The way to obtain 6-oxycodonebuy acid
(57) Abstract:Usage: microbiological industry. The inventive method of obtaining 6-oxycodonebuy acid of nicotinic acid by enzymatic hydroxylation in the presence of microorganisms of the genera Pseudomonas or Achromobacter, in particular Pscedomonas acidovorans DS M 4746 or Achromobacter xylosoxydans DS M 2783. By complying with certain limits of concentration (from 0 to 10 g/l) for adding nicotinic acid is able to carry out the multiplication of the microorganism and to obtain the target product at one stage of the process without loss of product. The invention relates to a method for producing 6-oxycodonebuy acid enzymatic hydroxylation of nicotinic acid.Known methods for producing 6-oxycodonebuy acid using living microorganisms of the genera Pseudomonas, Bacillus or Achromobacter (patent Switzerland N 658866). According to the method of using a suspension of biomass relevant microorganisms that receive separately by multiplication of the starting culture. The hydroxylation process carried out periodically by a single addition of nicotinic acid as sodium salt. Due to the fact that in this way because p is enact a separate preliminary stage to obtain the total number of effective biomass.The purpose of the invention is the development of a more simple way, which receive a high concentration and output 6-oxycodonebuy acid.Discovered that it is possible to dispense with separate biomass production, if on the basis of obtained in a known manner starter cultures nicotinic acid be added so that its concentration in the suspension of microorganisms was mainly supported below the concentration above which retards the growth of microorganisms. The expression "mainly" means that the concentration of short-term and/or topically, in particular, in place of adding acid to its mixing with the suspension may exceed the maximum permissible value. In these conditions it was possible to expect formation of biomass, but suddenly it turned out that during this phase of growth is produced 6-oxycotine acid, which does not undergo further metabolisation, so after adding nicotinic acid and after complete consumption of 6-oxycodoneao acid it is possible to select virtually quantitative yield. Increasing with the addition of nicotinic acid concentration of 6-oxycodonebuy acid inhibits the growth of cells. However, the latter olisaemeka strains of microorganisms, these concentrations are about 50 g/l or more. Suddenly, what then 6-oxycotine acid is no longer subject to decay, but the inhibition of hydroxylase, i.e., enzyme, hydroxyperoxide nicotinic acid, and therefore, the limit of attainable concentration of the target product at 100% output only at higher concentrations. Significantly for the proposed method to the concentration of nicotinic acid for growth of the cells was never reduced to zero, because otherwise the decomposition of the formed 6-oxycodonebuy acid. Only when the concentration of 6-oxycodonebuy acid reaches the limit value, inhibited its degradation, so that after the addition of nicotinic acid can wait for its full flow without losses in output. As microorganisms, hydroxyperoxy nicotinic acid, it is advisable to use such genera as Pseudomonas, Bacillus, Achromobacter. Preference is given to the species Pseudomonas pituda and Achromobacter xylosoxydans, in particular the strain Achromobacter xylosoxydans DSM2783. Preferred microorganisms are described in the patent Switzerland N 658866. The concentration of nicotinic acid in the suspension of microorganisms for the entire period of its addition is less than 10 g/is ptx2">Nicotinic acid can be introduced in solid or dissolved form, in each case in the form of the free acid or water soluble salt. Preference is given to adding the aqueous solution, in particular an aqueous solution of sodium or potassium salt.It is advisable to aerate and stir the suspension of microorganisms, but can be limited to a mixing action of the blown air. The partial pressure of dissolved oxygen (po2should range 1-200 mbar, preferably 40 to 80 mbar. Preferably using a mechanical stirrer. The process of reproduction of microorganisms and getting 6-oxycodonebuy acid it is advisable to carry out at 20-40aboutC and pH 5.5-9.Together with