167 results about "Acyltransferases" patented technology

A method for producing a polyhydroxyalkanoate with improved productivity and composition is provided. Polyhydroxyalkanoate is produced by a bacterium for producing polyhydroxyalkanoate in which a gene encoding acetyl-CoA acyltransferase is disrupted.

The present invention is directed to a process to remove impurities from triacylglycerol oil including mixing the oil and a fluidic agent, pumping the mixture through a flow-through hydrodynamic cavitation apparatus at a pre-determined inlet pump pressure, creating hydrodynamic cavitation in the mixture, maintaining the hydrodynamic cavitation for a pre-determined period of time, moving the impurities from the oil to the fluidic agent, and then separating the fluidic agent from the oil. The impurities can include phytosterols, sterol glucosides, acylated sterol glucosides, in which case the fluidic agent is water, an alkali hydroxide, an inorganic base, an organic base, phosphoric acid, citric acid, acetic acid or a mixture thereof. The impurities may also include phosphatides, in which case and the fluidic agent comprises water and an enzyme such as phospholipase, a lipid acyltransferase or a mixture thereof.

The present invention relates to a process for the production of long-chain polyunsaturated fatty acids in an organism by introducing, into the organism, nucleic acids which code for polypeptides with acyl transferase activity. These nucleic acid sequences, if appropriate together with further nucleic acid sequences which code for polypeptides of the fatty acid or lipid metabolism biosynthesis, can advantageously be expressed in the organism. Furthermore, the invention relates to a method for the production of oils and / or triacylglycerides with an elevated content of long-chain polyunsaturated fatty acids.The invention furthermore relates to the nucleic acid sequences, nucleic acid constructs, vectors and organisms comprising the nucleic acid sequences according to the invention, vectors comprising the nucleic acid sequences and / or the nucleic acid constructs and to transgenic organisms comprising the abovementioned nucleic acid sequences, nucleic acid constructs and / or vectors.A further part of the invention relates to oils, lipids and / or fatty acids produced by the process according to the invention and to their use.

The present invention relates to a method for the production of a variant lipid acyltransferase comprising the steps of: (i) selecting a parent enzyme which is a lipid acyltranserase enzyme; (ii) modifying one or more amino acids to produce a variant lipid acyltransferase; (iii) testing the activity of the variant lipid acyltransferase on a galactolipid and / or phospholipid and / or triglyceride substrate; (iv) selecting a variant enzyme with enhanced activity towards galactolipids compared with the parent enzyme; (v) providing a Bacillus licheniformis cell; (vi) transforming the Bacillus licheniformis cell with a heterologous nucleotide sequence encoding said variant lipid acyltransferase; and (iii) expressing said variant lipid acyltransferase in the cell under the control of a promoter sequence. The variant lipid acyltransferase can undergo post-translations modification, truncation and / or clipping, i.e., to remove a signal peptide. In addition, the present invention further relates to the use of Bacillus licheniformis to express a lipid acyltransferase, a Bacillus licheniformis host cell comprising a heterologous lipid acyltransferase and a vector comprising a nucleotide sequence encoding a lipid acyltransferase operably linked to a promoter sequence homologous to B. licheniformis.

The present invention relates to a method of reducing and / or removing diglyceride from an edible oil, comprising a) admixing an edible oil with an acyl acceptor substrate and a diglyceride:glycerol acyltransferase, wherein the diglyceride:glycerol acyltransferase is characterized as an enzyme which in an edible oil is capable of transferring an acyl group from a diglyceride to glycerol. Preferably, the diglyceride:glycerol acyltransferase comprises the amino acid sequence motif GDSX, wherein X is one or more of the following amino acid residues L, A, V, I, F, Y, H, Q, T, N, M or S. Furthermore the present invention relates to the use of a diglyceride:glycerol acyltransferase characterized as an enzyme which in an edible oil is capable of transferring an acyl group from a diglyceride to glycerol, in the manufacture of an edible oil, for reducing and / or removing (preferably selectively reducing and / or removing) diglyceride from said edible oil and to the use of said enzyme in the manufacture of a foodstuff comprising an edible oil for improving the crystallization properties of said foodstuff.

