97 results about "Acid Esterase" patented technology

The invention discloses a thermostable carboxylesterase gene, coding protein and application thereof, the esterase has amino acid sequence shown in SEQ ID NO:1. Compared with the prior art, the esterase containing EstW is thermostable esterase obtained from streptomycete in known mesophilic bacteria, can be widely applied to production of non-steroidal anti-inflammatory drugs or ferulic acid.

The invention provides a specific fluorescence probe substrates of human carboxylesterase 2 (CES2) and application thereof. The specific probe substrate is a benzoateb compound of a C4 hydroxyl naphthalimide, and is applicable to determine the enzyme activity of CES2 in a biological system. The CES2 enzyme activity determination flow comprises: selecting a hydrolysis benzoyl-removal reaction of the benzoate compound of the C4 hydroxyl naphthalimide as a probe reaction, and quantitatively determining the generation amount of a hydrolysis metabolite of the compound in a unit time, so as to determine the enzyme activity of CES2 in all biological samples, cells, bodies and integral organs. The probe is applicable to quantitative assessment of CES2 enzyme activity in biological samples of different species and different individual sources, and quantitative determination on CES2 enzyme activity in different sources of animal tissue cell culture fluids and cell preparation substances, so that the probe is expected to help to realize assessment on medicine disposal capability of important drug metablic enzyme CES2. Additionally, the probe also is applicable as an inhibitor for rapidly screening CES2 in vitro by means of the probe reaction.

The invention discloses a chromogenic medium for detecting salmonella, which belongs to the fields of food safety and clinical microbiological detection. The culture medium contains agar, peptone, beef extract powder, sodium chloride, a surfactant, cholate, an enzyme inducer, an octoate enzyme chromogenic substrate, a beta-galactosidase chromogenic substrate, a hexosaminidase chromogenic substrate and robiocina. The chromogenic medium disclosed by the invention is used for detecting salmonella, has high detection sensitivity and high specificity, and can be used for initially identifying strains directly according to the color of a colony; the chromogenic medium has high operability, and suitable for treating large-reflux samples, and can be used for comprehensively, systematically and accurately detecting and initially identifying salmonella in food production, clinical disease diagnosis and the environment; and a new way is provided for rapid detection of microorganisms.

Compounds of formula (I) are inhibitors of P13 kinase activity, and useful in treatment of, inter alia, autoimmune, inflammatory and proliferative diseases: wherein: s is 0 or 1; U is hydrogen or halogen; X is —(C═O), an optionally substituted divalent phenylene, pyridinylene, pyrimidinylene, or pyrazinylene radical, or a bond; P is optionally substituted C₁-C₆ alkyl and Z is —(CH₂)_Z—X₁-L₁-NHCHR₁R₂; or Z is optionally substituted C₁-C₆ alkyl and P is —(CH₂)_Z—X₁-L₁-NHCHR₁R₂; R₁ is a carboxylic acid group (—COOH), or an ester group which is hydrolysable by one or more intracellular carboxylesterase enzymes to a carboxylic acid group; R₂ is the side chain of a natural or non-natural alpha amino acid; X₁ is (i) a bond; —NR₄C(═O)NR₅— or —NR₄S(═O)₂—; or except when X is —(C═O)— (ii) —C(═O)—, —S(═O)₂—, or —S(═O)₂NR₄— wherein R₄ and R₅ are independently hydrogen or optionally substituted C₁-C₆ alkyl; and z and L1 are as defined in the specification.

The invention provides feruloyl esterase and a preparing method and application thereof. A feruloyl esterase gene coming from a soil macro gene library have the nucleotide sequence and amino acid sequence shown in SEQ ID NO.1 and SEQ ID NO.2. The gene contains a tetrapeptide SXXK sequence motif which is rarely seen, and after the esterase gene is inserted into plasmid pET28a(+), the gene is transformed into escherichia coli BL21(DE3) to achieve heterogeneous expression. The molecular weight of purified recombinase (DLFae4) is 38.3 kDa. Besides, it is put forward for the first time that novel feruloyl esterase can hydrolyze penbritin, penicillin, cefazolin and other lactam antibiotics. As is shown by site-directed mutagenesis experiments, a catalysis triplet of DLFae4 is composed of serine(S11), histidine (H74) and aspartic acid (D302), and the mutation of any of serine (S11), histidine (H74) and aspartic acid (D302) can cause loss of the catalysis capability of DLFae4. DLFae4 has a high hydrolytic activity on methyl ferulate and has good heat stability. In the presence of cellulase, DLFae4 can obviously increase the amount of ferulic acid released from destarched wheat bran. Due to peculiar activities and enzymatic characteristics of novel feruloyl esterase, novel feruloyl esterase can be applied to feed, paper making, food, pharmacy and other fields.

