63 results about "Coenzyme A biosynthesis" patented technology

The invention provides a genetically engineered bacterium for producing O-succinyl-L-homoserine as well as a construction method and application of the genetically engineered bacterium. In a chassis bacterium genome, the expression of a 2-ketoglutaric acid decarboxylase encoding gene sucA is enhanced, and the expression of a succinyl-coenzyme A synthetase encoding gene sucD is weakened, so that the supply of succinyl-coenzyme A is increased; the method comprises the following steps: increasing the NADPH (Nicotinamide Adenine Dinucleotide Phosphate) reducing capacity and ATP (Adenosine Triphosphate) energy supply of a chassis bacterium, increasing DNA (Deoxyribose Nucleic Acid) in combination with a transcription dual regulatory factor ompR to improve the stress resistance of escherichia coli under high osmotic pressure, and introducing an overexpression plasmid containing a homoserine transsuccinylase coding gene metA to construct the genetically engineered bacterium for producing O-succinyl-L-homoserine. The engineering strain obtained through a systematic metabolic engineering modification strategy can realize effective accumulation of OSH, the shake flask yield of OSH reaches 19.8 g / L, the fed-batch fermentation yield of a 5L fermentation tank reaches 110.5 g / L, the sugar-acid conversion rate reaches 52.6%, and a foundation is laid for subsequent construction of high-yield OSH engineering bacteria.

The application belongs to the technical field of genetic engineering, and discloses a kind of saccharomyces cerevisiae for producing rare ginsenoside by seaweed biomass and a construction method and application thereof.The saccharomyces cerevisiae has the following characteristics: overexpression of agarase, neojuncanohydrolase, hydroxymethylglutaryl coenzyme A reductase, isopentenyl diphosphate delta isomerase, dammaradienol synthase, protopanaxadiol synthase, cytochrome P450 reductase, protopanaxatriol synthase and glycosyltransferase.The application combines enzymolysis of seaweed biomass with fermentation of rare ginsenoside, which not only endows wild-type yeast with the ability to degrade seaweed biomass that it originally does not have, but also effectively improves the yield of squalene and downstream terpenes (rare ginsenoside Rh1) by overexpression of tHMG1 and IDI1.The saccharomyces cerevisiae can effectively utilize seaweed biomass to obtain squalene and rare ginsenoside Rh1, and has the characteristics of simplicity, economy and ecological friendliness, can convert cheap biomass into high-value products, and provides a way for the development of marine bioeconomy.

The invention discloses a Chimonobambusa chensinensis CuCCoAOMT1 gene as well as a protein and application thereof, belongs to the technical field of plant genetic engineering, and particularly relates to a Chimonobambusa chensinensis caffeoyl-coenzyme A-O-methyltransferase CuCCoAOMT1 gene, and a nucleotide sequence of the CuCCoAOMT1 gene is shown as SEQ ID NO.1. The invention further discloses a preparation method of the CuCCoAOMT1 gene. The invention further discloses application of the CuCCoAOMT1 gene in regulation and control of the drought resistance of gramineous plants. By overexpressing the caffeoyl-coenzyme A-O-methyltransferase CuCCoAOMT1 gene in the rice, the lignin content in the rice can be effectively increased, the thickening of rice cell walls is promoted, the drought resistance of rice plants is improved, and a new gene resource is provided for plant material property improvement gene engineering.

The invention discloses yarrowia lipolytica with high yield of beta-carotene as well as a construction method and application of the yarrowia lipolytica. According to the yarrowia lipolytica engineering bacterium, geranyl diphosphate synthase gene xdGPS, phytoene dehydrogenase gene CarB, phytoene cyclization / phytoene synthesis bifunctional enzyme mutant gene GarRPY27R, acetyl-coenzyme A synthetase mutant gene ACSL641P and pyruvate ferredoxin oxidoreductase gene nifJ are integrated and expressed on a chromosome, and the yarrowia lipolytica engineering bacterium is obtained. Meanwhile, a 3-hydroxy-3-methylglutaryl CoA reductase gene HMGR (3-hydroxy-3-methylglutaryl CoA reductase) derived from the yarrowia lipolytica strain is subjected to overexpression; the yield of the beta-carotene is increased to 13.186 g / L, and the production efficiency is 0.14 g / L / h.