Nucleic acid compositions encoding polypeptide products with diglyceride acyltransferase and / or monoacylglycerol acyltransferase activity, as well as the polypeptide products encoded thereby, i.e., mammalian DGAT2α, MGAT1, or MGAT2 polypeptide products, and methods for producing the same, are provided. Also provided are: methods and compositions for modulating DGAT2α, MGAT1, or MGAT2 activity; DGAT2α, MGAT1, or MGAT2 transgenic cells, animals and plants, as well as methods for their preparation; and methods for making diglyceride, diglyceride compositions, triglycerides and triglyceride compositions, as well as the compositions produced by these methods. The subject methods and compositions find use in a variety of different applications, including research, medicine, agriculture and industry applications.

This disclosure describes genetically modified photosynthetic microorganisms, e.g., Cyanobacteria, that contain one or more exogenous genes encoding a phospholipase and / or thioesterase, which are capable of producing an increased amount of lipids and / or fatty acids. This disclosure also describes genetically modified photosynthetic microorganisms that contain one or more exogenous genes encoding a diacyglycerol acyltransferase, a phosphatidate phosphatase, and / or an acetyl-CoA carboxylase, which are capable of producing increased amounts of fatty acids and / or synthesizing triglycerides, as well as photosynthetic microorganism comprising mutations or deletions in a glycogen biosynthesis or storage pathway, which accumulate a reduced amount of glycogen under reduced nitrogen conditions as compared to a wild type photosynthetic microorganism.

A method for producing a polyhydroxyalkanoate with improved productivity and composition is provided. Polyhydroxyalkanoate is produced by a bacterium for producing polyhydroxyalkanoate in which a gene encoding acetyl-CoA acyltransferase is disrupted.

A method for reducing the likelihood of developing non-alcoholic steatohepatitis (NASH) in an individual diagnosed with non-alcoholic fatty liver disease involves providing in the gut of an individual a population of beneficial bacteria selected from the group consisting of Lactobacillus species, and at least 6 grams per day of fiber to the individual to maintain a therapeutically effective amount of the beneficial bacteria in the gut of the individual. In certain embodiments, monoacylglycerolacyltransferase-3 (MGAT3) synthesis is inhibited to lower triacylglycerol (TAG) production, while in others, expression of diacylglycerolacyltransferase-2 (DGAT-2) is inhibited. The beneficial bacteria are preferably modified to produce increased amounts of butyrate and are also encapsulated in a frangible enclosure. Levels of Roseburia are preferably increased while the levels of Akkermansia spp. in the individual's gut microbiome are reduced.

Disclosed are novel acyl-CoA synthetases and novel acyltransferases, nucleic acid molecules encoding the same, recombinant nucleic acid molecules and recombinant host cells comprising such nucleic acid molecules, genetically modified organisms (microorganisms and plants) comprising the same, and methods of making and using the same. Also disclosed are genetically modified organisms (e.g., plants, microorganisms) that have been genetically modified to express a PKS-like system for the production of PUFAs (a PUFA PKS system or PUFA synthase), wherein the organisms have been modified to express an acyl-CoA synthetase, to express an acyl transferase, to delete or inactivate a fatty acid synthase (FAS) expressed by the organism, to reduce competition for malonyl CoA with the PUFA synthase or toincrease the level of malonyl CoA in the organism, and in one aspect, to inhibit KASII or KASIII. Additional modifications, and methods to make and use such organisms, in addition to PUFAs and oils o btained from such organisms, are disclosed, alone with various products including such PUFAs and oils.

Nucleic acid compositions encoding polypeptide products with diglyceride acyltransferase and / or monoacylglycerol acyltransferase activity, as well as the polypeptide products encoded thereby, i.e., mammalian DGAT2α and MGAT1 polypeptide products, and methods for producing the same, are provided. Also provided are: methods and compositions for modulating DGAT2α and MGAT1 activity; DGAT2α and MGAT1 transgenic cells, animals and plants, as well as methods for their preparation; and methods for making diglyceride, diglyceride compositions, triglycerides and triglyceride compositions, as well as the compositions produced by these methods. The subject methods and compositions find use in a variety of different applications, including research, medicine, agriculture and industry applications.