The invention discloses functional yellow wine enriched in ferulic acid and a production method of the functional yellow wine, belonging to the technical field of food brewing. The aspergillus niger CCTCC NO:M 2015202 feruloyl esterase, amylase and cellulase are high in activity, when the enzymes are added into the yellow wine fermentation process, normal fermentation of the yellow wine is not influenced, saccharifying of raw materials in the yellow wine production process is accelerated, the yellow wine production and fermentation cycle is shortened, and the original flavor and taste of the yellow wine can be maintained. The method disclosed by the invention comprises the following steps: by taking sticky rice as a raw material, inoculating aspergillus niger strain spore suspension containing high-yield feruloyl esterase during jar dropping, adding wheat bran with high ferulic acid content, performing primary fermentation, secondary fermentation, pressing, boiling wine, filling, sealing the jar and the like. The yellow wine produced by the method disclosed by the invention is enriched in ferulic acid and has the effects of resisting oxidation, reducing blood pressure, resisting thrombus, reducing blood fat, resisting bacteria, diminishing inflammation, preventing cancers, protecting the liver, treating the diabetes and the like. Moreover, the production process is energy saving and environmentally friendly, and the additional value of byproducts of wheat production is improved.

The invention provides a feruloyl esterase gene. The feruloyl esterase gene is originated from microbes in the rumen of a Chinese yak and has a nucleotide sequence as shown in SEQ ID No. 1, wherein the nucleotide sequence codes an amino acid sequence as shown in SEQ ID No. 2. The invention also provides a recombinant Kluyveromyces marxianus expression vector containing the gene, a genetically engineered strain, a preparation method for feruloyl esterase and application of prepared feruloyl esterase. Feruloyl esterase is subjected recombinant expression in a Kluyveromyces marxianus expression system; and after high-density fermentation for 48 h, intracellular and extracellular feruloyl esterase vitalities are 686.35 U/mL and 346.34 U/mL, respectively. Feruloyl esterase having undergone recombinant expression by Kluyveromyces marxianus can release ferulic acid in corn bran. Feruloyl esterase prepared in the invention can be applied to food processing, feed addition, biomedicine, biotransformation of cellulosic substances, bioenergy and other fields.

The invention discloses novel feruloyl esterase, a mutant and an application thereof, and belongs to the field of bioengineering. The method comprises the following steps: firstly, successfully screening out a feruloyl esterase gene from anaerobic fungi Neocallimatix sp. by utilizing a genomics technology; according to the preference of a prokaryotic system codon, the GC content of a feruloyl esterase gene and the secondary structure of mRNA, and codon optimization is carried out on the feruloyl esterase gene to obtain a novel feruloyl esterase gene sequence. The enzyme has an esterase regionand a glucoside hydrolase 11 region, site-specific mutagenesis is carried out on the two regions; the obtained mutant gene is also expressed in escherichia coli; separation and purification results show that the novel feruloyl esterase and the mutant thereof are successfully expressed in escherichia coli, and the esterase activity of the mutant is far lower than that of a non-mutant, so that the strain source of the feruloyl esterase is enriched, and the feruloyl esterase has theoretical guiding significance in the aspects of improving the digestibility of feed fibers and improving the production performance of animals.

Compounds of formula (I) are inhibitors of histone deacetylase activity, and are useful in the treatment of, for example, cancers, wherein R₁ is a carboxylic acid group (—COOH), or an ester group which is hydrolysable by one or more intracellular carboxyesterase enzymes to a carboxylic acid group; R₂ is the side chain of a natural or non-natural alpha amino acid; Y is a bond, C(═O)—, —S(═O)₂—, —C(—O)O—, —C(O)NR₃—, —C(═S)—NR₃, —C(═NH)NR₃ or —S(═O)₂NR³— wherein R₃ is hydrogen or optionally substituted C₁-C₆ alkyl; L is a divalent radical of formula -(Alk¹)_m(O)_n(Alk²)_p— wherein m, n and p are independently 0 or 1, Q is (i) an optionally substituted divalent mono- or bicyclic carbocyclic or heterocyclic radical having 5-13 ring members, or (ii), in the case where both m and p are 0, a divalent radical of formula —X²-Q¹- or -Q¹-X²— wherein X² is —O—, S— or NR^A— wherein R^A is hydrogen or optionally substituted C₁-C₃ alkyl, and Q¹ is an optionally substituted divalent mono- or bicyclic carbocyclic or hetero-cyclic radical having 5-13 ring members, AIk¹ and AIk² independently represent optionally substituted divalent C₃-C₇ cycloalkyl radicals, or optionally substituted straight or branched, C₁-C₆ alkylene, C₂-C₆ alkenylene, or C₂-C₆ alkynylene radicals which may optionally contain or terminate in an ether (—O—), thioether (—S—) or amino (—NR^A-) link wherein R^A is hydrogen or optionally substituted C₁-C₃ alkyl; X represents a bond; —C(═O); or —S(═O)₂—; —NR₄C(═O)—, —C(═O)NR₄—, —NR₄C(═O)NR₅—, —NR₄S(═O)₂—, or —S(═O)₂NR₄— wherein R⁴ and R₅ are independently hydrogen or optionally substituted C₁-C₆ alkyl; z is 0 or 1; A represents an optionally substituted mono-, bi- or tri-cyclic carbocyclic or heterocyclic ring system wherein the radicals R₁R₂NH—Y-L¹-X¹—[CH₂]_z— and HONHCO-[LINKER]- are attached different ring atoms; and -[Linker]- represents a divalent linker radical linking a ring atom in A with the hydroxamic acid group CONHOH, the length of the linker radical, from the terminal atom linked to the ring atom of A to the terminal atom linked to the hydroxamic acid group, is equivalent to that of an unbranched saturated hydrocarbon chain of from 3-10 carbon atoms.

