170 results about "Enzyme synthesis" patented technology

The present invention provides isolated polynucleotides and isolated polypeptides. The polypeptides of the present invention have isoprenoid wax ester synthase activity or isoprenoid acyl CoA-synthetase activity. The present invention also includes methods of using the polynucleotides and polypeptides of the present invention. For instance, the methods include producing biodiesel and producing wax esters.

The present invention provides isolated polynucleotides and isolated polypeptides. The polypeptides of the present invention have isoprenoid wax ester synthase activity or isoprenoid acyl CoA-synthetase activity. The present invention also includes methods of using the polynucleotides and polypeptides of the present invention. For instance, the methods include producing biodiesel and producing wax esters.

Disclosed are novel acyl-CoA synthetases and novel acyltransferases, nucleic acid molecules encoding the same, recombinant nucleic acid molecules and recombinant host cells comprising such nucleic acid molecules, genetically modified organisms (microorganisms and plants) comprising the same, and methods of making and using the same. Also disclosed are genetically modified organisms (e.g., plants, microorganisms) that have been genetically modified to express a PKS-like system for the production of PUFAs (a PUFA PKS system or PUFA synthase), wherein the organisms have been modified to express an acyl-CoA synthetase, to express an acyl transferase, to delete or inactivate a fatty acid synthase (FAS) expressed by the organism, to reduce competition for malonyl CoA with the PUFA synthase or toincrease the level of malonyl CoA in the organism, and in one aspect, to inhibit KASII or KASIII. Additional modifications, and methods to make and use such organisms, in addition to PUFAs and oils o btained from such organisms, are disclosed, alone with various products including such PUFAs and oils.

The invention discloses a method for constructing an ALA production strain, that is, enhancing the activity of related enzymes promoting the synthesis of oxaloacetate in the 5-aminolevulinic acid production strain or introducing exogenous related enzymes promoting the synthesis of oxaloacetate , such as phosphoenolpyruvate carboxylase or pyruvate carboxylase, and / or weaken the succinyl-CoA downstream metabolic pathway-related enzymes in the strain, such as succinyl-CoA synthetase or succinate dehydrogenase active. The invention also discloses an ALA high-yield strain constructed by the method and a method for preparing ALA by using the strain. The bacterial strain of the invention can produce ALA efficiently, at low cost and with low pollution without adding exogenous succinic acid.

The invention relates to a gene, mutant plasmid and engineering bacteria which have improved synthesis performance to penicillin G acylase and are obtained by a gene site-directed mutagenesis method, and mutant enzyme can also be obtained with improved synthesis performance to penicillin G acylase by fermenting and purifying the engineering bacteria. Two enzymes Kpn I and Pst I are firstly used for cutting pUC18 by the invention, then T4 polymerase is adopted to make the ends blunt, and pZ01 is obtained through self-linkage; the enzyme of EcoR I is used for cutting pZ01, and then connected with pEES102 that is also cut by the enzyme of EcoR I, thereby obtaining the recombinant plasmid pY020; the pY020 is adopted as a template plasmid, and TaKaRa MuTanBEST Kit is utilized for conducting the site-directed mutagenesis to B.megaterium PGA, thereby obtaining the mutant plasmid with improved synthesis performance to the penicillin G acylase. The mutant plasmid is transformed to bacillus subtilis to obtain the required engineering bacteria. The engineering bacteria are amplified and fermented, and the mutant enzyme with improved maximum conversion rate of 7-ADCA and the ratio of synthetic product / hydrolysate can be obtained after the engineering bacteria are purified.

The invention relates to the technical field of biology, and discloses a coenzyme-Q10-production engineered bacteria construction method, engineered bacteria, and an application thereof. The method comprises the steps that: a, total genomic DNA is extracted from Rhodobacter sphaeroides cacterial liquid; b, UbiG gene is obtained by amplification with a polymerase chain reaction; c, the amplified UbiG gene is connected with broad-host plasmid, such that recombinant vector is constructed; d, the recombinant vector is transferred to Escherichia coli S17-1; and e, the Escherichia coli S17-1 is subjected to conjugal transfer with the Rhodobacter sphaeroides, such that the engineered bacteria are obtained. According to the method provided by the invention for improving coenzyme Q10 yield through regulating Rhodobacter sphaeroides aromatic ring modification pathway, the operation is simple, and coenzyme Q10 synthesis capacity can be improved by higher than 30%. The method is suitable for coenzyme Q10 large-scale industrial production.

