42 results about "Amidohydrolase" patented technology

The invention discloses a group of amide hydrolase mutants capable of efficiently degrading ochratoxin A as well as a coding gene, a recombinant vector and application of the amide hydrolase mutants, and belongs to the technical field of enzyme engineering. The invention also provides application of the mutant of the group of recombinant amide hydrolase in detoxification of ochratoxin A in beer or corn flour polluted by ochratoxin A. The activity of the mutant on ochratoxin A is obviously improved compared with the activity of the existing recombinant amide hydrolase ADH3; beer or corn flour containing ochratoxin A is treated for 10-15 minutes, the degradation rate of ochratoxin A can reach 100%, and the degradation efficiency is high in the field.

The invention discloses an ochratoxin A degrading enzyme mutant and application thereof. The invention provides mutant protein of amidase MiADH, which is any one of the following changes: the amino acid sequence of the amidase MiADH as shown in SEQ ID NO: 1 is subjected to any one of the following changes: a 66th amino acid residue Y is mutated into K, or a 143rd amino acid residue H is mutated into A, or a 194th amino acid residue V is mutated into A, or a 195th amino acid residue L is mutated into A, or a 230th amino acid residue H is mutated into A, or the amino acid sequence of the amidase MiADH is mutated into A, or the amino acid sequence of the amidase MiADH is mutated into K, or the amino acid sequence of the amidase MiADH is mutated into A; or the 324th amino acid residue V is mutated into A, and the protein has the same function; experiments prove that the amide hydrolase MiADH mutant can efficiently degrade OTA, and OTA detoxification can be achieved under the high-temperature condition.

The invention discloses amide hydrolase PwADH / DM for degrading ochratoxin A as well as a coding gene, a recombinant vector and application of the amide hydrolase PwADH / DM, and belongs to the technical field of enzyme engineering. The amide hydrolase is an amide hydrolase protein of a two-site mutant obtained by further modification on the basis of the existing amide hydrolase capable of efficiently degrading ochratoxin A. The invention provides application of the amide hydrolase in hydrolysis of OTA and detoxification of beer and nuts polluted by OTA. Compared with the existing amide hydrolase PwADH, the amide hydrolase has the advantages that the hydrolysis and detoxification efficiency of the amide hydrolase on OTA is obviously improved, and the amide hydrolase is used for treating beer and almond powder samples polluted by ochratoxin A for 10-15 minutes at room temperature, so that the degradation rate of the ochratoxin A can reach 100%.

The invention relates to the technical field of molecular breeding, and discloses an auxin amide hydrolase RhILL1 gene and application of the auxin amide hydrolase RhILL1 gene in regulation and control of China rose flower color, and the nucleotide sequence of the auxin amide hydrolase RhILL1 gene is shown as SEQ ID NO.1. By silence or overexpression of the RhILL1 gene, synthesis of anthocyanin is effectively regulated and controlled, and accurate control of petal color is realized. Researches find that silencing of the RhILL1 gene leads to color fading of petals and reduction of anthocyanin content; and overexpression of the RhILL1 gene prolongs the color holding time of the petals. In addition, the invention also provides a kit containing the RhILL1 gene regulatory sequence, a recombinant plasmid and a corresponding gene editing and transgenic breeding method. The achievements provide a new molecular tool for Chinese rose breeding and flower color improvement, and have important ornamental value and economic significance.

The invention discloses a biological catalytic synthesis method of chiral 2-amino-1-methylenepropanamide and an enzyme catalyst of the chiral 2-amino-1-methylenepropanamide. According to the present invention, one or more key amino acids in the amide hydrolase derived from Rhodococcus erythropolis AJ270 are subjected to mutation to obtain the mutant, the mutant has characteristics of high or reverse enantioselectivity when the hydrolysis reaction of the racemization 2-substituted-2-amino-1-methylene propanamide substrate is catalyzed, and the chiral compound is obtained with the high ee value; and an enantiomer of a wild type amide hydrolase catalytic product can be obtained. A method for preparing chiral 2-substituted-2-amino-1-methylene propanamide by using the mutant through biological catalysis has the characteristics of simplicity and convenience in operation, high reaction efficiency, mild reaction conditions, high enantioselectivity, easiness in product separation and high product purity, and has a very good application prospect.

