463 results about "Enzymatic synthesis" patented technology

The invention provides methods, kits, and compositions for enhancing synthesis of DNA involving a carboxylate ion-supplying substance that is effective in promoting DNA synthesis in enzymatic DNA synthesis reactions. The invention further provides a thermostable DNA polymerase-related factor derived from Thermococcus species, which has an activity to promote the DNA synthesis activity of DNA polymerase or which binds to DNA polymerase.

The present invention comprises methods and compositions for splint-assisted enzymatic synthesis of polyribonucleotides using an RNA polymerizing enzyme. The invention provides ligating ribonucleotides comprising ligating a donor RNA molecule to an acceptor RNA molecule in the presence of RNA ligase and a splint, wherein the donor RNA molecule is comprised of at least one nucleotide and a ligation linker moiety, the acceptor RNA molecule is comprised of at least one nucleotide and a ligation linker moiety and the splint is comprised of a polyribonucleotide. The invention also provides splints for use in splint-assisted enzymatic synthesis using an RNA polymerizing enzyme.

Methods for the synthesis of CCI-779 and proline-CCI-779 are described, including a method involving lipase-catalyzed acetylation of 42-hydroxy of rapamycin with a vinyl ester of 2,2-bis(hydroxymethyl) propionic acid in an organic solvent followed by deprotection. Also provided are products containing proline-CCI-779 and uses thereof.

The present invention relates to the enzymatic synthesis of oligosaccharides, including sialylated product saccharides. In particular, it relates to the use of recombinant cells to take up low cost precursors such as glucose, pyruvate and N-actyl-glucosamine, and to synthesize activated sugar moieties that are used in oligosaccharide synthesis. The methods make possible the synthesis of many oligosaccharides using microorganisms and readily available, relatively inexpensive starting materials.

This invention provides a direct method for monitoring bacterial transglycosylase activity using labeled substrates produced by chemo-enzymatic synthesis wherein the labels are selected to permit the detection of both polymeric and non-polymeric products simultaneously, either directly or following the separation of product from starting material. The invention promotes the discovery of new antibiotics with activity against bacterial transglycosylases by a) laying the groundwork for structural analysis of purified, active transglycosylase (which permits structure-based design); and b) providing an assay that can be used to screen for inhibitors.

A novel UDP-Gal regeneration process and its combined use with a galactosyltransferase to add galactose to a suitable acceptor substrate. Also described herein are synthetic methods for generating Globo-series oligosaccharides in large scale, wherein the methods may involve the combination of a glycosyltransferase reaction and a nucleotide sugar regeneration process.

In one aspect the present invention provides methods of synthesizing a preparation of nucleic acid molecules, the methods comprising the steps of: (a) utilizing an RNA template to enzymatically synthesize a first DNA molecule that is complementary to at least 50 contiguous bases of the RNA template; (b) utilizing the first DNA molecule as a template to enzymatically synthesize a second DNA molecule, thereby forming a double-stranded DNA molecule wherein the first DNA molecule is hybridized to the second DNA molecule; (c) utilizing the first or second DNA molecule of the double-stranded DNA molecule as a template to enzymatically synthesize a first RNA molecule that is complementary to either the first DNA molecule or to the second DNA molecule; and (d) utilizing the first RNA molecule as a template to enzymatically synthesize a third DNA molecule that is complementary to the first RNA molecule. In another aspect, the present invention provides processed DNA samples prepared by a method of the invention for synthesizing a preparation of nucleic acid molecules. In another aspect, the present invention provides methods for hybridizing a processed DNA sample to a population of immobilized nucleic acid molecules.

Poly(amine-co-ester) polymers, methods of forming active agent-load nanoparticles therefrom, and methods of using the nanoparticles for drug delivery are disclosed. The nanoparticles can be coated with an agent that reduces surface charge, an agent that increases cell-specific targeting, or a combination thereof. Typically, the loaded nanoparticles are less toxic, more efficient at drug delivery, or a combination thereof compared to a control other transfection reagents. In some embodiments, the nanoparticles are suitable for in vivo delivery, and can be administered systemically to a subject to treat a disease or condition.