A method for producing lacto-N-biose I and galacto-N-biose inexpensively and conveniently is provided. The method for producing lacto-N-biose I or galacto-N-biose, characterized in that the method comprises causing: (i) a combination of a carbohydrate raw material with an enzyme that catalyzes phosphorolysis of the carbohydrate raw material to give a-glucose-1-phosphate; and (ii) a combination of an enzyme that converts α-glucose-1-phosphate to UDP-glucose and an enzyme that converts UDP-galactose to galactose-1-phosphate with their cofactors, and / or a combination of an enzyme (UDP-Gly synthase) that converts α-glucose-1-phosphate and UDP-galactose to UDP-glucose and α-galactose-1-phosphate, respectively, with its cofactor(s) to act in the presence of N-acetylglucosamine or N-acetylgalactosamine, phosphoric acid, lacto-N-biose phosphorylase (EC 2.4.1.211), and UDP-glucose-4-epimerase (EC 5.1.3.2).

The invention relates to a maltose inducible trehalose synthase synthesis engineering bacterium, a method for preparing the same and application. The maltose inducible trehalose synthase synthesis engineering bacterium is characterized in that maltose inducible promoters are inserted in the fronts of BamHI cleavage sites of PHT01 plasmids of recombinant plasmid vectors instead of Pgrac promoters on the PHT01 plasmids, expression genes of Tat type signal peptides are inserted in the fronts of the BamHI cleavage sites, and expression genes of trehalose synthase are inserted in the rears of the BamHI cleavage sites. The maltose inducible trehalose synthase synthesis engineering bacterium, the method and the application have the advantage that expression effects realized after the maltose inducible promoters and the trehalose synthase are fused with one another are obviously superior to other inducible expression effects.

The present invention provides a biodegradable article formed from enzyme-synthesized amylose using phosphorylase, wherein the enzyme-synthesized amylose is composed of glucose monomers exclusively bonded by alpha-1,4-glucosidic likage and has a weight average molecular weight of not less than 100 kDa, preferable not less than 600 kDa. The enzyme-synthesized amylose employed in the present invention has a molecular weight distribution (Mw / Mn) of not more than 1.25 and can be chemically modified if desired. The biodegradable article of the present invention can be produced by (a) the enzyme-synthesized amylose and / or its modified one, or a combination of the amylose (a) and another polymer material (b).

The invention relates to the technical field of bilirubin production methods and discloses a method for synthesizing bilirubin by utilizing an immobilized enzyme. The method comprises the following steps of: a) respectively cloning haem oxygenase genes and biliverdin reductase genes; b) respectively expressing the recombinant haem oxygenase and biliverdin reductase in escherichia coli; c) extracting a crude extract or pure enzyme of haem oxygenase and a crude extract or pure enzyme of biliverdin reductase; d) immobilizing the crude extract or pure enzyme of haem oxygenase and the crude extract or pure enzyme of biliverdin reductase; and e) preparing bilirubin in the assistance of an oxidized coenzyme II by catalyzing with the immobilized haem oxygenase and the immobilized biliverdin reductase and taking haem and oxygen as substrates.

The invention discloses a method of synthesizing diacylglycerol. The method is characterized in that a fatty acid donor, glycerol, partial glyceride lipase and monoglyceride lipase are mixed with water, an esterification reaction is conducted for 8-24 h, separation and purification are further conducted, and then diacylglycerol is obtained. According to the method of synthesizing diacylglycerol, the monoglyceride lipase is utilized for promoting the reaction efficiency of the partial glyceride lipase during the esterification reaction to increase the speed of diacylglycerol synthesis; comparedwith a method using one enzyme, the method of synthesizing diacylglycerol shortens reaction time by 50% or above; and after the esterification reaction, the yield of diacylglycerol reaches 45.50% orabove, and basically no triglyceride is generated in products, so that after molecular distillation and purification, the content of DAG reaches up to 98% or above. The method of synthesizing diacylglycerol is a synthesis method which is rapid, efficient, environmentally friendly and convenient for industrial production.

