51 results about "Cell free extracts" patented technology

Methods are provided for the utilization of bacterial cell-free extracts in the synthesis of high yields of virus like particles with encapsidated cargo.

Enzymatically active hydrogenase is synthesized in a cell-free reaction. The hydrogenases are synthesized in a cell-free reaction comprising a cell extract derived from microbial strains expressing at least one hydrogenase accessory protein. In some embodiments, the extracts are produced under anerobic conditions.

The invention discloses a method for preparing (R)-2-hydroxy-4-phenyl ethyl butyrate by catalyzing recombinant carbonyl reductase, which belongs to the technical field of biological engineering. The method comprises the following steps of: cloning gene segments of carbonyl reductase (IolS) and glucose dehydrogenase (GDH) from bacillus subtilis CGMCC NO.1.1508, expressing an IolS gene and a GDH gene in series by adopting a dual-starter method to construct a recombinant plasmid pET24a-G-T7-I, and introducing the plasmid into escherichia coli BL21(DE3); and under the condition of not adding or adding a small amount of NADP+cofactors, performing biotransformation by taking a cell-free extract of the escherichia coli recombinant plasmid as a catalyst, 2-oxo-4-phenyl ethyl butyrate as a substrate and glucose as a substrate to obtain (R)-2-hydroxy-4-phenyl ethyl butyrate, wherein the enantiomeric excess value of the product is higher than 99.5 percent. In the method, IolS and GDH are co-expressed, so that efficient regeneration of an intra-cellular cofactor NADP(H) is realized, production cost is lowered, and a good industrial application prospect is achieved.

A method and composition for treating dermatological conditions and improving skin condition and helping hair to grow or thicken involves putting mesenchymal stem cells or other cells with regenerative properties into culture medium to induce secretion of beneficial factors such as growth factor, regulatory factors, enzymes, hormones, peptides and lymphokines into the culture medium to create conditioned medium and then extracting either a supernatant or a cell free extract of the conditioned medium to use itself or components thereof alone or with other skin or hair care reagents as a topical ointment or formula to apply to the skin or hair topically for therapeutic and cosmetic purposes.

The present invention relates to novel mixtures comprising, as active components, at least one isolated bacterial strain, which is a member of the genus Paenibacillus, or a cell-free extract thereof or at least one metabolite thereof, and at least one other biopesticide. The present invention also relates to compositions comprising at least one of such bacterial strains, whole culture broth or a cell-free extract or a fraction thereof or at least one metabolite thereof, and at least one other biopesticide. The present invention also relates to a method of controlling or suppressing plant pathogens or preventing plant pathogen infections by applying such composition. The present invention also relates to mixtures of fusaricidins which are pesticidal metabolites produced by the abovementioned strains, and other biopesticides.

The present invention relates to a composition comprising a biologically pure culture of a fungicidal Paenibacillus sp. strain comprising a variant fusaricidin synthetase lacking a functional adenylation domain in the third module. The present invention also provides a composition comprising a biologically pure culture of a fungicidal Paenibacillus sp. strain or a cell-free extract thereof comprising at least one Paeniserine and at least one Paeniprolixin. Also provided are isolated compounds and methods of treating a plant to control a plant disease with the disclosed compositions and compounds.

The invention relates to novel polyenes having formula (I), wherein: R1 represents alkyl C₁-C₃; and R2 represents a functional group selected from CH₃— or CONH₂— (methyl- or primary amide-). The aforementioned polyenes have a biocide action on organisms comprising cell membranes that contain ergosterol, e.g., fungi or parasites. Said compounds can be obtained using a method that consists in cultivating a producing micro-organism under conditions that enable the production thereof. In addition, the invention also relates to a mechanism for the in vitro production of amidated polyenes, consisting in incubating carboxylated polyenes with cell-free extracts (or proteinaceous fractions) of the producers of same in the presence of ATP/Mg⁺⁺ and an amide- group donor compound (preferably glutamine).

The invention relates to methods for performing in vitro site-directed mutagenesis of a targeted gene or genes. In another aspect, the invention includes in vitro site-directed mutagenesis kits comprising a ribonucleotide particle (RNP), an oligonucleotide, a buffer, a cell-free extract, and instructional material for use thereof.

Insect cells are stored in a small gas cylinder, and the small gas cylinder is charged with nitrogen gas to pressurize the cylinder. The charged gas is exhausted at once to crush the cells to provide the objective cell extract with translation activity and glycosylation activity. As this method is gentler than the conventional cell-crushing method employing a homogenizer, in addition to translation factors, factors carrying glycosylation activity can also be recovered. As a result, an in-vitro glycoprotein synthesis system capable of performing translation to post-translation glycosylation can be produced.

The present disclosure provides methods for incorporating at least one non-natural amino acid into a polypeptide using a cell-free protein synthesis which includes a cell-free extract and is deficient in endogenous tRNA. The methods include providing the synthesis system with at least one non-natural amino acid, at least one orthogonal tRNA and one orthogonal aminoacyl-tRNA synthetase which aminoacylates the corresponding orthogonal tRNA with the non-natural amino acid. The methods may also include removing the endogenous tRNA in the cell-free protein synthesis system by treating the synthesis system with a ribonuclease to degrade the endogenous tRNA and providing the synthesis system with a minimal set of tRNAs for one or more natural amino acids and the corresponding amino-acyl tRNA synthetases, such that the lack of some tRNA will enable unique codons to encode for non-natural amino acid incorporation with no or a minimal amount of completion from the tRNA present.

