59 results about "Ester hydrolase" patented technology

The invention relates to a method used microorganism enzyme resolution DL-pantoic acid lactone to produce D-pantoic acid. It uses the D-pantoic acid lactone hydrolase strain of the fusarium, gibberella, aspergillus, penicillium, rhizopus, gliocladium, aureobasidium to ferment and culture, uses wet thallus as coarse enzyme, DL-pantoic acid lactone as substrate to produce D-pantoic acid. L- pantoic acid lactone can be reclaimed. The DL-pantoic acid lactone gained by racemization reaction can newly be used to do resolution.

The invention discloses a saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol as well as a building method of the saccharomyces cerevisiae strain, and belongs to the technical field of bioengineering. According to the building method provided by the invention, through completely knocking out an amino acid transaminase gene BAT2 and an ester hydrolase gene IAH1 in an original strain, and selecting a strong promoter PGK1 over-expression alcohol acetyltransferase I gene ATF1 at the same time, the saccharomyces cerevisiae strain with high yield of ester and low yield of higher alcohol is obtained. Compared with a parent strain, other fermentation performances of built recombinant bacteria are not affected, the total quantity of acetic acid ester is obviously increased and reaches 1303.6mg/L, wherein the content of ethyl acetate is 52 times that of the original strain, isoamyl acetate is increased to 73.7mg/L, the content of main higher alcohol is 151.8mg/L and is reduced by 61.4 percent in comparison with that of the original strain. By using the saccharomyces cerevisiae, ester yield is significantly increased while the higher alcohol yield is reduced, the higher requirements of white spirit related fields on yeast are met and the application prospect is wide.

The invention relates to laevorotation lactone specificity hydrolytic enzyme producing strain and the method used it to make chirality oxyacid. The enzyme producing is Fusarium proliferatum Nirenberg ECU2002 with storage number CGMCC 1494. The chirality oxyacid preparing method includes the following steps: using the fungous mycelium, rough enzyme extract or their immobilization derivative as biocatalyst; processing antipode selectivity hydrolysis resolution for a series of racemic chirality lactone to gain many optically active (+)-oxyacid and (+)-lactone which can be hydrolyzed into (-)-oxyacid; (+)-alpha-hydroxyl-beta, beta-dimethyl-gamma-butyric acid which are D-(+)-pantoic acid; simple acidizing to gain chirality intermediate D-(-)-pantoic acid lactone widely used in preparing feed and daily chemical engineering industry.

Methods to control organic contaminants in fibers are described. One method involves contacting the fibers with a) at least one zeolite and optionally b) detackifier, or an ester hydrolyzing enzyme, or both, for a sufficient time and in a sufficient amount to control the organic contaminants present in the fibers. This method is effective to reduce stickies in paper mill furnish formed with recycled fibers. A method for pitch control in paper mill furnish formed with virgin fibers is also provided. Resulting paper products formed from the processed fibers are also described as well as methods to make them.

The invention discloses a heterogeneous biocatalyst taking alpha-amino-acid ester hydrolase as basis, a preparation method thereof and application in synthesizing amino beta-lactams. The activity of catalyst synthetase is 2,200 U/g by dry weight, and the content of protein is 0.95 g/g by dry weight; and xanthomonas rubrilineans cell biomass is processed in an organic solvent under a pH gradient through low temperature effect to extract enzyme, and enzyme aggregates are deposited and cross-linked to obtain the hydrolase. The alpha-amino-acid ester hydrolase as a catalyst can be synthesized into amino penicillin and amino cephalosporin drugs through acylating a beta-lactam compound by D-phenylglycine methyl ester derivatives in water or in a mixed medium of water and an organic solvent.

The invention relates to the field of biochemical engineering, in particular to a novel process for preparing L-amino acid and D-amino acid by direct biocatalysis and hydrolysis resolution for DL-N-acetyl amino acid esters. DL-N-acetyl amino acid esters is used as substrate, enzyme including amide hydrolase and ester hydrolase is used as catalyst, wherein the enzyme is achieved by fermenting aspergillus oryzae Lb3085, de-ionized water is utilized as solvent and added in an enzyme catalysis reactor, and L-N-acetyl amino acid esters of the substrate is hydrolyzed into L-amino acid, D-N-acetyl amino acid esters of the substrate is hydrolyzed into D-N-acetyl amino acid, thereby resolution is realized. The resolution technique of 'one bacterium two enzymes' can efficiently reduce operating intermediate process, and achieve purposes of convenience and cleaning.