nicotinic acid to promote the growth of biomass, it is recommended to add other nutrients. These include carbon sources such as glycerol or glucose, nitrogen sources such as ammonium salts or glutamic acid, and mineral salts, trace elements and vitamins. These substances should be added in such quantities that their concentration in the suspensions of the microorganisms were neither limiting nor in the acts of the process pretalk such or in the form of complex natural or obtained by synthetic mixtures. Particularly preferred complex nutrient is a yeast extract.The proposed method can be carried out continuously so that upon reaching the concentration of 6-oxycodonebuy acid, high enough to prevent its further decomposition to other products, begin to separate the target product with simultaneous recirculation cells. This can be accomplished, for example, continuous or periodic centrifugation or ultrafiltration.In order to maintain a constant concentration of the target product feed rate nicotinic acid should be equal to the speed of separation of the product.P R I m e R 1. a). Obtain starter cultures
From 5,19 g dihydrate sodium hydrogen phosphate, 2.0 g of potassium dihydrophosphate, 0.25 g yeast extract, and 1.00 g of nicotinic acid and 500 ml of water prepared liquid nutrient medium, which for 20 min sterilized at 120aboutC. After cooling medium 30aboutSince it was introduced a sterile concentrated solution of trace elements in number, ensuring the achievement in the medium specified concentration, mg/l: Dihydrate calcium chloride 20 Sulfate Mar is 11) 0.1 Heptahydrate zinc sulfate 0.1 molybdate Dihydrate sodium 0,1
Nutrient medium were seeded strain Achromobacter xylosoxydans DSM 2783. The culture was grown for 24 h at 30aboutC and pH 7.b). Received 6-oxycodonebuy acid.In the fermenter with a capacity of 20 l equipped with a stirrer, aeration system and device for regulating the pH value, in 12 l of water was dissolved in 90 g of nicotinic acid, 19,44 g of sodium hydroxide, 90 g of yeast extract, 12 g of potassium sulfate, 9.6 g of hexanitrate magnesium chloride, 1.92 g of calcium chloride, 2.4 ml of polypropylenglycol 2000, 180 g of L-glutamic acid and 300 g of glucose.The solution was sterilized for 30 min at 121aboutC. After cooling the solution to 30aboutSince it was introduced starter culture and with the air supply and mixing were cultured for 10 h at pH 7. After this period of time the concentration of nicotinic acid decreased from 7.5 to 2 g/l (according to a high-speed liquid chromatography), and the biomass concentration increased to 10 g/l (dry matter). At this point, began to add sterilized for 20 min at 121aboutTo a solution of 1.13 g of nicotinic acid and 0,365 kg of sodium hydroxide in 3 liters of water. The rate of addition was set so that the concentration of nicotinic acid in the farm, the new acid and glucose had been completely consumed and the concentration of biomass increased to 15 g/l (dry matter). After additional 4 h, i.e., through a total of 25 h after seeding, stopped fermentation. The final concentration of 6-oxycodonebuy acid was 74 g/l, which corresponds to a virtually quantitative conversion of nicotinic acid 6-oxycodonebuy. The reaction suspension was subjected to centrifugation to distinguish cells. Transparent centrifugal brought to a pH of 1.5 by addition of conc.hydrochloric acid, and 6-oxycotine acid was precipitated as a white solid. The product is filtered, washed with water and dried at 60aboutC and pressure of 20 mbar.P R I m m e R 2. a). Obtain starter cultures.Analogously to example 1A) made of a liquid nutrient medium, were seeded strain of Pseudomonas acidovorans DSM 4746. The culture was grown for 24 h at 30aboutC and pH 7.b). Getting 6-oxycodonebuy acid.The fermenter with a capacity of 20 l described in example 1 b), were loaded with liquid nutrient medium and sterilized as described in this example. The composition of the liquid nutrient medium was identical to example 1B) except that did not contain glucose.After cooling the solution to 30aboutSince it was introduced starter culture and with the air supply and peremeschennoe acid decreased from 7.5 to 2.5 g/l (according to a high-speed liquid chromatography), as the biomass concentration