The invention relates to the technical field of microbiological screening and application and in particular provides a novel lactic acid bacterium preparation. The lactic acid bacterium preparation can be widely applied to treatment of gastric ulcer, the curative effect can be obviously improved, the cure rate of the ulcer is 90%, the eradication rate of helicobacter pylori is 84%, and the lactic acid bacterium preparation is high in tolerance to treatment and suitable for clinical application and popularization. The lactobacillus fermenti selected in the lactic acid bacterium preparation has obvious inhibitory effects on escherichia coli, salmonella and staphylococcus aureus, particularly has the strongest inhibitory effect on helicobacter pylori, and the diameter of an inhibition zone is 23mm. The lactobacillus fermenti has high tolerance to gastric acid and bile salt, and can realize effective multiplication in a gastric acid environment. The removal rate of lactobacillus fermenti KDB-8 on cholesterol is 93.2%; the survival rate of 98.5% can be maintained in 1.2mmol/L H2O2; and the scavenging rate of fermented supernatant fluid and cell-free extract on hydroxyl radicals is respectively 95.1% and 90.4%, and an undesirable technical effect is achieved.

The invention provides a method for compounding non-natural amino acids pPpa in escherichia coli aquaporin AQPZ. The method comprises the following steps: carrying out site-directed mutation on tyrosine-tRNA synthetase of methanococcus jannaschii to obtain an MjpPpaRS gene; cloning the MjpPpaRS gene onto a pIVEX2.4c plasmid to obtain a recombinant plasmid pIVEX2.4c-MjpPpaRS; converting the recombinant plasmid pIVEX2.4c-MjpPpaRS and pUC-MjtRNA to escherichia coli BL21; selecting a positive transformant and fermenting to prepare escherichia coli cell-free extract; establishing a cell-free expression system; taking a pIVEX2.4c-AqpZ as a template and carrying out amber mutation on three sites including F10, F13 and F17 of the AQPZ; adding a decontaminating agent into the cell-free expression system to dissolve and express the AQPZ programmed into the pPpa; and carrying out affinity chromatography purification to obtain the AQPZ which is programmed into the pPpa at a fixed site. The AQPZ programmed into the pPpa is produced through utilizing the escherichia coli cell-free expression system, and the target protein is obtained through utilizing the affinity chromatography purification; and the non-natural amino acids are embedded, so that the affinity of the aquaporin and phospholipid is enhanced and the aquaporin is more stable after being embedded under the same condition.

The invention relates to a method for biological catalyzed synthesis of optically active alkyl lactone. The method comprises the following steps: (1) catalyzing asymmetric reduction of a carbonyl acid to synthesize optically active hydroxy acid by using a biological catalyst; (2) in an acid environment, cyclizing the optically active hydroxy acid, thereby generating an optically active lactone compound. Freshly cultured candida parapsilosis cells or a cell-free extract is adopted as the biological catalyst for catalyzing stereoselective reduction of 4-carbonyl capric acid and 5-carbonyl capric acid, and subsequently the components are cyclized in the acid environment so as to obtain the lactone compound, so that (R)-(+)-4-hexyl-4-butyrolactone and (R)-(+)-5-amyl-5-valerolactone can be efficiently prepared. Compared with the prior art, the method is easy in preparing cells of the strain and extracts of the cells, high in stereoselectivity, simple in process for catalytic preparation of (R)-(+)-4-hexyl-4-butyrolactone and (R)-(+)-5-amyl-5-valerolactone, environment-friendly, and very good in industrial application prospect.

The present invention relates to novel isolated bacterial strains, which are members of the genus Paenibacillus, originally isolated from soil and showing antagonistic activity against a broad range of pathogens and being capable of producing antimicrobial metabolites. It was found that the strains Lu16774 and Lu17007 belong to a novel subspecies named Paenibacillus polymyxa ssp. plantarum while the strain Lu17015 belongs to a novel species which is proposed to be Paenibacillus epiphyticus. The present invention also relates to microbial pesticide compositions comprising at least one of such novel bacterial strains, whole culture broth or a cell-free extract or a fraction thereof or at least one metabolite thereof, and/or a mutant of at least one of said novel bacterial strains having all the identifying characteristics of the respective bacterial strain or whole culture broth, cell-free extract, fraction and/or metabolite of the mutant thereof showing antagonistic activity against plant pathogens. The present invention also relates to a method of controlling or suppressing plant pathogens or preventing plant pathogen infections by applying such composition. The present invention also relates to novel fusaricidin-type compounds which are metabolites produced by the strains of the present invention.

Methods are provided for the utilization of bacterial cell-free extracts in the synthesis of high yields of virus like particles.

The present invention refers to the strain CECT8836 of Bacillus amyloliquefaciens and mutants thereof, and the use of said strain as a pesticide in controlling plant diseases caused by fungi and bacteria. Further aspects of the invention relate to methods for preparing suspensions and extracts of the strain CECT8836 of B. amyloliquefaciens, pesticidal compositions comprising said strain and an extract of CECT8836 of B. amyloliquefaciens with antimicrobial activity. Finally the invention relates to a method for the biological control of various plant diseases caused by fungi and bacteria both in vegetable plants and in fruit trees, comprising treating these plants with the strain CECT8836 of B. amyloliquefaciens, a composition including it or a cell-free extract derived from CECT8836 of B. amyloliquefaciens.

The invention provides a method of inhibiting Ran protein activity in at least one eukaryotic cell or cell-free extract, the method comprising exposing an amino acid sequence comprising at least a portion of EMCV or TMEV leader protein, wherein the amino acid sequence comprises SEQ. ID NO: 14 or SEQ. ID NO: 15, to at least one cell in an amount effective to inhibit Ran activity in the targeted cell and evaluating Ran protein activity in the cell.