The invention discloses construction and application of Escherichia coli engineering bacterium for immobilizing CO2 and producing malic acid, and belongs to the field of fermentation. The engineeringbacterium is a strain obtained by carrying out gene engineering modification on Escherichia coli MG1655. The genetic engineering transformation is as follows: 1, knocking out a fumarate reductase gene, a fumarase gene, a lactic dehydrogenase gene and an ethanol dehydrogenase gene, and carrying out free over-expression on formate dehydrogenase, acetyl coenzyme A synthetase, acylated acetaldehyde dehydrogenase, formaldehyde lyase, dihydroxyacetone kinase, malic enzyme and phosphorous acid oxidordeuctase so as to obtain a strain GH0407. The strain is used for fermentation production of malic acid, CO2 and glucose are used as co-substrates for anaerobic fermentation for 72 h, the malic acid yield reaches 39 g / L, the yield is 1.53 mol / mol, and malic acid is not accumulated in an original starting strain.

The invention discloses a nutrient solution containing small molecule peptide, a mask containing the same and a manufacturing method. The mask is prepared from the following ingredients in parts by weight: the nutrient solution containing the small molecule peptide, water, hydroxyethyl cellulose, xanthan gum, glycerol, 1,3-butanediol, 1,2-hexanediol, p-hydroxyacetophenone, sodium hyaluronate, erythritol, sclerotium gum, EDTA disodium, arginine, arbutin, ethyl ascorbic acid, tremella extract, radix gentianae extract, camellia flower extract, hydroxylethyl urea, hydrogenated castor oil and pelargonium graveolens flower oil. The nutrient solution contains the small molecule peptide with extremely high biological activity; as a molecular weight of the small molecule peptide is small, the small molecule peptide can directly enter cells in original shapes through membranal permeation, further participates in enzyme synthesis, stimulates enzyme activity and enhances enzyme function; meanwhile, the small molecule peptide can further effectively provide nutrients for the cells and replace and eliminate toxin in the cells. The mask has varieties of effects of removing wrinkle, whitening skin, resisting ageing and the like.

The invention provides novel flavone hydroxylase, a microorganism for synthesizing a flavone C-glycoside compound and an application of the novel flavone hydroxylase. The inventor clones to obtain novel flavone hydroxylase PhF2H and PhF3'H, and the novel flavone hydroxylase PhF2H and PhF3'H belongs to cytochrome P450 hydroxylase and has a function of hydroxylating a specific position of the compound. Furthermore, the inventor efficiently synthesizes the flavone C-glycoside compound, such as orientin, isoorientin, vitexin and isovitexin, and related intermediates such as eriodictyol, 2-hydroxynaringenin and the like in a synthetic route of the flavone C-glycoside compound in an artificial recombinant expression system by modifying the enzyme and combining with the assembly of a synthetic route of C-glycoside glycosyltransferase and a flavone precursor.

The invention relates to Issatchenkia orientalis and a method for producing citicoline by whole cell conversion of Issatchenkia orientalis, belonging to the technical field of biological pharmacy. In the method provided by the invention, the whole cells of Issatchenkia orientalis Z1, namely CCTCC (China Center for Type Culture Collection) NO: M2011272 are utilized to prepare citicoline, choline phosphate and 5'-cytidylic acid are used as substrates, glucose is used as an energy donor, and the ATP (adenosine triphosphate) regeneration efficiency is improved by adding inorganic ions; glucose is used as the energy donor, so that the energy requirement of the strain is provided and ATP is provided for a citicoline synthesis enzyme system; one or more of potassium ion, magnesium ion and manganese ion are added to change the metabolism flow direction and improve the ATP regeneration rate, so that the ATP regeneration rate is matched with the rate of the citicoline enzyme synthesis system and the high-efficiency preparation of citicoline is achieved; and the whole cells of Issatchenkia orientalis are used, and toluene is added to an aqueous solution during the preparation process so as to improve the cell permeability, so that the rate of the citicoline enzyme synthesis system is improved.