The invention discloses recombinant saccharomyces cerevisiae for producing protopanoxadiol by metabolizing glycerol and a construction method thereof, and the construction method comprises the following steps: introducing a dammarendiol synthase DS gene expression cassette, a PPD synthase and reductase fusion protein PPDS-ATR1 gene expression cassette and a squalene oxidase ERG1 gene expression cassette into saccharomyces cerevisiae to obtain a recombinant bacterium 1; introducing a glycerol transport protein CjFPS1 gene expression cassette, a glycerol dehydrogenase OpGDH gene expression cassette, a dihydroxy acetone kinase DAK1 gene expression cassette and a 3-hydroxy-3-methylglutaryl coenzyme A reductase HMGR gene expression cassette into the recombinant bacterium 1 to obtain a recombinant bacterium 2; introducing citric acid lyase AnACLa and AnACLb gene expression cassettes and citric acid transporter protein YHM2 and CTP1 gene expression cassettes into the recombinant bacterium 2 to obtain a recombinant bacterium 3; experiments prove that the yield of the recombinant bacteria 2, 3PPD is 124 mg / L and 220 mg / L.

Described is a recombinant organism or microorganism which is capable of enzymatically converting acetyl-CoA into isobutene, (A) wherein in said organism or microorganism: (i) acetyl-CoA is enzymatically converted into acetoacetyl-CoA, (ii) acetoacetyl-CoA is enzymatically converted into 3-hydroxy-3-methylglutaryl-CoA, (iii) 3-hydroxy-3-methylglutaryl-CoA is enzymatically converted into 3-methylglutaconyl-CoA, (iv) 3-methylglutaconyl-CoA is enzymatically converted into 3-methylcrotonyl-CoA, and (v) wherein said 3-methylcrotonyl-CoA is converted into isobutene by: (a) enzymatically converting 3-methylcrotonyl-CoA into 3-methylcrotonic acid which is then further enzymatically converted into said isobutene; or (b) enzymatically converting 3-methylcrotonyl-CoA into 3-hydroxy-3-methylbutyryl-CoA which is then further enzymatically converted into 3-hydroxy-3-methylbutyric acid which is then further enzymatically converted into 3-phosphonoxy-3-methylbutyric acid which is then further enzymatically converted into said isobutene; (B) wherein said recombinant organism or microorganism has an increased pool of coenzyme A (CoA) over the organism or microorganism from which it is derived due to: (i) an increased uptake of pantothenate; and / or (ii) an increased conversion of pantothenate into CoA. Moreover, described is the use of such a recombinant organism or microorganism for the production of isobutene. Further, described is a method for the production of isobutene by culturing such a recombinant organism or microorganism in a suitable culture medium under suitable conditions.

The invention discloses application of thiamine in preparation of a product for treating subclinical ketosis of dairy cows, and belongs to the field of biological medicines. Tests prove that compared with dairy cows suffering from subclinical ketosis (SCK), by adding thiamine, the content of beta-hydroxybutyric acid (BHBA), the number of bacteria in milk and the number of somatic cells can be remarkably reduced, and the milk yield, the milk fat content, the acetic acid content and the A / P ratio are remarkably increased; the microbial diversity of rumen and excrement of the dairy cow can be adjusted; a metabonomics result shows that metabolite up-regulated after the thiamine is added mainly focuses on pyruvic acid metabolism, glycolysis or gluconeogenesis, tyrosine metabolism, glycerophospholipid metabolism and biosynthetic pathways of phenylalanine, tyrosine and tryptophan. The conclusion is that when thiamine is added into the SCK dairy cow, synthesis and catabolism of BHBA of an organism can be effectively regulated and controlled by adjusting the content of succinyl-coenzyme A, then subclinical ketosis is relieved, rumen fermentation is promoted, and therefore the milk yield and the milk quality are improved.