The invention provides esterolytic enzyme from bacillus megaterium, a coding gene of the esterolytic enzyme, a carrier containing the coding gene, an engineering bacterium and application of the coding gene. The amino acid sequence of the esterolytic enzyme is shown as SEQ ID NO. 2; the nucleotide sequence of the encoding gene is shown as SEQ ID NO.1. The recombined esterolytic enzyme provided by the invention can be used for synthesizing a stereoselective catalytic chiral compound and has high catalytic activity and stereoselectivity.

The invention provides tertiary alcohol ester hydrolase, a gene for encoding the same, a carrier containing the encoding gene, engineering bacteria and application thereof. The amino acid sequence of the tertiary alcohol ester hydrolase is shown as SEQ ID No.2, and the amino acid sequence of the encoding gene is shown as SEQ ID No.1. The tertiary alcohol ester hydrolase is high in short-carbon chain ester hydrolytic activity; and the enzymatic activity for catalyzing hydrolysis of p-nitrophenol butyrate in a Tri-HCl buffer solution with pH of 8.5 at the temperature of 50 DEG C can reach 3,010 U/mg. The esterase is high in strong organic solvent resistance stability and activity improving characteristic; and after the esterase is incubated in strong organic solvents such as normal hexane, toluene and dimethylbenzene, of which the partition coefficient Log P is greater than 3, the enzymatic activity still can be improved by 51 percent, 44 percent and 49 percent respectively. Moreover, the esterase has tertiary alcohol ester hydrolytic activity, so that the esterase can be widely used for hydrolyzing different tertiary alcohol ester substrates. The esterase, the encoding gene, the carrier, and the recombinant bacteria can be widely applied to ester catalysis of synthesis and resolution of chiral compounds related to tertiary alcohol substrates in medicine industry, food industry, chemical industry and the like.

Methods are described for detecting chemical warfare agents that contain organophosphorus compounds. Compositions containing either (1) a sufficient amount of phosphonate ester hydrolase with an alkali agent or (2) a sufficient amount of phosphonate ester hydrolase, a sufficient amount of organophosphorus hydrolase and a sufficient amount of a organophosphorus acid anhydrolase, may be utilized to test for organophosphorus compound-containing chemical warfare agents. These enzymes will react with such chemical warfare agents to produce degradation products, specifically phosphonate esters.

The invention provides high-R-isomer-stereoselectivity esterolytic enzyme, an encoding gene of the esterolytic enzyme, a carrier containing the encoding gene, an engineering bacterium and application of the encoding gene. An amino acid sequence of the esterolytic enzyme is shown as SEQ ID No.2, and a nucleotide sequence of the encoding gene is shown as SEQ ID No.1. The esterolytic enzyme provided by the invention has higher catalysis activity and stereoselectivity, and can be used for preparing an optically pure chiral compound, particularly a levetiracetam intermediate, namely alpha-ethyl-2-oxygen-1-pyrrolidine methyl acetate.

The invention discloses an application of zearalenone lactone hydrolase RmZHD in degradation of macrolide antibiotics. Research finds that the zearalenone hydrolase RmZHD can efficiently degrade macrolide antibiotics, and can be used for reducing antibiotic pollution in the environment. The invention also provides a recombinase RmZHD-rHF containing the zearalenone hydrolase RmZHD. The activity andexpression quantity of the enzyme of RmZHD can be further improved, and the application prospect is great.

The invention discloses a zearalenone lactone hydrolase mutant S162P with improved thermal stability and application thereof, and belongs to the technical field of enzyme engineering. According to the invention, zearalenone lactone hydrolase derived from a microorganism GliocladiumroseumMA918 is adopted as a parent, and a gene mutation technology is adopted to mutate serine (S) at the 162 site into proline (P) so as to obtain the single-site mutant S162P. Compared with a wild enzyme, on the basis that the original catalytic activity is not obviously changed, the thermal stability of the mutant enzyme is obviously improved: after the mutant enzyme is kept at 50 DEG C for 2 minutes, the residual enzyme activity is improved to be 1.52 times of that of the wild enzyme; and after heat preservation is conducted for 10 min and 20 min at the temperature of 48 DEG C respectively, about 80% and 58% residual enzyme activities are still remained, and the half-life period at the temperature of 48 DEG C is also increased by 3.6 times that of the wild enzyme. The invention lays a certain foundation for industrial application of the ZEN lactone hydrolase.

