1084 results about "Synzyme" patented technology

Methods, compositions, kits, and apparatus are provided wherein the aminoacyl-tRNA synthetase system is used to analyze amino acids. The method allows very small devices for quantitative or semi-quantitative analysis of the amino acids in samples or in sequential or complete proteolytic digestions. The methods can be readily applied to the detection and/or quantitation of one or more primary amino acids by using cognate aminoacyl-tRNA synthetase and cognate tRNA. The basis of the method is that each of the 20 synthetases and/or a tRNA specific for a different amino acid is separated spatially or differentially labeled. The reactions catalyzed by all 20 synthetases may be monitored simultaneously, or nearly simultaneously, or in parallel. Each separately positioned synthetase or tRNA will signal its cognate amino acid. The synthetase reactions can be monitored using continuous spectroscopic assays. Alternatively, since elongation factor Tu:GTP (EF-Tu:GTP) specifically binds all AA-tRNAs, the aminoacylation reactions catalyzed by the synthetases can be monitored using ligand assays. Microarrays and microsensors for amino acid analysis are provided. Additionally, amino acid analysis devices are integrated with protease digestions to produce miniaturized enzymatic sequenators capable of generating either N- or C-terminal sequence and composition data for a protein or peptide. The possibility of parallel processing of many samples in an automated manner is discussed.

The invention provides methods, systems and reagents for regulating pH-sensitive protein interaction by incorporating non-natural amino acids into the protein (e.g. an antibody, or its functional fragment, derivative, etc.). The invention also relates to specific uses in regulating pH-sensitive binding of antibodies to tumor site, by conferring enhanced tumor-specificity/selectivity. In that embodiment, the non-natural amino acids preferably have desirable side-chain pKa's, such that at below physiological pH (e.g. about pH 6.3-6.5) the non-natural amino acid confer enhanced binding to tumor antigens in acidic environments. Such non-natural amino acids can be incorporated by any suitable means, such as by utilizing a modified aminoacyl-tRNA synthetase to charge the nonstandard amino acid to a modified tRNA, which forms strict Watson-Crick base-pairing with a codon that normally forms wobble base-pairing with natural tRNAs (e.g. the degenerate codon orthogonal system.

The invention provides certain embodiments relating to methods and compositions for incorporating non-natural amino acids into a polypeptide or protein by utilizing a mutant or modified aminoacyl-tRNA synthetase to charge the non-natural amino acid to a the corresponding tRNA. In certain embodiments, the tRNA is also modified such that the complex forms strict Watson-Crick base-pairing with a codon that normally forms wobble base-pairing with unmodified tRNA/aminoacyl-tRNA synthetase pairs.

Thrombopoietic compositions are provided comprising tyrosyl tRNA synthetase polypeptides, including truncations and/or variants thereof. Also provided are methods of using such compositions in the treatment of conditions that benefit from increased thrombopoiesis, such as thrombocytopenia.

The invention provides compounds of the general formula (I)

which are inhibitors of the human aldosterone synthase, and also pharmaceutical compositions containing these compounds, and a method of treating of hyperaldosteronism and/or disorders or diseases that are mediated by 11β-hydroxylase (CYP11B1) with these compounds.

A polypeptide according to the present invention includes: an altered polypeptide obtained by altering an ArgRS, a CysRS, a MetRS, a GlnRS, a GluRS, a LysRS, a TyrRS, or a TrpRS so that an unnatural amino acid is recognized; and an editing polypeptide derived from a PheRS, a LeuRS, an IleRS, a ValRS, an AlaRS, a ProRS, or a ThrRS, the editing polypeptide having been either inserted between a Rossman-fold N domain and a Rossman-fold C domain that exist in the altered polypeptide, or bound to an N terminal of the altered polypeptide. Thus provided are a new aaRS that exhibits high substrate specificity to an unnatural amino acid and a technique that involves the use of such an aaRS.

L-Methionine is produced by culturing a microorganism which is deficient in repressor of L-methionine biosynthesis system and/or enhanced intracellular homoserine transsuccinylase activity is cultured in a medium so that L-methionine should be produced and accumulated in the medium, and collecting the L-methionine from the medium. The microorganism preferably further exhibits reduced intracellular S-adenosylmethionine synthetase activity, L-threonine auxotrophy, enhanced intracellular cystathionine γ-synthase activity and enhanced intracellular aspartokinase-homoserine dehydrogenase II activity. The present invention enables breeding of L-methionine-producing bacteria, and L-methionine production by fermentation.