The invention discloses an application of a succinyl-coenzyme a synthetase subunit alpha (SUCLG1) as a drug target in prevention and treatment of renal fibrosis, belongs to the technical field of biological medicines, and proves that the expression of SUCLG1 in renal fibrosis tissues is down-regulated, renal fibrosis occurs after the SUCLG1 in normal kidney tissues is selectively knocked down, and the renal fibrosis cannot be affected. And the degree of renal fibrosis can be relieved by over-expression up-regulation of SUCLG1 in renal tubular epithelial cells, so that the SUCLG1 can be determined to be used as an important target for preventing and treating renal fibrosis, and drugs for regulating and increasing the expression of SUCLG1 can be researched and developed in the field, so that the drugs are used for intervening the occurrence and development of renal fibrosis, and new potential therapeutic targets and therapeutic strategies are provided for renal fibrosis.

The present disclosure relates to the use of a switch for the production of heterologous non-catabolic compounds in microbial host cells. In one aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured under aerobic conditions followed by microaerobic conditions, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions. In another aspect, provided herein are genetically modified microorganisms that produce non-catabolic compounds more stably when serially cultured in the presence of maltose followed by the reduction or absence of maltose, and methods of producing non-catabolic compounds by culturing the genetically modified microbes under such culture conditions.

The present disclosure describes a recombinant yeast host cell having a first engineered metabolic pathway to convert fructose-6-phosphate (F-6-P) into acetyl-coenzyme A. The first engineered metabolic pathway comprises: a genetic modification for expressing a heterologous phosphoketolase capable of converting fructose-6-phosphate into erythrose-4-phosphate, and at least one genetic modification for decreasing native 6- phosphofructo-2-kinase activity classified under Enzyme Commission No. 2.7.1.105. Optionally, the recombinant yeast host cell comprises a second engineered metabolic pathway for convert acetyl-coA into a fermentation product. The present disclosure further describes a process for making a fermentation product. The process comprises contacting the recombinant yeast host cell described herein with a carbohydrate source under a condition allowing the conversion of at least a part of the carbohydrate into a fermentation product.

The invention relates to ribozyme for labeling biotin on target RNA (Ribonucleic Acid) as well as a screening method and application thereof. The RNA labeling technology is one of core tools for modern molecular biology research and clinical diagnosis. A specific marker is covalently linked to a target RNA molecule, so that accurate tracking, efficient detection and functional analysis of RNA can be realized. A biotin-streptavidin system becomes one of the technical routes which are most widely applied due to extremely high affinity and signal amplification capability of the biotin-streptavidin system. At present, common biotin-labeled RNA technical methods mainly comprise a chemical solid-phase synthesis method, an enzymatic method and a chemical labeling method. However, the methods have the limitations of poor site specificity, low efficiency, complex manipulation and the like in the use process. The engineering ribozyme for catalyzing RNA biotinylation is obtained by taking 12: 0 biotin coenzyme A as a biotin donor through an in-vitro screening technology. The ribozyme provided by the invention can efficiently and specifically carry out biotin labeling on target RNA, the reaction process is simple, and the labeling activity is up to 90% or above.

This invention belongs to the fields of fermentation and genetic engineering technology, and provides a high-yield hexanoic acid-producing engineered *Escherichia coli* strain, its construction method, and its applications. This engineered strain is *Escherichia coli*... E. coli w3110 was the starting strain, overexpressing the acetyl-CoA transferase gene. act and β-ketothiolase gene bktB Overexpression of trans-enoyl-CoA reductase gene ter 3-Hydroxybutyryl-CoA dehydrogenase gene hbd, and 3-hydroxybutyryl coenzyme A dehydratase gene crt Or, simultaneously not expressing the pyruvate formate lyase gene. pflB or lactate dehydrogenase gene ldhA This engineered bacteria can convert glucose into hexanoic acid, a high-value product, with a yield of 4.968 g / L after 18 hours of fermentation. It adopts aerobic fermentation, resulting in rapid cell growth, a short fermentation cycle, and a high acid production rate. The fermentation process is simple, easy to control, and has low production costs, which is conducive to its promotion and application in industrial production.