The invention relates to a method for preparing clevidipine butyrate and also relates to a method for preparing 4-(2,3-dichlorophenyl)-1,4-dihydro-2,6-dimethyl-methoxycarbonyl-3-picolinic acid methyl ester (an intermediate I) and 4-(2,3-dichlorophenyl)-1,4-dihydro-2,6-dimethyl-methoxycarbonyl-3-pyridine carboxylic acid (an intermediate II) which are used as intermediates of the clevidipine butyrate. The intermediate I is prepared by using 2,3-dichlorobenzaldehyde, ammonium acetate and methyl acetoacetate as raw materials and performing a reaction by using ultrasound. The intermediate II is prepared by carrying out enzyme hydrolysis on the intermediate I. When the clevidipine butyrate is prepared by using the method disclosed by the invention, ammonia water is not used as the raw material, so that no irritant gas is generated. Moreover, the cyclizative condensation is carried out by using ultrasonic waves and the hydrolysis is carried out by using carboxy lesterase, so that the reaction time can be shortened and the reaction yield is increased.

Methods and kits are provided for detecting polymorphisms in carboxylesterase-1 (CES1). Several single nucleotide polymorphisms (SNPs) in CES1 in humans, and methods for detecting the same, are provided (e.g., Gly143Glu, 12754T>del). Results indicate that the Gly143Glu (9486G>A) polymorphism has an allelic frequency of 1.5% in the Caucasian population. Polymorphisms of the present invention may alter the function of the carboxylesterase-1 enzyme (hCES1). Thus, the methods and kits of the present invention may be used to personalize a therapy and/or avoid adverse consequences of altered metabolism of a therapeutic or compound (e.g., enalapril, methylphenidate, etc.) which may result due to a CES1 polymorphism. In addition, recombinant cells lines overexpressing wild-type CES1 or expressing CES1 mutants are provided. Such cell lines may be used to assess the effects of candidate compounds on CES1, and the action of CES1 on these candidate compounds.

The present invention relates to active antitumor matter and its preparation process and use, and belongs to the field of microbiological medicine technology. The active matter is prepared through convenient process with Moinascus ruber 3081 of preservation number CGMCC No. 1319. The test seed is cultured in conventional malt juice culture medijm at 20-30 deg.c for 4-8 days; and the solid fermentation culture medium consists of notoginseng hairy rootlet powder 20-30 wt% and water 70-80 wt% in natural pH. Through further solid culture at 20-30 deg.c for 15-20 days, alcohol leaching with alcohol solution for 2 days, decompression concentrating and evaporating out solvent, the active matter is obtained. The active matter has high inhibiting rate to human leukaemia cell strain K562, human lung cancer strain A549, oncogene Raf1 and c-Myc latent target tyrosine phosphoesterase cdc25a and cdc25b, so that it may be used in preparing antitumor preparation.

The invention discloses a method for producing feruloyl esterase and belongs to the technical field of bio-enzyme preparation production. The method comprises the following steps: fermenting by utilizing a genetic engineering strain of the constitutive expressed feruloyl esterase, and by taking the starch saccharification liquid, the maize slurry and the ammonia water as main raw materials, preparing the feruloyl esterase, continuously adding a fermentation culture medium and discharging part of the fermenting fluid in bathes during the fermentation engineering, continuously extending the fermenting period under a no-sundry bacterium contamination condition, and filtering by a plate frame filter, and performing ultrafiltration concentration and spray drying on the fermenting fluid to obtain the high-activity feruloyl esterase preparation. The specific steps are as follows: (1) strain preparation; (2) culture expansion; (3) submerged fermentation and feruloyl esterase production; and (4) feruloyl esterase purification. By adopting the method under the conditions of no serious sundry bacterium contamination and no obvious reduction of the enzyme activity in unit volume of the fermenting fluid, the feruloyl esterase can be continuously fermented and produced, the production period can be shortened and the production cost can be reduced.