The invention discloses a substituted heterocyclic compound containing an α-ketone skeleton and a use thereof, and relates to the field of pharmaceutical chemistry, and specifically relates to a substituted heterocyclic compound containing an α-ketone skeleton or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing these compounds and their medical uses, in particular, an application of the substituted heterocyclic compound as a fatty amide hydrolase (FAAH) inhibitor in the preparation of a drug for preventing or treating diseases related to FAAH, including inflammatory diseases, rheumatoid arthritis, hepatitis, liver fibrosis, autoimmune diseases, pain, depression, pain-depression comorbidity, autism, social phobia, Tourette syndrome, neurodegenerative diseases, anxiety post-traumatic stress (PTSD), cannabinoid use disorder, drug withdrawal, and anti-tumor treatment.

The invention belongs to the field of enzyme catalysis, and particularly relates to an amide hydrolase mutant, a immobilized enzyme, a coding gene, an expression plasmid, a preparation method of the amide hydrolase mutant, the immobilized enzyme, the coding gene and the expression plasmid, and application of the amide hydrolase mutant, the immobilized enzyme, the coding gene and the expression plasmid in catalysis of amide hydrolysis. The gene is characterized in that the gene is an amino acid sequence with at least one mutation of L85Y and E209R in a wild amino acid sequence as shown in SEQ ID NO.1. Researches show that the mutant can significantly enhance the catalytic activity of enzyme, and the reaction conversion rate can be effectively increased when the mutant is used for enzymatic amide hydrolysis reaction; for example, the compound is innovatively used for amide hydrolysis of ASG, and has excellent degradation capacity.

The invention discloses a method for synthesizing 2-amino-6-clobird purine based on amide hydrolase, which specifically comprises the following steps: adding the amide hydrolase into a 2-formamido-6-clobird purine aqueous solution, and reacting for 3-5 hours to obtain the 2-amino-6-clobird purine, and the amide hydrolase is self-prepared recombinant amide hydrolase. The 2-amino-6-chlorobird purine prepared by the method provided by the invention is high in purity, low in guanine impurity content, short in reaction time consumption and low in generated wastewater amount. # imgabs0 #

The invention provides an application of an HDAH inhibitor in preparation of antibiotics. According to the invention, research finds that escherichia coli has Zn < 2 + >-dependent histone deacetylase-like amide hydrolase-HDAH, and the HDAH can directly influence the enzyme activity of metabolic enzyme through acetylation modification so as to regulate and control the metabolic level of bacteria; and the transcription level of metabolism-related genes can be indirectly regulated by acting on a transcription regulation factor (such as histone-like protein HU), and the metabolism of escherichia coli is synergistically regulated from two levels of post-translational modification and transcription. The invention also finds that the vorinostat (SAHA), the trichosaliocin (TSA) and the like can be combined with the HDAH and inhibit the deacetylation activity of the HDAH, are HDAH inhibitors, and inhibit the growth and migration ability of escherichia coli after being administered to the escherichia coli.

The application discloses a novel and powerful inhibitor of fatty acid amide hydrolase (FAAH), pyridinone compound (formula I) and therapeutic effect thereof in related diseases, and belongs to the technical field of biological medicine. The inhibitor can inhibit the biological activity of fatty acid amide hydrolase (FAAH), increase the physiological concentration of endogenous arachidonic acid ethanolamine, and play the roles of regulating immunity and resisting inflammation; for example, in inflammatory bowel disease, the inhibitor can significantly improve various pathological and physiological indexes of animals in an inflammatory bowel disease (IBD) model, slow down the progress of the disease, and be effectively used for treating multiple types of inflammatory bowel disease (IBD) including ulcerative colitis (UC) and Crohn's disease (CD).

The invention discloses an amide hydrolase mutant and an application in production of nicotinic acid, and provides two novel amide hydrolase mutants, which can significantly improve the production efficiency and yield of nicotinic acid, reduce the operation steps of nicotinic acid synthesis, realize one-step reaction synthesis from nicotinamide to nicotinic acid after cell disruption, break the bottleneck of the prior art, and improve the production efficiency and yield of nicotinic acid. The synthesis time is greatly shortened while the production cost of the nicotinic acid is reduced, and the production of the nicotinic acid is promoted to develop towards a more efficient, more economic and more environment-friendly direction.

The invention provides an application of URB597 in preparation of a medicine for treating chronic traumatic encephalopathy. The URB597 inhibits neuronal ferroptosis through an endocannabinoid-ACSL4 axis so as to relieve cognitive impairment after CTE. The URB597 disclosed by the invention can inhibit the activity of fatty acid amide hydrolase and increase the level of endocannabinoid AEA in the brain, and the AEA has the capability of reducing neuronal ferroptosis by directly combining and interacting with 447-site serine (Ser447) of acyl coenzyme a synthetase long-chain family member 4 (ACSL4). The effect of remarkably improving the cognitive function of the chronic traumatic encephalopathy is reflected. Aiming at the effect of the URB597, the URB597 can be applied to preparation of medicines for treating chronic traumatic encephalopathy.