The invention provides a genetic engineering strain of high-yield immunosuppressant mycophenolic acid as well as a preparation method and application of the genetic engineering strain. The invention firstly provides a method for constructing the penicillium with the effect of high yield of mycophenolic acid. The method comprises the step of replacing an indigenous promoter of acyl-coenzyme A oxidase PbACOX323 protein participating in beta oxidation in peroxisome of a penicillium strain with a promoter PgpdA of 3-phosphoglycerol dehydrogenase with an amino acid sequence of SEQ ID NO: 1. According to the method disclosed by the invention, the genetically engineered bacterium with high yield of mycophenolic acid is constructed, the yield of mycophenolic acid is positively increased by replacing a promoter of acyl-coenzyme A oxidase PbACOX323, and the method has a good industrial application value.

The invention discloses a compound enzyme preparation-containing nutritional feed for regulating and controlling intramuscular fat of black pigs and a preparation method of the compound enzyme preparation-containing nutritional feed, and relates to the technical field of livestock and poultry breeding feeds. The invention discloses a black pig skeletal muscle cell fat directional transport feed which is prepared from the following components in parts by weight: 850-920 parts of basal feed, 5-12 parts of fat directional deposition compound enzyme group, 25-40 parts of amino acid-enzymatic synthesis substrate combination and 2-6 parts of lipolytic enzyme specific inhibitor. According to the black pig skeletal muscle cell fat directional transport feed, fat precursor directional transport is realized through the compounded fat directional deposition compound enzyme group, a fat synthesis pathway targeting black pig skeletal muscle cells and lipoprotein lipase; acetyl-coenzyme A carboxylase and fatty acid synthetase catalyze fat synthesis step by step, only intramuscular fat generation pathways are activated, subcutaneous and abdominal fat deposition paths are blocked, the technical defects that fat deposition of an existing single enzyme preparation is non-directional, and the body fat rate is caused to be abnormally increased are overcome, meanwhile, an exclusive amino acid-enzymatic synthesis substrate combination is matched, and the body fat synthesis efficiency is improved. Sufficient adaptive substrates are supplied to enzyme group catalytic reaction, and the problem that a single enzyme preparation is insufficient in catalytic efficiency is solved.

The invention provides a method for producing DL-2, 4-dihydroxy-3, 3-dimethyl butyric acid (DL-pantoic acid) and / or DL-3-hydroxy-4, 4-dimethyl oxacyclopentane-2-one (DL-pantolactone), in particular to a method for producing DL-2, 4-dihydroxy-3, 3-dimethyl butyric acid and / or DL-3-hydroxy-4, 4-dimethyl oxacyclopentane-2-one (DL-pantolactone) by condensation of hydroxypivalaldehyde and formyl-CoA of C1 source, in particular to a method for producing DL-2, 4-dihydroxy-3, 3-dimethyl oxacyclopentane-2-one (DL-pantoic acid) and / or DL-3-hydroxy-4, 4-dimethyl oxacyclopentane-2-one (DL-pantoic acid). The invention relates to a method for preparing 2, 4-dimethyl oxacyclopentane-2-one. The present invention also provides a genetically engineered microorganism for use in the production of DL-2, 4-dihydroxy-3, 3-dimethylbutyric acid and / or DL-3-hydroxy-4, 4-dimethyl oxacyclopentane-2-one, and also provides a genetically engineered microorganism for use in the production of DL-3-hydroxy-4, 4-dimethyl oxacyclopentane-2-one. In addition, the key enzyme has been engineered to improve its performance in the production of DL-pantoic acid or DL-pantolactone.