This disclosure describes genetically modified photosynthetic microorganisms, including Cyanobacteria, that contain one or more mutations or deletions in a glycogen biosynthesis or storage pathway, which accumulate a reduced amount of glycogen as compared to a wild type Cyanobacterium, and which are capable of producing an increased amount of lipids and / or fatty acids. In certain embodiments, the genetically modified photosynthetic microorganisms also contain one or more exogenous genes encoding a diacyglycerol acyltransferase, a phosphatidate phosphatase, and / or an acetyl-CoA carboxylase, and which are capable of producing increased amounts of lipids or fatty acids and / or synthesizing triglycerides.

The invention relates to a production method for synthesizing C8:0 / C10:0 / C12:0 / C14:0 medium-chain fatty acid and ethyl ester thereof by using engineering Escherichia coli. The production method comprises the following steps of: cloning thioesterase genes with different specificities chFatB / ucFatB / ccFatB in cuphea / laurel / camphorwood into an expression vector pETDuet-1 to obtain a corresponding recombinant plasmid; transferring the recombinant plasmid into Escherichia coli BL21, performing isopropyl-beta-d-thiogalactoside (IPDT) induction expression, and specifically hydrolyzing acyl carrier protein (ACP) with specific chain length by using the expressed thioesterase to obtain medium-chain fatty acid; cloning acyltransferase gene (atfA) in acinetobacter into expression vectors pETDuet-chFatB / pETDuet-ucFatB / pETDuet-ccFatB to obtain corresponding recombinant plasmids; and transferring the recombinant plasmids into the Escherichia coli BL21, performing IPTG induction expression, performing acyl transfer reaction on exogenously fed ethanol and acyl-CoA formed by the medium-chain fatty acid intracellularly by using the co-expressed acyltransferase to obtain fatty acid ethyl ester.

A process of water degumming an edible oil (preferably a crude edible oil) comprising the steps of: a) admixing approximately 0.1-5% w / w water with an edible oil (preferably a crude edible oil) and a lipid acyltransferase, b) agitating the admixture for between about 10 minutes and 180 minutes at about 45 to about 900 C, and c) separating the oil phase and the gum phase. Preferably said lipid acyltransferase is a polypeptide having lipid acyltransferase activity which polypeptide is obtained by expression of the nucleotide sequence shown as SEQ ID No. 49 or a nucleotide sequence which as has 70% or more identity therewith; and / or is obtained by expression of a nucleic acid which hybridises under medium stringency conditions to a nucleic probe comprising the nucleotide sequence shown as SEQ No. 49; and / or is a polypeptide having lipid acyltransferase activity which polypeptide comprises the amino acid sequence shown as SEQ ID No. 68 or an amino acid sequence which as has 70% or more identity therewith. In one embodiment the lipid acyltransferase is preferably used in combination with a phospholipase C enzyme. A process for modifying the gum phase of a degummed oil using a lipid acyltransferase is also taught herein.

The present invention is directed to a process to remove impurities from triacylglycerol oil including mixing the oil and a fluidic agent, pumping the mixture through a flow-through hydrodynamic cavitation apparatus at a pre-determined inlet pump pressure, creating hydrodynamic cavitation in the mixture, maintaining the hydrodynamic cavitation for a pre-determined period of time, moving the impurities from the oil to the fluidic agent, and then separating the fluidic agent from the oil. The impurities can include phytosterols, sterol glucosides, acylated sterol glucosides, in which case the fluidic agent is water, an alkali hydroxide, an inorganic base, an organic base, phosphoric acid, citric acid, acetic acid or a mixture thereof. The impurities may also include phosphatides, in which case and the fluidic agent comprises water and an enzyme such as phospholipase, a lipid acyltransferase or a mixture thereof.