The invention discloses application of amide hydrolase OtaH in degradation of ochratoxin A, and relates to the technical field of enzyme engineering and food safety biology. The amino acid sequence of the amide hydrolase OtaH is as shown in SEQ ID NO.2, and the coding gene sequence of the amide hydrolase OtaH is as shown in SEQ ID NO.1. The invention provides the application of the amide hydrolase OtaH in degradation of ochratoxin A. The invention further provides the application of the amide hydrolase OtaH in degradation of ochratoxin A. The amide hydrolase OtaH can specifically hydrolyze an amido bond of ochratoxin A (OTA) to generate almost non-toxic ochratoxin alpha, has high catalytic efficiency, can completely degrade 12.4 mu M OTA within 5 minutes in the enzyme amount of 5.0 mu g / mL, still keeps more than 50% of activity after being incubated for 1 hour at 65 DEG C, shows excellent thermal stability, can be widely applied to detoxification treatment of OTA in feed and food, and has a wide application prospect in the treatment of ochratoxin alpha, ochratoxin alpha, ochratoxin alpha, ochratoxin alpha, ochratoxin alpha, ochratoxin alpha, ochratoxin alpha, ochratoxin alpha, ochratoxin alpha, ochratoxin alpha and ochratoxin alpha. The harm of OTA to human beings and animals is reduced.

The present disclosure relates to fatty acid amide hydrolase (FAAH) modulators, inhibitors, or FAAH modulators and inhibitors and methods and uses thereof. The FAAH modulators, inhibitors, or FAAH modulators and inhibitors may be compounds having Formula I, II, III or IV. Pharmaceutical compositions comprising the FAAH modulators, inhibitors, or FAAH modulators and inhibitors are also provided.

The invention discloses an application of amide hydrolase in preparation of chiral amines. The invention provides a preparation method of a compound II, which comprises the following steps: in a solvent, in the presence of an enzyme, a compound I is subjected to a hydrolysis reaction as shown in the following formula to obtain the compound II, and the enzyme has an amino acid sequence as shown in SEQ ID NO: 1. The amide hydrolase with the amino acid sequence as shown in SEQ ID NO: 1 can selectively hydrolyze an L-configuration substrate to obtain an L-configuration amino acid, has a good conversion rate, and is wide in applicable substrate range. # imgabs0 #

The present disclosure relates to fatty acid amide hydrolase (FAAH) modulators, inhibitors, or FAAH modulators and inhibitors and methods and uses thereof. The FAAH modulators, inhibitors, or FAAH modulators and inhibitors may be compounds having Formula I, II, III or IV. Pharmaceutical compositions comprising the FAAH modulators, inhibitors, or FAAH modulators and inhibitors are also provided.

The invention discloses an amide hydrolase mutant and application of the amide hydrolase mutant in preparation of chiral gamma-amino acid drug intermediates, and belongs to the technical field of bioengineering. Wild type GsHaeWT with an amino acid sequence as shown in SEQ ID No.1 is used as a starting enzyme, further molecular modification is performed on the starting enzyme through a protein engineering means to obtain a plurality of amide hydrolase mutants with remarkably improved catalytic activity and stability, the amide hydrolase mutants and nucleic acid for coding the amide hydrolase mutants are provided, and the amide hydrolase mutants and the nucleic acid for coding the amide hydrolase mutants are used for preparing the amide hydrolase. The invention relates to a nucleic acid, a recombinant expression vector containing the nucleic acid, a recombinant expression transformant containing the expression vector, a recombinant amide hydrolase mutant catalyst and application of the amide hydrolase mutant or the recombinant amide hydrolase mutant catalyst in preparation of chiral gamma amino acid drug intermediates. The amide hydrolase mutant provided by the invention has the advantages of high protein expression quantity, high catalytic activity, good thermal stability, high product optical purity and the like, and shows excellent industrial application prospects.

The application belongs to the field of synthesis of pharmaceutical intermediates and discloses a lactam hydrolase mutant, a decarboxylase mutant and application of the lactam hydrolase mutant and the decarboxylase mutant in preparation of 5-methoxytryptamine. The lactam hydrolase mutant and the decarboxylase mutant with high activity are obtained by mutating specific positions of the lactam hydrolase and the decarboxylase. When the lactam hydrolase mutant and the decarboxylase mutant are used in preparation of 5-methoxytryptamine, the conversion rate and the yield are improved, the conditions are mild, and the use of acid and alkali reagents is reduced.

