42 results about "Acyl Coenzyme A Synthetases" patented technology

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.

The invention discloses a kit and method for clinically detecting serum or plasma free fatty acid. The kit is composed of a reagent 1 and a reagent 2, wherein the reagent 1 comprises a buffer, ATP, a coenzyme A, 4-amino antipyrine, an ion activator, an acyl coenzyme A synthetase, stabilizer, a preservative, a surfactant, and concentrated hydrochloric acid, and the reagent 2 comprises a buffer, a chromogen, a composite sulfhydryl reagent, a peroxidase, an acyl coenzyme A oxidase, a stabilizer, a preservative, a surfactant and concentrated hydrochloric acid. The kit has the advantages of good stability, high specificity of the free fatty acid, and wide linear range. The serum or plasma free fatty acid is detected by the enzyme method, and the method has the advantages of high precision, andsmall measuring error.

Methods for producing polyhydroxyalkanoates (PHAs) from fatty acid biosynthetic pathways using a 3-hydroxy acyl ACP thioesterase, a PHA synthase, and an acyl CoA synthetase, have been developed. Methodology for enabling PHA production from fatty acid biosynthetic pathways in non-native bacterial PHA producers and plants using an enzyme having the catalytic activity of 3-hydroxy acyl ACP thioesterase, an acyl CoA synthetase with substrate specificity for medium chain length 3-hydroxy fatty acids, and a medium chain length PHA synthase, has been developed. Acyl CoA synthetase activity can be supplied either by the endogenous acyl CoA synthetase of the host organism, when sufficiently expressed, or the host organism's activity can be supplemented by the expression of a recombinant acyl CoA synthetase gene. New strategies are described for plant based PHA production in the chloroplasts, cytosol, and peroxisomes of biomass crops as well as the plastids, cytosol, and peroxisomes of oil seed crops.

The invention discloses acyl-coenzyme A synthetase and an application thereof. The amino acid sequence of the acyl-coenzyme A synthetase is as shown in SEQ ID No.1. Further, genetically engineered bacteria capable of secreting the acyl-coenzyme A synthetase can be established, and the acyl-coenzyme A synthetase can be produced by fermentation of recombinant bacteria. The acyl-coenzyme A synthetase disclosed by the invention is the first enzyme which has been ever found to have an activity on trans-8-methyl-6-nonenoic acid or 8-metyl nonanoic acid, and provides a direct substrate for cloning capsaicinoid synthetase. The acyl-coenzyme A synthetase disclosed by the invention has an activity on medium-chain fatty acid and can also be applied to the field of biodiesel.

The invention relates to a reagent for quantitatively detecting the content of free fatty acid in human serum. The reagent comprises a reagent I and a reagent II which are respectively placed, wherein the reagent I comprises sodium dihydrogen phosphate, disodium hydrogen phosphate, magnesium chloride, fatty acyl-coenzyme A synthetase, ascorbic acid oxidase, coenzyme A, ATP and 4-aminoantipyrine, and the reagent II contains sodium dihydrogen phosphate, disodium hydrogen phosphate, phenoxy ethanol, fatty acyl-coenzyme A oxidase, peroxidase, chromogen and a surfactant. The kit and the detection method only need dozens of microlitres of serum without centrifugal or electrophoresis and other separation treatments, are simple and convenient to operate, can be used for meeting the requirement of full-automatic analysis, and are suitable for timely accurate detection of large-scale samples.

The invention relates to a method for synthesizing biodiesel with microorganisms in vivo, which is realized in escherichia coli. The method comprises the following steps of: respectively cloning an acinetobacter wax fat synthetase gene atfA, an escherichia coli acyl coenzyme A synthetase gene fadD, a zymomonas mobilis pyruvate decarboxylase gene pdc and an alcohol dehydrogenase gene adhB gene on a pET expression vector in series to obtain a plasmid pXT11; transferring the plasmid pXT11 into escherichia coli containing a plasmid pMSD8 and a plasmid pMSD15 to obtain an escherichia coli stain XT31; and generating biodiesel by the escherichia coli stain XT31. The method realizes the microbial transformation from sugar to the biodiesel, lays the foundation for biologically preparing the biodiesel from lignocelluloses, and radically solves the problem of insufficient raw material supply in biodiesel preparation. A step of esterifying methanol is not needed in the technology, thereby the cost is greatly reduced and the problems that glycerol as a side product is formed, and the like are avoided.

The present invention relates to a cell for producing acyl glycinates wherein the cell is genetically modified to comprise

• • at least a first genetic mutation that increases the expression relative to the wild type cell of an amino acid-N-acyl-transferase,
  • at least a second genetic mutation that increases the expression relative to the wild type cell of an acyl-CoA synthetase, and
  • at least a third genetic mutation that decreases the expression relative to the wild type cell of at least one enzyme selected from the group consisting of an enzyme of the glycine cleavage system, glycine hydroxymethyltransferase (GlyA) and threonine aldolase (LtaE).

Cells and methods for producing polyhydroxyalkanoates. The cells comprise one or more recombinant genes selected from an R-specific enoyl-CoA hydratase gene, a PHA polymerase gene, a thioesterase gene, and an acyl-CoA-synthetase gene. The cells further have one or more genes functionally deleted. The functionally deleted genes include such genes as an enoyl-CoA hydratase gene, a 3-hydroxyacyl-CoA dehydrogenase, and a 3-ketoacyl-CoA thiolase gene. The recombinant cells are capable of using producing polyhydroxyalkanoates with a high proportion of monomers having the same carbon length from non-lipid substrates, such as carbohydrates.