Provided are histidyl-tRNA synthetase variant polypeptides, X-ray crystallographic and NMR spectroscopy structures of HRS polypeptides, and related compositions and methods for therapy and drug discovery.

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 invention discloses a recombinant microorganism for preparing dharma diene and protopanoxadiol and a construction method of the recombinant microorganism. The construction method of the recombinant bacteria comprises a step of adding dharma diene synthase, protopanoxadiol synthase and nicotinamide adenine dinucleotide phosphate - cytochrome P450 reductase encoding gene into saccharomyces cerevisiae to obtain recombinant bacteria I. According to the recombinant microorganism for preparing the dharma diene and the protopanoxadiol and the construction method of the recombinant microorganism, by means of homologous recombination, the dharma diene synthase, the protopanoxadiol synthase and the nicotinamide adenine dinucleotide phosphate - the cytochrome P450 reductase encoding gene are all added into the saccharomyces cerevisiae to obtain the initial recombinant bacteria, and the effect that the initial recombinant bacteria can produce trace amount of the dharma diene and trace amount of the protopanoxadiol is discovered; tHMG1 activity of the initial recombinant bacteria is further improved, and therefore intermediate recombinant bacteria are obtained, and by means of the intermediate recombinant bacteria, the yield of the dharma diene and the yield of the protopanoxadiol are significantly increased; the activity of one or two or three of ERG1, ERG9 and ERG20 are improved on the basis of the intermediate recombinant bacteria, and the effect that the recombinant bacteria which can be used to increase the yield of the dharma diene and the yield of the protopanoxadiol are constructed is also discovered. By means of the recombinant microorganism for preparing the dharma diene and the protopanoxadiol and the construction method of the recombinant microorganism, the foundation is laid for artificially synthesizing the dammar diene and the protopanoxadiol.

The present invention relates to the isolation and use of an allene oxide synthase enzyme as an antioxidant of lipid peroxides in biological systems. It is based, at least in part, on the discovery that antioxidation is accomplished enzymatically by RPP, a species of allene oxide synthase, in guayule, and on the discovery that the allene oxide synthase RPP disrupts the chain reaction and propagation steps of lipid peroxidation. The present further invention relates to the use of an allene oxide synthase to result in a time-dependent disappearance of conjugated dienes (i.e. lipid hydroperoxides). The allene oxide synthase rapidly converts free or esterified fatty acid peroxides or hydroperoxides into their corresponding epoxides, which, in turn are converted to ketols. The lipid peroxide and hydroperoxide substrates for this enzyme are known to be toxic to biological organisms and can generate additional peroxides by chain propagation reactions. In the presence of an allene oxide synthase these compounds are rapidly and effectively converted to allene oxides (the epoxide), thus breaking the chain reaction.

This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.

Recombinant microorganisms, plants, and plant cells are disclosed that have been engineered to express novel recombinant genes encoding steviol biosynthetic enzymes and UDP-glycosyltransferases (UGTs). Such microorganisms, plants, or plant cells can produce steviol or steviol glycosides, e.g., rubusoside or Rebaudioside A, which can be used as natural sweeteners in food products and dietary supplements.

The subject invention concerns a method of inhibiting an RNA virus infection within a patient by increasing the amount of 2-5 oligoadenylate synthetase (2-5 AS) activity within the patient. Preferably, the preventative and therapeutic methods of the present invention involve administering a nucleotide encoding 2-5 AS, or at least one catalytically active fragment thereof, such as the p40, p69, p100 subunits, to a patient in need thereof. The present inventors have determined that overexpression of 2-5AS causes a reduction in epithelial cell damage, reduction in infiltration of mononuclear cells in the peribronchiolar and perivascular regions, and reduction in thickening of the septa in the lungs. Levels of chemokines, such as MIP1-alpha, are also reduced upon overexpression of 2-5AS. The subject invention also pertains to pharmaceutical compositions containing a nucleotide sequence encoding 2-5 AS and a pharmaceutically acceptable carrier, as well as vectors for delivery of the 2-5 AS nucleotide sequence.