The present invention is directed to lecithin:cholesterol acyltransferase-like polypeptides (LCAT) and acyl CoA:cholesterol acyltransferases-like polypeptides (ACAT). The invention provides polynucleotides encoding such cholesterol:acyltansferases-like polypeptides, polypeptides encoded by such polynucleotides, and the use of such polynucleotides to alter sterol composition and oil production in plants and host cells. Also provided are oils produced by the plants and host cells containing the polynucleotides and food products, nutritional supplements, and pharmaceutical composition containing plants or oils of the present invention. The polynucleotides of the present invention include those derived from plant sources.

This invention relates to plant LPAATs, means to identify such proteins, amino acid and nucleic acid sequences associated with such protein, and methods to obtain, make and / or use such plant LPAATs. Purification, especially the removal of plant membranes and the substantial separation away from other plant proteins, and use of the plant LPAAT is provided, including the use of the protein as a tool in gene isolation for biotechnological applications. In addition, nucleic acid sequences encoding LPAAT protein regions are provided, and uses of such sequences for isolation of LPAAT genes from plants are considered.

The present invention claims an isolated polypeptide having L-amino-acid-N-acyl transferase enzymatic activity and a modified microorganism in which this enzyme is overexpressed. Substrates of said enzyme include mainly methionine and their derivatives or analogs. Overexpression in sulphur-containing amino acid producing microorganisms permits the production of large amounts of N-acylated sulphur-containing amino acids. The isolation of the N-acylated sulphur-containing amino acids from the fermentation medium is also claimed.

The present invention relates to diacylglycerol acyltransferase genes and proteins, and methods of their use. In particular, the invention describes genes and proteins that exhibit both long-chain acyltransferase and acetyltransferase activity. The present invention encompasses both native and recombinant wild-type forms of the transferase, as well as mutants and variant forms, some of which possess altered characteristics relative to the wild-type transferase. The present invention also relates to methods of using diacylglycerol acyltransferase genes and proteins, including in their expression in transgenic organisms and in the production of acetyl-glycerides in plant oils, and in particular seed oils.

A method including advancing a delivery device through a lumen of a blood vessel to a particular region in the blood vessel; and introducing a composition including a sustained-release carrier and an apolipoprotein A-I (apo A-I) synthetic mimetic peptide into a wall of the blood vessel at the particular region or a perivascular site, wherein the peptide has a property that renders the peptide effective in reverse cholesterol transport. A composition including an apolipoprotein A-I (apo A-I) synthetic peptide, or combination of an apo A-I synthetic mimetic peptide and an Acyl CoA cholesterol: acyltransferase (ACAT) inhibitor in a form suitable for delivery into a blood vessel, the peptide including an amino acid sequence in an order reverse to an order of various apo A-I mimetic peptides, or endogenous apo A-I analogs, or a chimera of helix 1 and helix 9 of endogenous apo A-I.

The invention relates to the field of pharmacy, and provides an improved method for preparing amoxicillin by an enzymic method, and a product obtained by the improved method for preparing amoxicillin by the enzymic method. The method comprises the following steps of: 1) dissolving 6-aminopenicillanic acid (6-APA) at the temperature of between 10 and 20 DEG C by using water or / and aqueous solution of ammonia which has the pH value of 7.0 to 8.0, and adding D-p-Hydroxyphenylglycine methyl ester hydrochlorid and penicillin G acyltransferase; 2) adjusting the pH value of a solution obtained in the step 1) to be 6.0 to 6.5, and reacting at the temperature of between 21 and 30 DEG C until the content of 6-APA is less than 5mg / ml to obtain a solution of an amoxicillin product; and 3) separating the penicillin G acyltransferase from the solution of the amoxicillin product, adjusting by using hydrochloric acid until the solution of the amoxicillin product is clarified, adding the aqueous solution of ammonia, adjusting the pH value to be 5.5 to 6.5, and crystallizing at the temperature of between 0 and 5 DEG C to obtain amoxicillin. By the improved method for preparing amoxicillin by the enzymic method, the quality of the amoxicillin product is greatly improved, and the medication safety of the amoxicillin product is further improved.