The invention discloses a hydroxyapatite-based electrochemical probe and a construction method thereof, and a method for determining the activity and the inhibition property of BACE1. According to the present invention, an A[beta] antibody and alkaline phosphatase are co-modified on a hydroxyapatite matrix to form the hydroxyapatite-based electrochemical probe; the preparation method comprises: sequentially placing hydroxyapatite nanoparticles in a polyethylenimine solution and carrying out a reaction, placing in a glutaraldehyde solution and carrying out a reaction, and placing in a solution containing an A[beta] antibody and alkaline phosphatase and carrying out a reaction so as to obtain the probe; and the method for determining the activity and the inhibition property of BACE1 by using the hydroxyapatite-based electrochemical probe has advantages of simpleness, rapidness, high sensitivity and wide detection range, and can be widely promoted and applied.

The invention provides a Kluyveromyces marxianus recombinant strain capable of being used for preparing feruloyl esterase AnfaeA. The recombinant strain is constructed by cloning an aspergillus nigerferuloyl esterase AnfaeA gene with an optimized sequence to an expression vector and converting a Kluyveromyces marxianus host strain, and the yield of the feruloyl esterase obtained by high-density fermentation of the recombinant strain reaches 120000 U/ml. The invention also provides a method for preparing ferulic acid by hydrolyzing corncob powder with the feruloyl esterase. The feruloyl esterase AnfaeA prepared from the recombinant strain and xylanase have a synergistic effect, so that the corncob powder can be efficiently hydrolyzed to obtain the ferulic acid. The feruloyl esterase provided by the invention can be used for preparing phenolic substances such as ferulic acid, p-coumaric acid and the like through biodegradation of agricultural byproducts such as corncobs, rice bran, cornbran and the like, so that the feruloyl esterase has a wide application value in the fields of foods, papermaking, feeds, medicines and the like. Meanwhile, the preparation method is simple in process, high in yield and very suitable for large-scale production.

The invention relates to application of ferulic acid and oligosaccharide preparation by producing enzymes from Salicornia bigelovii straws fermented by microorganism. The microorganism in the invention refers to Aureobasidium pullulans; the Salicornia bigelovii straws are dried and crushed to a certain particle diameter, and then a mixed enzyme preparation of xylanase and ferulic acid esterase is generated after the metabolism of the Aureobasidium pullulans; the cell walls of the Salicornia bigelovii straws are degraded under the synergistic effect of the mixed enzyme so as to obtain physiologically active substances, such as the ferulic acid and oligosaccharide. The invention has the effects of oxidation resistance, free radical elimination, thrombus resistance, blood fat reduction, coronary heart disease prevention and cure, bacteria resistance, inflammation elimination, mutation resistance and cancer prevention, can be widely applied in industries of medication, food and cosmetics, and can prominently increase the economic value of the straws, i.e., by-products of Salicornia bigelovii.

The invention discloses a special complex enzyme system for kelp comprehensive utilization and application thereof. The complex enzyme system comprises an enzyme A and an enzyme B, wherein the enzyme A comprises any four or more than four kinds of materials from cellulose, hemicellulase, pectinase, amylase, xylanase, alkaline cellulose, pentosanase, glucanase and feruloyl esterase; the enzyme B comprises one or any two kinds of materials from alkaline protease, neutral protease, papain, trypsin, acid proteinase and pepsin. The complex enzyme system has the prominent beneficial effects that after the complex enzyme system is used, four kinds of products of fucoidin, alginic acid, mannitol and functional dietary fiber can be simultaneously extracted from kelps; the extraction ratio of four kinds of active ingredients reaches more than 90 percent (through being metered by the kelp dry weight); the comprehensive utilization rate of kelp resources is obviously improved; the product additional values of kelp processing industry are greatly increased; the technological level of relative products is improved.

The invention relates to a tanshinone I derivative, a preparation method and application thereof. The compound is a furan ring 2-substituted amide methylene modifier in the structural formula of tanshinone I, tanshinone I serves as the raw material, and the novel tanshinone derivative is prepared through a chemical synthesis method and can be used for treating cancers, serving as a tumor multi-drug resistance reversing agent, assistant medicine and a CYP1 family specific inhibitor, treating cardiovascular and cerebrovascular diseases, serving as a carboxylesterase inhibitor, treating androgen related diseases and serving as antibacterial.