207 results about "Acetylcoenzyme A" patented technology

Cytochrome b5 (Cb5) is a haem-binding protein located in the endoplasmic reticulum (ER) and the outer mitochondrial membranes of higher eukaryotes. In higher plants, animals, and fungi, the ER resident Cb5 has been shown to play a role in desaturation of acyl CoA fatty acids. Higher plants Cb5 isoforms from plants such as soybean or Arabidopsis are capable of modulating omega-3 desaturation. Co-expression of certain Cb5 isoforms with FAD3 in a host plant results in increased production of seed oil content as well as altered ratio between different fatty acids. It is also disclosed here that overexpression of Yarrowia ACL enzymes in the plastids of a host plant helps boost the synthesis of acetyl CoA, which in turn, may lead to increased synthesis of fatty acids and enhanced oil accumulation in the seeds.

The invention provides a non-naturally occurring microbial organism having an acetyl-CoA pathway and the capability of utilizing syngas or syngas and methanol. In one embodiment, the invention provides a non-naturally occurring microorganism, comprising one or more exogenous proteins conferring to the microorganism a pathway to convert CO, CO₂ and/or H₂ to acetyl-coenzyme A (acetyl-CoA), methyl tetrahydrofolate (methyl-THF) or other desired products, wherein the microorganism lacks the ability to convert CO or CO₂ and H₂ to acetyl-CoA or methyl-THF in the absence of the one or more exogenous proteins. For example, the microbial organism can contain at least one exogenous nucleic acid encoding an enzyme or protein in an acetyl-CoA pathway. The microbial organism is capable of utilizing synthesis gases comprising CO, CO₂ and/or H₂, alone or in combination with methanol, to produce acetyl-CoA. The invention additionally provides a method for producing acetyl-CoA, for example, by culturing an acetyl-CoA producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding an acetyl-CoA pathway enzyme or protein in a sufficient amount to produce acetyl-CoA, under conditions and for a sufficient period of time to produce acetyl-CoA.

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 an expression system and constructing PHB transgene algae method of rhine chlamydomonas exogenesis gene, which comprises the following parts: promotor, exogenesis-goal gene, screening badge and rhine chlamydomonas acceptor algae strain, wherein the method comprises the following: A, selecting and cultivating acceptor algae strain; B, cloning key enzyme gene by PHB biosynthesizing; C, expressing carrier of PHB biosynthesizing key enzyme rhine chlamydomonas; D, using PHB biosynthesizing key enzyme to lead in rhine chlamydomonas from fungus Bacillus alcaligenes with bead-milling method, gene-gun method or electrization; obtaining bivalence or transgene Chlamydomonas of producing PHB by screen selecting and molecule detecting. The poly-hydroxybutyric acid is compounded by acting in conjunction of PHB compounding key enzyme of phbA, phbB and phbC gene coding, which uses acetylcoenzyme A as substrate by photosynthesis. The method simplifies the operating, which reduces cost.

The invention provides a biological method for synthesizing alpha-pinene or beta-pinene from acetyl coenzyme A and a reconstitution cell capable of synthesizing the alpha-pinene or beta-pinene, wherein the acetyl coenzyme A is obtained from a simple starting material like glucose. The method for synthesizing the alpha-pinene or beta-pinene by adopting a biological process comprises the following steps: transferring acetyl coenzyme A acyltransferase, 3-hydroxy-3-methyl glutaryl coenzyme A synthase, hydroxymethyl glutaryl coenzyme A reductase, mevalonic acid kinase, mevalonic acid-5-phosphokinase, mevalonic acid-5-diphosphonic acid decarboxylase, isopentenylpyrophosphate isomerase, geranyldiphosphate synthetase and pinene synthase into an appropriate host cell by adopting metabolic engineering technology, and culturing the obtained reconstitution cell, so that the target product, namely alpha-pinene or beta-pinene can be obtained.

The invention provides a method for constructing a gene engineering strain to enhance the resistance of a 1, 3-propanediol (PDO) production strain, which belongs to the technical field of biochemical engineering and comprises the step of: introducing Beta-ketothiolase PhbA, NADPH-dependent acetoacetyl CoA, PhbB and polyhydroxyalkanoates synthases, PhbC gene to a wild bacteria which can produce PDO to accumulate high concentration of PHB while the bacterial strain produces PDO so as obviously improve the tolerance of the bacterial strain to glycerol and 3-hydroxypropionaldehyde; the method can also be used to co-produce PHB and PDO. The method has the advantages that the constructed gene engineering strain improves the resistance of the bacterial strain to the high concentration of glycedrol and the intermediate product 3-hydroxypropionaldehyde and reduces the abnormal fermentation in industrial production; in addition, high concentration of PHB can be accumulated while PDO is produced, and PHB can be used as a separated product, thereby promoting the added valve of products and the utilization rate of materials and reducing the production cost.