The present invention provides methods for producing sialyloligosaccharides in situ in dairy sources and cheese processing waste streams, prior to, during, or after processing of the dairy source during the cheese manufacturing process. The methods of the present invention use the catalytic activity of alpha(2-3) trans-sialidases to exploit the high concentrations of lactose and alpha(2-3) sialosides which naturally occur in dairy sources and cheese processing waste streams to drive the enzymatic synthesis of alpha(2-3) sialyllactose. alpha(2-3) sialyloligosaccharides produced according to these methods are additionally encompassed by the present invention. The invention also provides for recovery of the sialyloligosaccharides produced by these methods. The invention further provides a method for producing alpha(2-3) sialyllactose. The invention additionally provides a method of enriching for alpha(2-3) sialyllactose in milk using transgenic mammals that express an alpha(2-3) trans-sialidase transgene. The invention also provides for recovery of the sialyllactose contained in the milk produced by this transgenic mammal either before or after processing of the milk. Transgenic mammals containing an alpha(2-3) trans-sialidase encoding sequence operably linked to a regulatory sequence of a gene expressed in mammary tissue are also provided by the invention.

Hydrazino, oxyamino and carbonyl-based monomers and methods for incorporation into oligonucleotides during enzymatic synthesis are provided. Modified oligonucleotides are provided that incorporate the monomers provided herein. Immobilized oligonucleotides and oligonucleotide conjugates that contain covalent hydrazone or oxime linkages are provided. Methods for preparation of surface bound oligonucleotides are provided. Methods for the preparation of oligonucleotide conjugates are also provided.

The invention discloses a load-type ion liquid catalyst and a preparation method and an application thereof. The load-type ion liquid catalyst is characterized in that a carrier is loaded with pyridine acidic ionic liquid [XPy]HSO4, wherein X is ethyl, propyl, isopropyl, normal-butyl, 3-methylbutyl or 3-amyl; the carrier is gamma-Al2O3, SiO2, mesoporous molecular sieve SBA-15 and carbon fiber; themass ratio of the carrier to the pyridine acidic ionic liquid is 0.5-1.5:1. The invention also discloses the preparation method of the load-type ion liquid catalyst and the application thereof in enzymatic synthesis of diglycol. The invention has the beneficial effects that the invention provides a novel load-type ion liquid catalyst, has simple catalyst preparing method, high catalyst activity and long service life, is applied to enzymatic synthesis of diglycol, has moderate reaction condition, simple production technology, higher diglycol percent conversion and selectivity and is suitable for industrialization.

Poly(amine-co-ester) polymers, methods of forming active agent-load nanoparticles therefrom, and methods of using the nanoparticles for drug delivery are disclosed. The nanoparticles can be coated with an agent that reduces surface charge, an agent that increases cell-specific targeting, or a combination thereof. Typically, the loaded nanoparticles are less toxic, more efficient at drug delivery, or a combination thereof compared to a control other transfection reagents. In some embodiments, the nanoparticles are suitable for in vivo delivery, and can be administered systemically to a subject to treat a disease or condition.

The invention discloses a recovering method of effective components in amoxicillin enzymatic synthesis mother liquor by utilizing nanofiltration. The method comprises the following steps: (1) regulating pH of the mother liquor; (2) hydrolyzing amoxicillin in the mother liquor by utilizing penicillin acylase, thus only D-hydroxyphenylglycine and 6-amino-penicillanic acid (6-amino-penicillanic acid) exist in the other liquor; (3) preparing concentrated mother liquor by nanofiltration; and (4) separating and recovering the D-hydroxyphenylglycine and the 6-amino-penicillanic acid (6-amino-penicillanic acid) by isoelectric point crystallization. The method has the advantages that: the recycling rate of an inactive lateral chain can be improved regarding certain amoxicillin synthetase with highhydrolysis activity, so that the enzymatic synthesis of amoxicillin is more economical.

