherein the reagent I comprises the following components: 1-5 mM of dithiothreitol, 0.1-1 mM of oxoglutaric acid, 0.1-1 mM of sodium ascorbate and 0.01-0.1 mg / mL of catalase, and sterilized distilled water is used as a solvent; the reagent II is prepared from 2 to 3 mg of FeSO4. 7H2O solid. Reagents for enzyme catalytic reaction are mixed according to a specific proportion to prepare the reagent I and the reagent II, the reagent I and the reagent II are directly subjected to one-time mixed reaction with a sample to be detected and a substrate during use, the operation is simple, convenient and rapid, the problems of errors, omission and the like caused by mixing reaction components for multiple times are avoided, and the detection accuracy is improved. And the method can also be used for detecting the enzyme activity of alpha-ketoglutaric acid / Fe (II) dependent dioxygenase of various plants, and has universality.

Provided are methods for treating cancer by administrating to an individual who has the cancer an inhibitor of 4-hydroxyphenylpyruvate dioxygenase (HPPD) and an immune checkpoint inhibitor. Also provided are methods for promoting translation of proteins within cells such that proteins include one or more misincorporated amino acids to thereby develop new proteins that contain neoantigens. Vaccines that include the neoantigens are included. Methods for producing and detecting misincorporation of a tyrosine in proteins expressed by cells are also provided.

This disclosure provides a method for treating a cancer patient comprising administering to the patient a therapeutically effective amount of an anti-PD-1 antagonist, for example, an anti-PD-1 or anti-PD-L1 antibody, in combination with an indolamine 2,3-dioxygenase inhibitor, wherein the patient is identified as exhibiting a combined biomarker comprising (a) a high IFNγ inflammatory signature score and (b) a low tryptophan 2,3-dioxygenase 2 (TDO2) gene expression score. The high IFNγ inflammatory signature score is determined by measuring the expression of a panel of IFNγ-related inflammatory genes in a cancer sample obtained from the patient, wherein the gene panel comprises, for example, IFNγ, CXCL10, CXCL9, HLA-DRA, IDO1, and STAT1. In some respects, the gene panel also includes CCR5, CXCL11, GZMA, and PRF1.In some aspects, the genetic panel comprises CXCR6, TIGIT, PD-L1, PD-L2, LAG3, NKG7, PSMB10, CMKLR1, CD8A, IDO1, CCL5, CXCL9, HLA.DQA1, CD276, HLA.DRB1, STAT1, HLA.E and TDO2.

The invention relates to a strain for degrading beta-carotene, a crude enzyme solution, dioxygenase and application, and relates to the technical field of genetic engineering and microbial fermentation. The invention relates to a strain for degrading beta-carotene, the preservation name of the strain is Escherichia coli LZU-E1, and the strain is preserved in the China General Microbiological Culture Collection Center (CGMCC) with the preservation number of CGMCC No.34862. The invention further discloses a preparation method of the strain for degrading beta-carotene. The method comprises the following steps: screening a strain for efficiently degrading beta-carotene, extracting a crude enzyme solution, researching the enzymatic property of the crude enzyme solution, and mining a new dioxygenase gene for degrading beta-carotene from genome information of the strain by utilizing a molecular biology means, so that the isoprene-reduced natural perfume is efficiently obtained.