The invention relates to a preparation method and application of 6-methyl-3, 5-dioxocapryloyl coenzyme A. According to the preparation method, 2-methylbutyric acid serves as a starting raw material, the 6-methyl-3, 5-dioxocapryloyl coenzyme A is chemically synthesized through the steps of activation, condensation, coenzyme A coupling and the like, and the structure and purity of the 6-methyl-3, 5-dioxocapryloyl coenzyme A are identified through the technologies of LC-MS / MS, 1HNMR and the like. The nine-carbon coenzyme A is successfully prepared and identified through a chemical synthesis method for the first time, the core problem that an intermediate is unstable and cannot be obtained in vitro is solved, and a stable substrate is provided for in-vitro acyl transfer; the QS-21 in-vitro acyl transfer reaction is realized for the first time, an in-vitro catalytic system of a chemical synthesis intermediate and an insect cell expression enzyme is established, and a foundation is laid for development of a QS-21 total synthesis process.

The invention provides recombinant halomonas, a construction method of the recombinant halomonas and application of the recombinant halomonas in production of polyhydroxyalkanoate. The recombinant halomonas is used for expressing the 2-keto acid decarboxylase or the 2-keto acid decarboxylase mutant. According to the invention, a 2-ketonic acid decarboxylase mediated 3HV synthesis path is constructed, alpha-ketoglutaric acid in tricarboxylic acid circulation can be directly converted into 3-hydroxyvaleryl coenzyme A, de novo synthesis of a 3HV monomer is realized, a conventional carbon source can be efficiently utilized to synthesize 3HV, and the traditional technical bottleneck of dependence on an exogenous propionic acid or valeric acid precursor is broken through; the 3HV with a higher proportion can be obtained by regulating and controlling a metabolic pathway under the condition of not adding an exogenous precursor, so that the fermentation process is remarkably simplified, and the raw material cost and the production complexity are reduced.

The invention relates to a 5-carboxyl-2-pentenoyl-coenzyme A reductase mutant and application thereof, and belongs to the technical field of biological engineering. Aiming at the problem of low cofactor binding efficiency of 5-carboxyl-2-pentenoyl-coenzyme A reductase in a reverse adipic acid degradation pathway, rational prediction and site-directed mutagenesis experiments are carried out by taking a gene for expressing the 5-carboxyl-2-pentenoyl-coenzyme A reductase as a research object, structural optimization is carried out on a binding site of the enzyme, and the cofactor binding efficiency of the 5-carboxyl-2-pentenoyl-coenzyme A reductase in the reverse adipic acid degradation pathway is improved. According to the method disclosed by the invention, the adipic acid is more suitable for the characteristics of cofactors, so that the binding capacity of the enzyme to the cofactors is enhanced, the activity of catalyzing the reaction of 5-carboxyl-2-pentenoyl-coenzyme A to generate adipoyl coenzyme A is improved, and the yield of the adipic acid is further improved.

The invention discloses a construction method of a saccharomyces cerevisiae mutant with high yield of 3-hydracrylic acid, and belongs to the technical field of biology. SKN7 gene, PGD1 gene, STB3 gene, YAP1 gene, RIM101 gene, CIN5 gene, FHL1 gene, HAA1 gene, CRZ1 gene, NRG1 gene or PDR1 gene of saccharomyces cerevisiae are mutated and screened to obtain a saccharomyces cerevisiae mutant strain with high yield of malonyl-coenzyme A, and then 3-hydracrylic acid production plasmid is introduced into the obtained saccharomyces cerevisiae mutant strain to obtain the saccharomyces cerevisiae mutant with high yield of 3-hydracrylic acid. Compared with control bacteria, the yield of 3-hydracrylic acid of the saccharomyces cerevisiae mutant is remarkably increased, a technical basis is provided for directional transformation of saccharomyces cerevisiae engineering strains, applicable strains can be provided for carbon fiber production, and industrial production of 3-hydracrylic acid is promoted.