The invention provides a recombinant escherichia coli strain, a preparation method of the recombinant escherichia coli strain and a method for biologically synthesizing poly-3-hydroxypropionic acid from acetyl coenzyme A. The method comprises the following steps: with glucose or glycerin and the like as carbon sources, commonly over-expressing endogenous or exogenous acetyl coenzyme A carboxylase genes (acc) and propionyl coenzyme A synthetase genes (prpE) as well as exogenous malony coenzyme A reductase genes (mcr) and polyhydroxyalkanoate synthetase genes (phaC) in proper host cells (such as escherichia coli), and degrading intermediate products by virtue of glucose so as to biologically synthesize the poly-3-hydroxypropionic acid, wherein acetyl coenzyme A is finally obtained from simple starting materials such as the glucose and the like.

The invention discloses an acetyl coenzyme A acetyltransferase gene RKAcaT2 and application thereof. The nucleotide sequence of the gene is shown in SEQ ID NO:1, the amino acid sequence encoded by thegene is shown in SEQ ID NO:2, the gene is a key enzyme gene synthesized by carotenoid in Rhodosporidium kratochvilovae YM25235 and has the functions of the acetyl coenzyme A acetyltransferase, and the carotenoid produced by the Rhodosporidium kratochvilovae YM25235 can be controlled; microorganisms are transformed through the genetic engineering means so as to increase the yield of carotenoid inthe microorganisms, and a foundation is laid for large-scale commercial production of the carotenoid.

The invention discloses a method for positioning and synthesizing terpenoids by means of the Yarrowia lipolytica way. The method comprises the steps that an acetoacetyl coenzyme A thiolase, HMG-CoA synthetase and HMG-CoA reductase in the synthetic route of mevalonic acid in Yarrowia lipolytica are over-expressed and positioned to the peroxisome, and engineering bacteria MP1 are obtained; on the basis of the engineering bacteria MP1, the alpha-farnesene synthesis way is expressed, and engineering bacteria FP1 are obtained; or, the beta-carotene synthesis way is expressed, and engineering bacteria CP1 are obtained; or, the linalool synthesis way is expressed, engineering bacteria LP1 are obtained; the bacteria FP1 or CP1 or LP1 can synthesize the terpenoids under the aerobatic condition by means of a culture medium containing fatty acid or grease, and the yield of the synthesized terpenoids is higher compared with a traditional method. According to the method, fatty acid, grease and cheap food waste oil can be used for generating mevalonic acid downstream terpenoids, and the considerable application prospect and the economic value are achieved.

The invention discloses engineering bacteria for producing trans-4-hydroxy-L-proline and a construction method and an application thereof. The construction method of the engineering bacteria provided by the invention includes the steps of A1) and A2): A1) introducing a L-proline-4-hydroxylase gene, a glutamate-5-kinase gene and a glutamate-5-semialdehyde dehydrogenase gene into a receptor cell; and A2) knocking an alpha-ketoglutaricdehydrogenase gene, an isocitratlyase gene or a proline dehydrogenase gene of the receptor cell out, or replacing a pyruvic oxidase gene of the receptor cell with an acetylcoenzyme A synthetase gene; and carrying out a reaction of the recombinant cell and a substrate to obtain the trans-4-hydroxy-L-proline. Experiments prove that the production method of the trans-4-hydroxy-L-proline can be used for production of the trans-4-hydroxy-L-proline.

The invention belongs to the technical field of microbial fermentation and relates to a method for efficiently preparing natural abscisic acid through adding acetylcoenzyme A as precursor substances for strains to synthesize the abscisic acid in a flowing way in the fermentation process and simultaneously adding biosurfactant tween-20 for improving the dissolved oxygen condition of the fermentation liquid. The method comprises the following steps that fungi capable of producing the natural abscisic acid are cultured in a first stage liquid culture medium; the cultured first stage seed liquid is inoculated onto a second stage liquid culture medium for culture, supplementing materials are added to a third stage liquid culture medium in the flowing way, simultaneously, the tween-20 is added,and the acetylcoenzyme A is added in the flowing way for fermentation culture; and the obtained abscisic acid is collected from the fermentation culture liquid. The method has the advantages that theoxygen utilization rate of the strains and the acid yield can be improved, so the strains can be used for producing the abscisic acid at higher substrate conversion rate and higher product synthesis velocity, the yield and the production efficiency are improved, the energy consumption is reduced, and the production cost is reduced.