The invention relates to a one-step enzymatic method for preparing 7-aminocephalosporanic acid. The method for preparing the 7-aminocephalosporanic acid comprises four steps of: 1, treating fermentation liquid of cephalosporin C; 2, performing enzymolysis of the cephalosporin C under the action of acyltransferase of the cephalosporin C to obtain the 7-aminocephalosporanic acid; 3, treating the lysate of 7-aminocephalosporanic acid obtained through the enzymolysis; and 4, crystallizing and drying the 7-aminocephalosporanic acid.

The present invention is related to biosynthetic oil compositions and methods of making thereof. In some embodiments, the invention relates to the use of endogenous enzymes in plants capable of synthesizing oil. In preferred embodiments, said plants are algae. In further embodiments, said algae are from the family Chlamydomonas, Nannochloropsis, Dunaliella, Chlorella and Scenedesmus. In still further embodiments, said endogenous enzymes are diacylglycerol acyltransferases.

The invention relates to a method for preparing 7-ACA (aminocephalosporanic acid) and obtaining alpha-aminoadipic acid. The method comprises the following steps of (1) treating cephalosporin C fermentation liquid; (2) performing an enzymolysis reaction of cephalosporin C under the effect of cephalosporin C acyltransferase to obtain 7-ACA; (3) treating the 7-ACA lysate after the enzymolysis; (4) separating out the 7-ACA; (5) separating alpha-aminoadipic acid from the separated 7-ACA lysate.

PCT No. PCT / EP95 / 02876 Sec. 371 Date Feb. 19, 1997 Sec. 102(e) Date Feb. 19, 1997 PCT Filed Jul. 18, 1995 PCT Pub. No. WO96 / 02663 PCT Pub. Date Feb. 1, 1996An improvement for enzymatically syhnthesizing a beta -lactam compound is presented. The improvement comprises catalyzing the acylation of an amino beta -lactam with an acylating agent for at least 5 hours with an amidase or acylase, wherein the concentration of each reactant is constantly higher than 70% of the lowest value of the saturated concentration of both reactants.

The method for preparing glutaryl-7-aminocephaphylanic acid acylase is characterized by that it utilizes colibacillus gene engineering bacterium to make shake-flask strain culture in culture medium, fermentation strain culture and fermentation culture, then utilizes centrifuge to collect thallus, soakes to extract enzyme liquor, makes the enzyme liquor undergo the processes of plate-frame filtration, ultrafiltration concentration, purification and fixation so as to obtain the invented product GL-7-ACA ACY.

The invention discloses a method of producing L-alanyl-L-glutamine from recombinant escherichia coli, wherein the method of producing L-alanyl-L-glutamine from recombinant escherichia coli is follows: in an aqueous solution with a determined pH value, acting on free L-glutamine and L-alanine methyl ester hydrochloride to generate L-alanyl-L-glutamine, recombining a gene segment of the protein with the amino-acid ester acyltransferase of the L-alanyl-L-glutamine into a carrier and transferring into host bacteria, thereby obtaining the host bacteria with the recombinant DNA and capable of strengthening the activity of an L-alanyl-L-glutamine biological synthesis system. The method comprises the following steps: (1) culturing a larger number of recombinant escherichia coli cells for expressing the amino-acid ester acyltransferase; (2) excessively expressing the amino-acid ester acyltransferase in the step (1); and (3) taking the amino-acid ester acyltransferase in the step (2) as a crude enzyme source, adding the crude enzyme source into a buffer solution containing L-glutamine and L-alanine methyl ester hydrochloride substrate amino acid to react, thereby realizing efficient production of the L-alanyl-L-glutamine.

The present invention relates to methods of producing industrial products from plant lipids, particularly from vegetative parts of plants. In particular, the present invention provides oil products such as biofuel, and processes for producing these products, as well as plants having an increased level medium chain fatty acids such as lauric acid and myristic acid. In one particular embodiment, the present invention relates to combinations of modifications in a fatty acid thioesterase and one or more acyltransferases. In an embodiment, the present invention relates to a process for extracting lipids. In another embodiment, the lipid is converted to one or more hydrocarbon products in harvested plant vegetative parts to produce alkyl esters of the fatty acids which are suitable for use as a renewable biofuel.