The present invention provides porcine Forssman synthetase (FSM synthase) (Globoside α-N-acetylgalactosaminyltransferase) protein, cDNA, and genomic DNA sequence. Furthermore, the present invention includes porcine animals, tissue and organs as well as cells and cell lines derived from such animals, tissue and organs, which lack expression of functional FSM synthetase. Such animals, tissues, organs and cells can be used in research and in medical therapy, including in xenotransplantation. In addition, methods are provided to prepare organs, tissues, and cells lacking the porcine FSM synthetase gene for use in xenotransplantation.

The invention discloses a novel application of malt oligosaccharide based mycose synthetase and malt oligosaccharide based mycose hydrolase in mycose production. The malt oligosaccharide based mycose synthetase has an amino acid sequence as shown in SEQ ID NO:1; and the malt oligosaccharide based mycose hydrolase has the amino acid sequence as shown in SEQ ID NO:3. The malt oligosaccharide based mycose synthetase and malt oligosaccharide based mycose hydrolase disclosed in the invention also have higher most-suitable reaction temperature and thermal stabilities, and lower most-suitable pH values, so that contamination risks are lowered, production stability is improved, the mycose production efficiency by acting reducing glucidtemns through the malt oligosaccharide based mycose synthetase and malt oligosaccharide based mycose hydrolase and pullulanase in a combined manner is improved and the mycose cost is lowered, therefore, a solid foundation is laid up for the extensive application of the mycose in medicines, foods and cosmetics.

The invention provides a method for preparing trehalose. The method comprises the following step: with starch as a substrate and producing trehalose by using malt oligosaccharyl trehalose synthase and malt oligosaccharyl trehalose hydrolase. According to the method, the trehalose production efficiency is improved by improving processes of liquifying, saccharifying and the like.

The invention provides an escherichia coli engineering bacterium for producing 2'-fucosyllactose. Through a CRISPR/Cas9 gene editing system, related genes of an original strain 2'-fucosyllactose in ananabolic pathway can be knocked out; and a modular metabolic pathway is constructed, the expression level of phosphomannomutase (ManB), mannose-1-phosphateguanylyltransferase (ManC), GDP-mannose-6-dehydrogenase (Gmd), GDP-fucose-synthase (WcaG), L-fucose 1-kinase/GDP-fucose pyrophosphorylase (Fkp) and 2'-fucosyllactose synthetase (FucT2) in the metabolic pathway can be regulated and controlled through different plasmid combinations, so that 2'-fucosyllactose with higher concentration can be accumulated in cells. A method for efficiently producing the 2'-fucosyllactose is also provided.

The invention belongs to the technical field of enzyme engineering, and concretely relates to a pyrimidine nucleoside high-yielding strain and a carbamyl phosphate synthetase adjusting site thereof. The invention provides a pyrimidine nucleoside production Bacillus subtilis mutant strain and a carbamyl phosphate synthetase encoding gene. A key regulation site related to carbamyl phosphate synthetase end product feedback inhibition is defined, and provides reference for breeding of later pyrimidine nucleoside high-yielding strains. The Bacillus subtilis mutant strain allows the output of fermentation process produced nucleoside pyrimidine uridine to reach 14-16g/L, and has substantially improved tolerance to different pyrimidine nucleoside structure analogs.

This invention relates to a new clone mycose synthesized zyme gene and its expression in the correlated host which zyme gene clone belong to thermobifida turning the malt sager to mycose under normal biochemical reaction, its zyme substrate conversion efficiency increases along with the decreasing of the reaction temperature, taking the malt sager or amylum as the raw material and turn it to alpha 1-mycose alpha-1 by cloned.

The invention discloses trehalose synthase of streptomyces griseochromogenes and a coding gene and application of the trehalose synthase. The trehalose synthase gene is cloned from the streptomyces griseochromogenes. The trehalose synthase can convert maltose to generate trehalose, the proper reaction temperature of the trehalose synthase is 15-35 DEG C and is 20-25 DEG C preferably, and the proper pH (potential of hydrogen) of the trehalose synthase ranges from 6.0 to 8.0 and ranges from 7.0 to 7.5 preferably. The conversion efficiency of the trehalose synthase is high, the maltose is used as a substrate, conversion rate of the maltose is about 80% at the reaction temperature of 25 DEG C, and less than 5% of glucose is generated to be beneficial to increase of the conversion rate of the substrate. The trehalose synthase can be used for converting commercial ultrahigh malt syrup to prepare the trehalose, and a novel method using an enzymic method to produce the trehalose industrially can be provided.