The invention discloses a fungus with straw degradation capability and application thereof in composting and relates to the technical field of biological environmental protection. The straw degradation fungus JSD-17 is penicillium oxalicum (Penicillium oxalicum) and has a preservation number of CGMCC No.7743; the straw degradation fungus is further used for fermenting and composting agricultural waste straw. The penicillium oxalicum disclosed by the invention not only has strong cellulose enzyme synthesis capability, but also has the capability of decomposing hemicellulose and pectin components in the straw. By adopting the fungus disclosed by the invention, laccase (Lac), lignin peroxidase (Lip) and manganese peroxidase (Mnp) can be synthesized; the degradation speed of lignin components in the straw can be greatly increased.

An analytical system for rapid detection and identification of different analytes directly from a test sample by mixing test material with a germinogenic source and enzyme-free spores, allowing the mixture to stand for a time to allow analyte-induced spore germination and subsequent de novo activity of an enzyme capable of producing a germinant in the presence of the germinogenic source and detecting the presence of a germination-derived product. The germinant which is formed promotes further spore germination with concomitant additional de novo enzyme synthesis or activation which results in a propagating cascade of analyte-independent germination after which a germination-derived product can be easily detected. The technique is particularly efficient to conduct thousands of parallel assays in an array of microscopic wells.

The invention discloses a synthesis method of a disialic acid tetrasaccharide MAG antagonist by a chemical enzyme process. The invention also discloses two intermediates for synthesizing and preparing a tetrasaccharide MAG antagonist, which are respectively compounds disclosed as general formulae IV and V. By combining the flexibility of the chemical synthesis process and the regioselectivity and high efficiency of the enzyme synthesis process, the method overcomes the defects of low substrate reaction activity, multiple synthesis steps and low yield in the chemically synthesized disialic acid tetrasaccharide MAG antagonist at present, and the defects of difficulty in obtaining sialyl transferase, only recognition of glycopeptide and the like in the enzyme process, and has the advantages of high substrate reaction activity and high yield. Therefore, the method has important meanings in researching interactions between the sialic acid antagonist and the myelin related glycoprotein on the molecular level.

The invention discloses a preparation method of eco-friendly efficient compound foliar fertilizer. According to the characteristics of crop growth and development, various nutrients such as conditioning substances and nutrients are used according to a ratio so that the compound foliar fertilizer is formed. The compound foliar fertilizer contains complete nutrients and has multiple functions and strong specificity. Through use of a certain amount of a chelating agent, a surfactant or carrier, after spraying, the foliar fertilizer can be well adhered to and spread on the leaf surface so that leaf absorption and use are promoted. The rare earth elements lanthanum and cerium can effectively promote seed germination, promote crop stem growth, increase dry weight of seedlings, promote root growth, increase root activity and chlorophyll content, promote enzyme synthesis and pollen tube elongation, improve a crop seed setting rate, improve crop photosynthesis efficiency and a crop yield and improve the quality of crops. Through synergism of trace element selenium and gibberellin, a fungus inhibition capacity and crop resistance are improved, synthesis of chlorophyll is adjusted and crop quality is improved.

The invention relates to the phycocyanin fluorescent protein field, and particularly relates to a method for production of a streptavidin labeled phycocyanin fluorescent protein and an application. A streptavidin synthetic gene, a phycocyanin subunit synthetic gene, a catalytic enzyme synthetic gene and a chromophore synthetic gene are recombined in a same expression plasmid, and combination expression of multiple genes is realized; through optimization of induction and purification conditions, the fusion protein having stable fluorescent activity is prepared. Combination expression of fluorescent activity fusion phycobiliproteins are completed by adopting the single plasmid, the problems that the stability of multiple plasmids in an engineering strain and non-synchronization of expression of genes are avoided, and the use of antibiotics in the fermentation process is reduced. The method can effectively improve the stability of the fluorescent protein, besides, greatly reduces the production cost of phycocyanin fluorescent labeled preparations, and has good practical value and application prospects in the biomedical field, particularly in the production of fluorescent probes.