Described are hosts for overproducing a fatty acid product such as a fatty acid. The hosts include an exogenous nucleic acid encoding a thioesterase and, optionally, an exogenous nucleic acid encoding an acetyl-CoA carboxylase, wherein an acyl-CoA synthetase in the hosts are functionally deleted. The hosts preferably include the nucleic acid encoding the thioesterase at an intermediate copy number. The hosts are preferably recombinantly stable and growth-competent at 37° C. Methods of producing a fatty acid product comprising culturing such hosts at 37° C. are also described.

The invention discloses a method for increasing fermentation yield of polyketone compounds. The method includes steps of: (1) culturing genetically engineered bacteria of an overexpressed acyl-coenzyme A synthesizing enzyme and an acyl carrier protein in a culture medium added with natural grease; and (2) extracting the polyketone compounds from a fermented product. The genetically engineered bacteria are a recombination expression transformant of the overexpressed acyl-coenzyme A synthesizing enzyme and the acyl carrier protein, which is prepared through conjugational transfer by an actinomycete onto recombinant escherichia coli containing a recombinant expression vector expressing the overexpressed acyl-coenzyme A synthesizing enzyme and the acyl carrier protein. In the method, an exogenetic acyl-coenzyme A synthesizing enzyme and an exogenetic acyl carrier protein are overexpressed in the bacteria generating the polyketone compounds through gene engineering technologies, so that by means of cheap natural grease as a raw material, the fermentation yield of the polyketone compounds is increased, wherein the method is never reported in references. The method is simple in operations, can provide precursors for synthesizing the polyketone compounds through the cheap natural grease, is low in cost and is convenient to popularize.

The invention relates to the technical field of free fatty acid determining, in particular to a free fatty acid determining kit. The kit comprises a reagent R1 and a reagent R2, wherein the R1 is prepared from coenzyme A, triphosadenine, acyl coenzyme A synthetase, MgCl2, a Trinder substrate and Tween-20, and has the pH of 7.2; the R2 is prepared from flavin adenine dinucleotide, 4 amino-antipyrine, peroxidase, acyl coenzyme A oxidase, Tween-20, glycerinum, 1 to 1.5g/L of brij35-1 and ethylene diamine tetraacetic acid disodium salt and has the pH of 7.2. By adding different combinations of protein protecting agents, the enzyme activity stability is prolonged; the 37-DEG C stability of the enzyme is prolonged from 7 days to 10 days; the enzyme denaturation is slowed down; the reagent R2 cannot easily generate precipitate.

The invention relates to the field of medical detection technology, and especially relates to a detection kit of non-esterified fatty acids. The kit comprises a reagent 1 and a reagent; the reagent 1 comprises sodium dihydrogen phosphate, coenzyme A, adenosine triphosphate, acyl coenzyme A synthetase, MgCl2, and a Trinder substrate, wherein the pH value is adjusted to 7.2; the reagent 2 comprises sodium dihydrogen phosphate, flavin adenine dinucleotide, 4-aminoantipyrine, acyl coenzyme A synthetase, peroxidase, alkyl glucoside APG0810, mannitol, EDTA-2Na, and polyoxyethylene dodecyl ether, where the pH value is adjusted to 7.2. Surfactants in the reagents are selected, so that activity of various enzymes is stable and are not easy to precipitate, shelf-life is prolonged, enterprise risk cost is reduced, and enterprise benefits are improved.

The present invention relates to genes encoding plant acyl-CoA synthetases and methods of their use. In particular, the present invention is related to plant acyl-coenzyme A synthetases. The present invention encompasses both native and recombinant wild-type forms of the enzymes, as well as mutant and variant forms, some of which possess altered characteristics relative to the wild-type enzyme. The present invention also relates to methods of using acyl-CoA synthetases, including altered expression in transgenic plants and expression in prokaryotes and cell culture systems.

Synergistic pharmaceutical compositions and the methods for preventing and treating proliferative diseases such as cancer and psoriasis. The compositions comprise synergistic combinations of: (i) an acyl-CoA-synthetase enzyme inhibitor (AcsI4), (ii) a compound having an inhibiting effect on enzymes with cyclooxygenase activity (COX); and (iii) a compound selected from a 5-lypoxygenase enzyme (LOX-5) inhibitor and a leukotriene receptor antagonist.

The invention relates to a reagent (kit) for diagnosing/determining amino acid by using an enzyme colorimetric method and an enzyme link method, and also discloses a method for determining the concentration of the amino acid, a composition and components of the reagent, belonging to the technical field of medicine/food/environmental test determination. The reagent (kit) comprises the main components: a buffer solution, a coenzyme, sodium bicarbonate (carbon dioxide), an acetylcoenzyme A, long-chain-acyl-coenzyme A, an amino acid oxidase, a pyruvic oxidase, an acyl-coenzyme A synthetase and a stabilizing agent. A sample and the reagent are mixed according to a certain volume ratio to generate a series of enzymatic reactions, and reactants are placed under an ultraviolet/visible light analyzer for detecting the degree of absorbance rise at the dominant wavelength of 340 nm, thereby measuring and calculating the concentration of the amino acid.