The invention provides an application of up-regulated human enoyl coenzyme A delta isomerase 2 (ECI2) in preparation of a medicine for protecting a cardiac function. It is found for the first time that up-regulation of ECI2 promotes degradation of crotonyl coenzyme A by consuming a crotonyl donor (crotonyl coenzyme A) and effectively inhibits crotonyl modification, so that the heart function and myocardial hypertrophy can be effectively improved, and then the heart function protection effect on pressure overload heart diseases and heart failure is achieved. The above findings indicate a huge potential of upregulating ECI2 in the prevention and / or treatment of pressure overload heart diseases and heart failure.

The application discloses a kind of recombination of Pseudomonas putida for producing L-lactic acid using whole-component lignocellulose hydrolysate and its construction method and application, belong to the field of metabolic engineering.The application uses Pseudomonas putida KT2440 as chassis, realizes carbon flow to pyruvic acid convergence and prevents product degradation by knocking out aceEF 、 ppsA And lldD Gene;By in-situ replacement, introduce optimized Lactobacillus plantarum ldhL1 Reconfiguration L-lactic acid synthesis pathway;By integration xylD-yjhH Tandem expression cassette enables xylose utilization;And by quorum sensing promoter P Rox306 Overexpression maeB Realize funnel conversion of lignin monomer.The application innovatively uses lignin derivatives in hydrolysate as natural donor of endogenous acetyl-coenzyme A, makes up lethal growth defect caused by knocking out aceEF The engineering bacteria of the application realize the synchronous high-efficiency conversion of glucose, xylose and aromatic monomer under the condition of not adding exogenous auxiliary carbon source and chemical inducer, which greatly reduces the industrial fermentation cost.

The application belongs to the technical field of diagnostic markers, and particularly relates to a pathological diagnosis biomarker combination for adrenal Cushing syndrome and application. The pathological diagnosis biomarker combination comprises low density lipoprotein receptor (LDLR), 3-hydroxy-3-methylglutaryl coenzyme A synthetase 1 (HMGCS1) and cytochrome P450 11B1 (CYP11B1). The area under the ROC curve (AUC) of the pathological diagnosis biomarker combination can reach 0.881 (95% confidence interval (CI): 0.775-987; sensitivity: 85.7%; specificity: 85.0%), which indicates that the combination of LDLR, HMGCS1 and CYP11B1 as the pathological diagnosis biomarker combination for adrenal Cushing syndrome has high accuracy.

The application discloses a blood-activating herb naringenin synthase gene GlCHS, a product coded by the gene and application thereof, and belongs to the field of genetic engineering. The blood-activating herb naringenin synthase gene GlCHS is cloned from a blood-activating herb, the nucleotide sequence of the gene is shown as SEQ ID No. 1, and the amino acid sequence of the product coded by the gene is shown as SEQ ID No. 2. The blood-activating herb naringenin synthase is expressed heterogeneously in a host cell, and the enzyme has the function of catalyzing naringenin to be generated by taking p-coumaroyl coenzyme A and malonyl coenzyme A as substrates. By using the application, the content of naringenin in the blood-activating herb can be improved and naringenin can be synthesized in vitro in a medicinal plant by using genetic engineering technology.

The invention belongs to the technical field of biological medicine and tumor immunotherapy, and discloses application of 3-oxo-valproic acid coenzyme A in preparation of a medicine for inhibiting lung adenocarcinoma. As a specific agonist of GIMAP1, the 3-oxo-valproic acid coenzyme A is used for activating the GIMAP1, enhancing inhibition of lung cancer cell progress and enhancing activation of a tumor immune microenvironment, so that the lung adenocarcinoma is inhibited, and the lung adenocarcinoma is inhibited. The invention has the following beneficial effects: 1, strong targeting property: 3-oxo-valproate coenzyme A is specifically combined with GIMAP1, so that a mediated immunomodulatory pathway is accurately activated, and wide influence on normal cells is avoided; 2, immune microenvironment remodeling: the anti-tumor immune cell infiltration is remarkably improved, the lung adenocarcinoma immunosuppression microenvironment is reversed, and the immunotherapy sensitivity is improved; 3, tumor progression is inhibited, lung cancer cell proliferation is effectively inhibited, and the tumor invasion ability is reduced; 4, the source is natural: as an intestinal microbial metabolite derivative, the safety is high, and the side effect is lower than that of a chemically synthesized medicine.