The invention discloses a synthesis method and an application of a sialylated TF antigen and its fluorination derivatives. The method includes the following steps: (1) chemically synthesizing fluorogalactose and fluorogalactosamine analogues; (2) chemically synthesizing a fluorinated TF antigen; and (3) synthesizing the sialylated TF antigen and its fluorination derivatives through an enzyme technology. The flexibility of a chemical synthesis technology is combined with the high regioselectivity and the high efficiency of the enzyme synthesis technology, so the enzymatic synthesis of the fluorosialylated TF antigen is achieved for the first time, and the disadvantages of many synthesis steps, poor stereoselectivity, low yield and use of a heavy metal salt in existing chemical synthesis ofthe fluorosialylated TF antigen are overcome. A fluorotumor-associated carbohydrate antigen has a higher stability than natural carbohydrate antigen, so the sialylated TF antigen and its fluorinationderivatives have a broad application prospect in the development of novel antitumor vaccines.

This invention relates to a recombinant microorganism capable of producing isoprene and isoprene production with the use of such recombinant microorganism with good efficiency. In this invention, the acetoacetyl-CoA synthase gene encoding an enzyme capable of synthesizing acetoacetyl-CoA from malonyl-CoA and acetyl-CoA and one or more genes involved in isoprene biosynthesis that enables synthesis of isoprene from acetoacetyl-CoA are introduced into a host microorganism.

The invention relates to a method for taking paper pulp as raw materials and as a carbon source which is needed for synthesizing cellulose enzyme by trichoderma reesei after the paper pulp is processed with dilute sulfuric acid. Compared with a method for taking pure cellulose as the carbon source, the filter paper enzyme activity can be increased by 40% through using the method for preparing the cellulose enzyme, and the problems that somatic cell is lack of the ability to utilize cellulose and the activity of the cellulose enzyme is not high when the paper pulp and dextrose are taken as the carbon source to prepare culture medium are solved.

The invention discloses a method for recovering D-p-hydroxyphenylglycine (D-HPG) from a cefprozil production waste liquid in an enzyme synthesis process, which comprises the following two steps: 1. separating D-HPG from the crystallization waste liquid through ion exchange by using an ion exchange resin; and 2. purifying and eluting the D-HPG by using hydrochloric acid solutions with different concentrations under the actions of ion exchange and static electricity, and drying to obtain the finished product. The method is characterized in that the process of separating and purifying the D-HPG from the cefprozil production waste liquid in the enzyme synthesis process is completed by using the ion exchange resin, and meanwhile, the subsequent waste liquid treatment technique of the enzyme-process cefprozil synthesis is completed.

Hydroxycarboxylic acids are produced by using a microorganism that is improved in ability to produce nicotinamide adenine dinucleotide by deleting, mutating or substituting nadR gene in the microorganism or introducing a gene encoding nicotinic acid phosphoribosyltransferase.

The invention relates to the field of recycling of cefalexin, in particular to a method for recycling cefalexin from an enzyme synthesis type cefalexin mother liquor. The method comprises the following steps of (1) separating the acidified cefalexin mother liquor by a film, so as to obtain a concentrated solution of the cefalexin; (2) enabling the obtained concentrated solution of the cefalexin instep (1) to be in contact with an alkaline regulator, and crystallizing, so as to obtain a crystal slurry of the cefalexin. The cefalexin obtained by the method has the characteristics that the purity is high, and the recycling rate is high; the preparation technology is simple, the cost is lower, and more environment-friendly effect is realized; under more preferable conditions, such as the stirring condition, the crystal is separated out, and the stirring phase is set to the phase 2 or phase 3, so that the granularity of the obtained cefalexin crystal is more uniform.

The invention discloses an engineering bacterium, and an application thereof in the production of Danshensu by using a cheap substrate, and belongs to the technical field of bioengineering. The engineering bacterium provided by the invention is a recombinant bacterium capable of producing Danshensu at a low cost; the recombinant bacterium can simultaneously express four enzymes which are tyrosinephenol lyase, L-amino acid oxidase, L-lactate dehydrogenase and alpha-hydroxycarboxylic acid oxidase respectively; and the recombinant bacterium knocks out a phenolic substance-decomposing gene, and can achieve enhanced expression of any one or more of a lactic acid transporter gene, a catechol transporter gene and a coenzyme synthesis-related gene. The engineering bacterium has the advantages ofrealization of the efficient production of Danshensu, simple process, few impurities and great industrial application values.