The invention relates to a free fatty acid determination reagent kit which comprises dual reagents. The reagent 1 comprises Good's buffer solution (pH(potential of hydrogen)=7.0), coenzyme A, ATP (adenosine triphosphate), acetylcoenzyme A synthetase, magnesium chloride (MgCL2), Trinder bound constituent, tween 80 and liquid biological preservative/Proclin 300, and the reagent 2 comprises Good's buffer solution (pH=7.0), acetylcoenzyme A oxidase (ACOD), peroxidase (POD), Trinder bound constituent, tween 80 and liquid biological preservative/Proclin 300. The free fatty acid determination reagent kit has the remarkable advantages that the reagent kit is directly used for an automatic biochemical analyzer and is simple; and the adopted automatically separated and purified acetylcoenzyme A synthetase and acetylcoenzyme A oxidase (ACOD) have high specificity on free fatty acid in serum.

The invention discloses a construction method and an application of an electrochemical Faraday cage immunosensor for detecting the activity of histone acetyltransferase. The construction method comprises the following steps: (1) preparing peptide/Au: respectively taking acetyltransferase p300, polypeptide and acetyl-CoA, fully mixing the taken substances in PBS (0.1 M, pH 7.0), incubating the obtained solution in a constant-temperature water bath, taking and dropwise applying a catalytic reaction to the surface of a gold electrode, and incubating the gold electrode in a 4 DEG C refrigerator; (2) preparing MB&AuNPs@GO-Ab: mixing HAuCl4 and CTAB with water, adding ascorbic acid to the obtained reaction mixture, adding NaOH to obtain CTAB covered AuNPs, centrifuging and purifying the CTAB covered AuNPs, dispersing the purified CTAB covered AuNPs in an equal amount of water, adding GO to the obtained solution, performing ultrasonic dispersion, standing the dispersed solution for later use,adding an acetyl antibody, performing incubation, adding MB, and performing vibration and uniform mixing; and (3) producing the electrochemical Faraday cage immunosensor: taking and dropwise applyingthe MB&AuNPs@GO-Ab to the surface of the peptide/Au, performing incubation at room temperature, and placing the produced sensor in the PBS (0.1 M, pH 7.0) to carry out electrochemical SWV test.

The invention discloses a preparation method of an electrochemical biosensor for simultaneously detecting HAT and TdT and application thereof. The method comprises: step one, adding MCH dropwise ontoa hairpin DNA modified electrode surface induced by Hg<2+> and marking the processed electrode as Electrode 1; step two, taking acetyltransferase p300, polypeptide and acetylcoenzyme A respectively and mixing the acetyltransferase p300, the polypeptide and the acetylcoenzyme A fully in a phosphoric acid buffer solution (PBS, 10 mM, pH 7.0), dropping and coating the mixed solution on the surface ofthe Electrode1, and marking the obtained substance as Electrode2; step three, dropping and coating an Exo I solution on the surface of the Electrode2, carrying out incubation at a room temperature, dropping a TdT reaction solution on the electrode surface, and marking the obtained substance as Electrode3; and step four, adding Cu<2+> on the electrode surface obtained in the step three, dropping an ascorbic acid solution on the electrode surface, marking the obtained substance as Electrode 4, and detecting an electrochemical response in PBS electrolyte solution with parameters of 0.1M and pH7.0. The preparation method has the following advantages: the specificity and sensitivity are high; the detection speed is fast; the activities of two kinds of enzymes can be detected simultaneously; and the result is accurate and reliable and the cost is low.

The invention discloses a genetic engineering bacterium for producing glycollic acid by acetic acid, and a building method and application thereof. A method for preparing the genetic engineering bacterium for producing glycollic acid provided by the invention comprises the following steps of improving the expression and/or activity of acetylcoenzyme A synthetase, phosphotransacetylase, acetokinase, citrate synthase, isocitrate lyase, isocitrate dehydrogenase kinase and glyoxylate reductase in the recipient bacterium; reducing the expression and/or the activity of malate synthase, glycolate oxidase and isocitrate lyase repressor protein in the recipient bacterium, so that the engineering bacterium for producing the glycollic acid is obtained. The recipient bacterium is a bacterium capable of using acetic acid as a carbon source for growth. The prepared genetic engineering bacterium uses acetic acid as the carbon source for producing the glycollic acid; in addition, the production quantity and yield of the glycollic acid in shaking culture can reach the relatively high level; relatively good industrial application prospects are realized.