The invention relates to a method for continuous enzymatic synthesis of n-3PUFA glyceride, comprising the following steps: preparing fish oil n-3PUFA (EPA, DHA) and glycerol into reaction liquid according to a certain proportion; then pumping reaction liquid into an enzyme reaction column provided with immobilized lipase through a constant current pump for carrying out reaction for enzymatic synthesis of n-3PUFA; finally realizing continuous enzymatic synthetic production of n-3PUFA by controlling circulation flow velocity of the reaction liquid in the reactor and setting a molecular sieve dehydrater in the reaction system. The n-3PUFA glyceride product prepared by the continuous syhthesis method features an ester yield of 30-50%, a monoester content of 20-30%, a diester content of 50-70%and a trig content of 10-20%. With the method of the invention adopted, recycle of enzyme can be realized, cost can be greatly reduced, production cycle can be shortened, production efficiency can beimproved and the operation is easy, so that the method is of great practical value to industrialized batch production.

The invention discloses a synthesis process for preparing (S)-2-aminobutyramide by enzymatic method. The current preparation method has problems such as high cost, low yield, and large environmental hazards. The technical scheme adopted in the invention is: using 2-aminobutyronitrile as a starting material to prepare (S)-2-aminobutyramide in one step in a buffer solution system through enzyme catalysis. In the present invention, water is used as a solvent in the main process, without using a large amount of organic solvents harmful to the environment, the whole process cost is low, and it is beneficial to large-scale industrial preparation.

The invention relates to a medicine synthesis method, in particular to a process method for screening of immobilized cefprozil synthetase and enzymatic synthesis of cefprozil. In order to solve the problems that a conventional cefprozil enzymatic synthesis process has difficulties in screening and evaluating immobilized anzyme, tedious in production process step, poor in control point, long in reaction time, low in conversion ratio and the like, the invention provides an immobilized cefprozil synthetase and a novel process for cefprozil synthesis.

The invention relates to a preparation method of sterol ester and in particular relates to an enzymatic synthesis method of phytosterol ester / phytostanol ester by utilizing ultrasonic enhancement. The method is characterized by comprising the following steps: 1) preparing a heterogeneous enzyme reaction system: weighing the phytosterol ester / phytostanol ester and fatty acid at a molar ratio of 1:(1.2-2), adding a reaction solvent, and adding enzyme and a molecular sieve; 2) performing enzymatic esterification with ultrasonic assistance: immersing an ultrasonic probe in a reaction solution, opening an ultrasonic wave generator, constantly radiating with ultrasonic wave for 0.5 to 3 hours under mechanical stirring, and continuously reacting for 4 to 10 hours under mechanical stirring after the radiation is completed; and 3) removing the fatty acid with ultrasonic assisted saponification to obtain the product. According to the method, the enzymatic esterification reaction time can be remarkably shortened, the usage of enzyme can be reduced, and the saponification speed of excessive fatty acid can be accelerated, so that the efficiency of separation and purification can be improved, and the obtained phytosterol ester / phytostanol ester product has the characteristics of light color, high purity and safety without contamination.

The invention discloses a production method of rare ginsenoside Rh2 and belongs to the field of enzymatic synthesis of ginsenoside. The production method includes that protopanoxadiol is used as a substrate, glycosyltransferase YjiC or YojK screened from bacillus subtilis is used as a catalyst, and the glycosylation of the C3 hydroxyl of the protopanoxadiol is efficiently catalyzed through in-vitro enzyme reaction to generate the ginsenoside Rh2. The production method is cheap in raw materials, high in ginsenoside Rh2 yield, low in production cost and capable of achieving large-scale production of the precious and rare ginsenoside Rh2.

The invention provides methods for purifying carbohydrates, including oligosaccharides, nucleotide sugars, and related compounds, by use of ultrafiltration, nanofiltration and / or reverse osmosis. The carbohydrates are purified away from undesired contaminants such as compounds present in reaction mixtures following enzymatic synthesis or degradation of oligosaccharides.