The invention discloses an amide hydrolase and an application of the amide hydrolase in degradation of ochratoxin. The invention provides a protein or a mutant thereof. The protein or the mutant thereof comprises a protein as shown in an amino acid sequence of SEQ ID NO: 1; or a protein which is obtained by mutating the 76th amino acid residue S on the amino acid sequence as shown in SEQ ID NO: 1 into E and has the same function; or a protein which is obtained by mutating the 334th amino acid residue V on the amino acid sequence as shown in SEQ ID NO: 1 into E and has the same function as the amino acid residue V on the amino acid sequence as shown in SEQ ID NO: 1; or the N end or / and C end of each protein is / are connected with a tag to obtain a protein shown as a sequence. The invention finds that the amide hydrolase ThADH and the mutant can efficiently degrade OTA (Ochratoxin A) and realize OTA detoxification.

The invention discloses a method for preparing a niraparib chiral intermediate by a chemical-enzyme method. According to the method disclosed by the invention, a compound II is taken as an initial raw material, a compound V is obtained through esterification, Michael addition and cyclization reaction, then a chiral compound VI is obtained under the action of amide hydrolase (AMD), the conversion rate and ee value can reach 99%, and finally the niraparib chiral intermediate I is obtained through reduction reaction of the compound VI. The route is simple to operate and low in cost, and can be used for industrial production.

The invention relates to the technical field of enzyme engineering, synthetic biology and microbial fermentation, in particular to an amide hydrolase mutant and application thereof. The substrate range of the amide hydrolase RsAD from the microorganism Rhodococcus sp. Is relatively narrow, and after the amino acid residue L447 in the substrate channel of the amide hydrolase RsAD is mutated into alanine, the reaction enzyme activity of the alanine on the large-volume substrate, namely indoleacetamide, is increased, so that the production efficiency of indoleacetic acid is greatly improved.

The application belongs to the technical field of biological medicine, and particularly discloses a method for preparing L-histidine from L-carnosine mother liquor. The method mainly comprises the following steps: A. vacuum concentrating the L-carnosine mother liquor into a mother liquor concentrate, adding a strong acid, a strong base or an amide hydrolase into the mother liquor concentrate to hydrolyze the peptide component into a hydrolysate mainly containing amino acids; B. removing the acid, the base or the enzyme from the hydrolysate, and then concentrating; after the concentration, sample loading column chromatography is performed; C. sequentially using purified water and gradient concentration ammonia water for elution, collecting the eluate, and using HPLC for tracking detection of the purity of L-histidine in the eluate, and collecting the eluate with the purity of L-histidine of 94% to 99%; D. combining the eluate, vacuum concentrating the eluate into a solution with the weight concentration of L-histidine of 30 to 40%, and adding simple alcohol for crystallization; E. filtering the precipitated L-histidine crystals, washing the filter cake with simple alcohol, drying by suction and drying, and then the L-histidine crystals can be obtained.

The application discloses application of an amide hydrolase OtaH in degrading ochratoxin A, and relates to the technical field of enzyme engineering and food safety biotechnology. The amino acid sequence of the amide hydrolase OtaH is shown as SEQ ID NO. 2, the coding gene sequence of the amide hydrolase OtaH is shown as SEQ ID NO. 1, and the application provides application of the amide hydrolase OtaH in degrading ochratoxin A. The amide hydrolase OtaH can specifically hydrolyze the amide bond of ochratoxin A (OTA) to generate almost non-toxic ochratoxin alpha, the enzyme has high catalytic efficiency, 5.0 mu g / mL of the enzyme amount can completely degrade 12.4 mu M OTA in 5 minutes, and more than 50% activity is still maintained after incubation at 65 DEG C for 1 hour, thereby showing excellent thermal stability. The amide hydrolase OtaH can be widely used in detoxification treatment of OTA in feed and food, and the harm of OTA to human beings and animals is reduced.

Creatinine levels may be monitored as a measure of kidney function. Conventionally, blood and / or urine tests are used for this purpose. Analyte sensors capable of monitoring creatinine in vivo may comprise: a sensor tail comprising at least a first working electrode, a creatinine-responsive active area disposed upon a surface of the first working electrode, a first membrane that is permeable to creatinine and overcoats the creatinine-responsive active area, and an oxygen scavenger located upon the sensor tail in proximity to the creatinine-responsive active area. The creatinine-responsive active area comprises a first electron transfer agent, a first polymer, and an enzyme system comprising multiple enzymes, particularly creatinine amidohydrolase, creatine amidohydrolase, and sarcosine oxidase, that are capable of acting in concert to facilitate detection of creatinine. An oxidase enzyme may serve as the oxygen scavenger, particularly glucose oxidase when detecting creatinine in fluids also containing glucose.