The invention relates to a urine protein marker of ovarian cancer and a diagnosis application thereof, in particular to a urine protein marker obtained by using in-situ injection of ovarian cancer cells in a rat ovary and mass spectrum analysis, and application in the diagnosis and disease course monitoring of human ovarian cancer. The urine protein marker is selected from cystathionine-gamma-lyase, a Rab GDP dissociation inhibitor beta, an ester hydrolase C11orf54 homolog, neutral and alkaline amino acid translocator rBAT, apolipoprotein A-1, OX-2 membrane glycoprotein, charged multivesicularbody protein 5, beta-amino hexosaminidase subunit beta, monocyte differentiation antigen CD14, glyceraldehydes-3-phosphate dehydrogenase, cadherin 1, gastricsin, a serine protease inhibitor A3M, alpha-1-asialoglycoprotein and the like. Differential protein obtained from the marker provides a simpler, quicker and non-invasive choice for early diagnosis and disease course monitoring of ovarian cancer.

The invention provides a pantolactone hydrolase mutant strain and application thereof. Specifically, the invention provides a mutant pantolactone hydrolase with significantly improved enzyme activity and a host cell for expressing the mutant pantolactone hydrolase. The invention further provides an enzyme preparation containing the mutant pantolactone hydrolase, D-pantolactone in a substrate DL-pantolactone can be completely hydrolyzed into D-pantoic acid, and the mutant pantolactone hydrolase is high in conversion rate, stable in property, capable of being repeatedly used and suitable for industrial production.

The invention discloses a high-purity EPA glyceride and DHA glyceride preparation method. The method includes steps: (1) performing fish oil enzyme catalytic hydrolysis, to be specific, adding water, fish oil and glyceryl ester hydrolase into a reactor, enabling reactants to be in an emulsified state under the condition of stirring, and adjusting pH, wherein final reaction products are fatty acid and glycerin aqueous solution; (2) separating and purifying fish oil fatty oil by a film evaporator and a rectifying tower to obtain high-purity EPA and DHA; (3) synthesizing EPA glyceride from EPA and glycerin through lipase; (4) synthesizing DHA glyceride from DHA and glycerin through lipase. The method is free of chemicals such as ethyl alcohol and urea, and a production process is simple; a multistage molecular distillation separation method which fails to realize large-scale production is substituted by a rectifying tower separation method which is easy to implement large-scale production, so that production efficiency is improved, and industrial environmental pollution is abated.

The invention discloses a Bacillus sp. Code with R-isomer stereoselectivity of esterase gene BSEST, which provides BSEST sequence and amino acid sequence, the expression carrier Pbsest-pet with BSEST and the gene engineering bacterial with the carrier(E. coli BL21(DE3)/pBSEST-PET CGMCC No. 1648), and also provides the recombinant method of the esterase. The recombinant esterase can apply to the synthesis of stereoselectivity catalytic chirality compounds(R shape).

The invention discloses a recombinant bacterium of lipase expressing R configuration selectivity and application thereof. A lipase gene with R configuration selectivity is inserted into a multiple cloning site of a pET28a plasmid to obtain a recombinant plasmid; the recombinant plasmid is transformed into colibacillus SHuffle T7 to obtain the recombinant bacterium; the gene sequence of the lipasegene with the R configuration selectivity is a gene sequence of the following 1) or 2), wherein 1) is a DNA sequence shown as SEQ ID NO.2 in a sequence table; 2) is a DNA sequence with 90% homology orabove with the DNA sequence defined in 1) and used for encoding protein with esterolysis enzymatic activity. The recombinant lipase has higher R-configuration steric selectivity and catalytic activity, more than 85% of the enzyme activity is reserved under a condition with higher temperature, and the recombinant lipase can be used for splitting racemization 2-aryl-methyl propionate substances, and obtain optical pure S-type 2-aryl-methyl propionate substances.

The invention discloses a D-pantolactone hydrolase producing bacteria in the technical field of bioengineering, and relates to a Thielavia sp. Zmu20201 serving as a microorganism, and the preservationnumber of the strain is CCTCC: M2020062. Meanwhile, the invention further discloses the D-pantolactone hydrolase from the Thielavia sp. Zmu20201 strain and an engineering bacterium strain of the D-pantolactone hydrolase, wherein the amino acid sequence number of the D-pantolactone hydrolase is as shown in SEQ ID NO.2. In addition, the invention also discloses an application of the Thielavia sp. Zmu20201 fermentation thallus or the lactone hydrolase gene engineering strain fermentation thallus as a biocatalyst in resolution of D, L-pantolactone to prepare optically pure D-pantoic acid.