The application relates to the field of biological medicines, and discloses application of coenzyme A in preparation of a myocardial cell ferroptosis inhibitor. The application first finds that coenzyme A has the function of inhibiting myocardial cell ferroptosis, and the function is realized by combining with IGFBP2 protein, so that a new drug for targeted inhibition of myocardial cell ferroptosis is expected to be developed, and a new theoretical basis and potential intervention strategy are provided for myocardial injury repair.

The invention discloses application of a Reg4 gene in treatment of metabolism-related fatty liver, and relates to the technical field of biomedicine. The Reg4 gene is applied to the aspect of treating obesity and metabolism-related fatty liver; according to the application, by increasing expression of the Reg4 gene, liver lipid deposition, liver cell ballooning and inflammatory infiltration related to the metabolism-related fatty liver are inhibited; the application is realized by inhibiting the processes of de novo synthesis of fat in the liver, liver lipid uptake and fatty acid beta oxidation. The step of inhibiting the de novo synthesis of fat in the liver comprises the step of inhibiting the expression of genes of acetyl coenzyme A carboxylase 1, fatty acid synthase and / or stearoyl coenzyme A desaturase 1. The invention definitely proves that the Reg4 gene has an obvious effect of improving obesity and metabolism-related fatty liver, and can relieve hepatic fatty degeneration and inflammation.

The invention discloses a high-stability anti-interference free fatty acid detection kit as well as a preparation method and application thereof, the kit comprises a reagent R1 and a reagent R2, the reagent R1 comprises a phosphate buffer solution, acetyl coenzyme A synthetase, coenzyme A, adenosine triphosphate, 4-aminoantipyrine and a cyclodextrin-metal organic framework nano stabilizer with the concentration of 5-20 mg / mL; and the reagent R2 is prepared from a Tris-HCl buffer solution, acetyl coenzyme A oxidase, peroxidase, N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium salt and a cyclodextrin-metal organic framework nano stabilizer with the concentration of 2 to 10mg / mL. Through the CD-MOF nano material with the specific aperture, triple breakthrough of improving the enzyme stability, enhancing the interference resistance of a complex sample and optimizing the detection sensitivity is synchronously realized.

The invention discloses a kit for catalyzing Friedel-Crafts acylation reaction through an enzyme method, application and a preparation method. The kit for catalyzing the Friedel-Crafts acylation reaction by the enzyme method comprises 1) a Friedel-Crafts acylation reaction buffer solution and 2) a thiolase complex MucABC or a functional fragment and a variant thereof. The kit for catalyzing the Friedel-Crafts acylation reaction does not depend on coenzyme A compounds as acyl donors, and can realize regional specific C-acylation modification on various lactam compounds by taking acyl phenyl esters with different chain lengths as the acyl donors under mild conditions; a novel green and efficient biological catalysis tool is provided for Friedel-Crafts acylation reaction, and the method has important value and significance for drug synthesis and chemical production.

An aptamer which selectively produces a detectable signal in the presence of CoA and methods of using the same.

The application provides a recombinant Halomonas, a construction method thereof and application of the recombinant Halomonas in production of polyhydroxyalkanoate. The recombinant Halomonas expresses 2-keto acid decarboxylase or a 2-keto acid decarboxylase mutant. The application constructs a 3HV synthesis path mediated by 2-keto acid decarboxylase, which can directly convert alpha-ketoglutarate in the tricarboxylic acid cycle into 3-hydroxyvaleryl coenzyme A, realizes de novo synthesis of 3HV monomers, can efficiently utilize conventional carbon sources to synthesize 3HV, breaks through the technical bottleneck of traditional dependence on exogenous propionic acid or valeric acid precursors, realizes obtaining of a higher proportion of 3HV under the condition of no need of adding exogenous precursors, and significantly simplifies the fermentation process and reduces raw material cost and production complexity.