The present invention relates to the field of enzyme engineering technology, and in particular to an enzyme composition and its application in the preparation of allylcysteine. The present invention provides an efficient allylcysteine holoenzymatic synthesis process, wherein the overall process uses allyl benzoate and glutathione as raw materials, and directly obtains the target product through three consecutive reactions of thioester ligase catalytic coupling, metal-dependent hydrolase directional cleavage, and amidohydrolase demodification. The process can use a crude enzyme liquid system to complete the conversion in one step, or can use an immobilized enzyme to achieve the recycling of the enzyme catalyst. The coenzyme involved in the reaction can be regenerated in situ by a cascade regeneration system, further reducing material costs. Therefore, the allylcysteine production process developed by the present invention has the comprehensive advantages of a simple reaction path, mild conditions, high product purity, and easy industrial scale-up.

The present invention relates to the fields of enzyme engineering, synthetic biology and microbial fermentation technology, and in particular to an amidohydrolase mutant and its application. Rhodococcus sp. The substrate range of the amidohydrolase RsAD is relatively narrow. After the amino acid residue L447 in the substrate channel of the amidohydrolase RsAD is mutated to alanine, its reaction activity for the large-volume substrate, indoleacetamide, is increased, thereby greatly improving the production efficiency of indoleacetic acid.

The present invention relates to the technical field of molecular breeding. The present invention discloses an auxin amide hydrolase RhILL1 gene and its application in regulating the flower color of roses. The nucleotide sequence of the auxin amide hydrolase RhILL1 gene is shown in SEQ ID NO.1. The present invention effectively regulates the synthesis of anthocyanins by silencing or overexpressing the RhILL1 gene, thereby achieving precise control of the color of petals. Studies have found that silencing the RhILL1 gene causes the color of the petals to fade and the anthocyanin content to decrease; while overexpressing the RhILL1 gene prolongs the color retention time of the petals. In addition, the present invention also provides a kit and a recombinant plasmid containing the RhILL1 gene regulatory sequence, as well as corresponding gene editing and transgenic breeding methods. These achievements provide new molecular tools for rose breeding and flower color improvement, and have important ornamental value and economic significance.

The invention discloses an amide hydrolase mutant and application thereof. The invention provides a mutant protein of amidase CaADH, which is obtained according to any one of the following modes: the 67th amino acid residue G on the amino acid sequence of the amidase CaADH as shown in SEQ ID NO: 1 is mutated into D, or the 67th amino acid residue G is mutated into E, or the 67th amino acid residue G is mutated into L, and the mutant protein has the same function; experiments prove that the amide hydrolase CaADH and the mutant thereof can be used for efficiently degrading OTA and realizing OTA detoxification.

The invention discloses a novel potent inhibitor pyridone compound (formula I) of fatty acid amide hydrolase (FAAH) and a treatment effect of the novel potent inhibitor pyridone compound in related diseases, and belongs to the technical field of biological medicines. The biological activity of fatty acid amide hydrolase (FAAH) can be inhibited, the physiological concentration of endogenous arachidonic acid ethanolamine is increased, and the effects of regulating immunity, resisting inflammation and the like are achieved; taking inflammatory bowel disease as an example, the inhibitor can significantly improve various pathological and physiological indexes of an animal level in an inflammatory bowel disease (IBD) model and slow down the progress of the disease course, and is effectively used for treating various types of inflammatory bowel diseases (IBD) including ulcerative colitis (UC) and Crohn disease (CD).

The invention discloses an amide hydrolase mutant, a biological material, a screening method, a catalyst and application, and compared with wild type amide hydrolase, the amide hydrolase mutant has amino acid residue mutation at the 317th site, or amino acid residue mutation at the 317th site and amino acid residue mutation at the 63rd site, or amino acid residue mutation at the 317th site, amino acid residue mutation at the 63rd site and amino acid residue mutation at the 65th site. The catalytic efficiency of the mutant can reach 16 times as high as that of wild type amide hydrolase, the yield of R-isobutylglutaric acid monoamide generated by catalyzing isobutylglutaric imide reaches 99%, the stereoselectivity ee value is greater than 99%, the reaction condition is mild, the reaction can be carried out only at 20-45 DEG C, and the mutant has high industrial application value.