The invention discloses two ginsenoside Rh2 aliphatic ester compounds. The compounds have the following structural formula described in the specification. In the formula, R is a C4-C22 fatty acid acyl. The invention also provides the chemical synthesis and enzymatic synthesis methods of the ginsenoside Rh2 aliphatic ester compounds, a pharmaceutical composition which takes the compounds as active ingredients and an application of the compounds and the pharmaceutical composition in preparation of an anticancer drug.

The present invention provides methods for producing cannabinoids. More specifically, the invention is directed to the bio-enzymatic synthesis of THC-v, CBV and CBN by contacting a compound according to Formula I with a cannabinoid synthase enzyme. Also described is a system for producing these pharmaceutically important cannabinoids and the use of such cannabinoids as therapeutic agents.

Process for enzymatically synthesizing carboxylic esters, characterized in that mixing and discharge of the water of reaction are effected by introducing a gas while achieving low effectiveness ratios EV.

Heparin is synthesized from a polysaccharide comprised of a 1-4 glycosidically linked alternating polymer of uronic acid and glucosamine residues, wherein the uronic acid is selected from iduronic and glucuronic acid, wherein the glucosamine is partially N-sulfated; by a series of selective reactions catalyzed by recombinant enzymes.

The invention discloses a method for producing L-ascorbyl palmitate through non-aqueous phase enzymatic synthesis. The method comprises the following steps of: (1) mixing L-ascorbic acid with palmitic acid in a non-aqueous medium, adding immobilized lipase for catalytic reaction for 20-50 hours; (2) filtering the reaction liquid obtained in the step (1) to remove the immobilized lipase and undissolved L-ascorbic acid and evaporating the obtained filtrate under reduced pressure to remove a solvent; and (3) dissolving the residue obtained in the step (2) with ethyl acetate, washing with deionized water, separating an ethyl acetate phase out, evaporating under reduced pressure to remove the solvent, dissolving the residue with toluene, recrystallizing, centrifuging, washing the obtained substance with normal hexane and drying the obtained undissolved substance in vacuum to obtain an L-ascorbyl palmitate product. The method relates to non-aqueous enzymatic synthesis, has the advantages of high catalytic efficiency, mild reaction condition, low pollution, simple process, readily-available raw materials, high yield and convenience for large-scale production, and is accordant with the practical situation of the food additive production industry in China.

The invention discloses a ethyl lactate synthesis enzyme catalysis method in solvent phase and belongs to ethyl lactate preparation technology using lactic acid and ethanol, characterizing in the following steps: using chloroform, tert-butanol or tetrahydrofuran as sovent, or adding in the said solvent of lactic acid and ethanol in ratio of (1:5)-(1:10) and lipase according to 10-30 g enzyme catalysis per mol lactic acid, mixing sufficiently, constant temperature water bath in 60-70 DEG C while lactic acid concentration is 0.3-0.6mol / L, concussion table revolution being controlled 200-250revolution / min, reacting 24-48 hours, obtaining crude products containing ethyl lactate. Advantage of the invention compared with existing ethyl lactate preparation method lies in: temperate reaction condition, low energy consumption, easy operation of separating catalyst and products, apparatus not easy corroding, no pollution, high product quality and yield.

This invention discloses an improved enzymatic process, under organic solvent free conditions, for the incorporation of fatty acids such as omega-3 fatty acids into phospholipids. The rate of transesterification is increased 4 times by adding a base to the reaction mixture, typically an amine. The invention also discloses novel phospholipid compositions as well as novel use of the phospholipid compositions as a food supplement, a fish feed, animal feed and human food. In addition to methods for enriching prey organisms used in aquaculture, methods of reducing arachidonic acid levels in mammalian plasma / red blood cells and methods for increasing DHA levels in the mammalian brain.

Dietary supplements for treating a neurotransmitter deficiency include one or more precursors of the deficient neurotransmitter and a cofactor for activating in vivo enzymatic synthesis of the deficient neurotransmitter. The dietary supplements can also include an appropriate neurotransmitter, such as an amino acid. When administered through the oral mucosa, increases in levels of the deficient neurotransmitters and relief from deficiency symptoms are obtained.