increased to 9.5 g/l (dry matter). At this point he started to continuously add a solution of nitronicotinic [receive and composition are consistent with those in example 1B)]. The rate of addition was set so that the concentration of nicotinic acid in the fermenter during the adding of the solution was between 1 and 9 g/l After 12 h the process of adding was completed, glutamic acid was completely consumed and the concentration of biomass increased to 14 g/l (dry matter).After additional 4 h, i.e. over the entire 24 h after the veil was stopped fermentation. The final concentration of 6-oxycodonebuy acid was 73 g/l, which corresponds to a virtually quantitative conversion of nicotinic acid 6-oxycodonebuy. Processing of the product was carried out as in example 1. The WAY to OBTAIN 6-OXYCODONEBUY ACID by microbiological hydroxylation of nicotinic acid by a microorganism of the genus Pseudomonas or Achromobacter under aerobic conditions at 20 - 40oC and at pH values of 5.5 to 9, characterized in that the microorganism used one of the strains of Pseudomonas acidovorans DSM 4746 or Achromobacter xylosoxydans DSM 2783, nicotinic acid or its soluble salt or its solution to what you are culture in the process of adding support in the interval 0 - 10 g/l, at least until the braking formed 6-oxycodonebuy acid on the growth of the microorganism and the process is conducted with simultaneous multiplication of the microorganism and education 6-oxycodonebuy acid.
SUBSTANCE: 4-[4-[4-(hydroxybiphenylmethyl)-1-piperidinyl]-1-hydroxybutyl]-α,α-dimethylphenylacetic acid is produced by incubation of mixture, containing terphenadine and microorganism capable to produce said acid followed by isolation of target product.
EFFECT: simplified process with increased yield.
20 cl, 2 tbl
SUBSTANCE: invention relates to isolated nucleic acid sequence encoding of polypeptide with nitrilase activity, wherein nitriles are converted to carboxylic acids in presence of said nitrilase.
EFFECT: method for production of chiral carboxylic acids with high effectiveness and low cost.
10 cl, 4 dwg, 2 tbl, 1 ex
FIELD: chemistry; biochemistry.
SUBSTANCE: invention relates to biotechnology, particularly to a method of obtaining ethylene unsaturated amide or ethylene unsaturated carboxylic acid or its salt from the corresponding ethylene unsaturated nitrile, in which the nitrile is hydrated or hydrolysed in an aqueous medium in the presence of a biocatalyst, where the nitrile contains more than 2 parts by weight per million acrolein and amide or carboxylic acid or its salt contains 2 parts by weight per million acrolein. The method can be used to obtain acrylamide or acrylic acid (salt) of high purity from low-quality acrylonitrile containing a lot of acrolein. Suitable biocatalysts include Rhodococcus bacteria.
EFFECT: invention increases efficiency of producing ethylene unsaturated amide or ethylene unsaturated carboxylic acid or its salt from the corresponding ethylene unsaturated nitrile and provides a method of producing a polymer of ethylene unsaturated monomer.
26 cl, 4 ex
SUBSTANCE: disclosed composition contains the following mixture, wt %: potassium succinate 40-80, potassium formate 10-30, potassium acetate 10-30, of the weight of the mixture with respect to dry substance. The disclosed aqueous composition contains the following mixture, wt %: potassium succinate 60-80, potassium formate 10-20, potassium acetate 10-20, with concentration of the mixture in water of 30-60%. Overall percentage content of potassium acetate and potassium formate in said compositions is essentially the same. Preferably, the mixture is obtained at least partially from a fermentation culture medium containing a carbon source and carboxylic acid-producing microorganisms.
EFFECT: composition has effective anti-icing and heat-exchange properties along with resistance to flaking of the concrete of an air-strip and corrosion inhibition.
23 cl, 10 dwg, 3 tbl, 17 ex
SUBSTANCE: invention describes a lipolytic polypeptide and a method for production thereof, involving: 1) collecting parent lipase with an amino acid sequence which is at least 80% identical to the sequence SEQ ID NO:1, which is given the description; 2) collecting at least one amino acid residue in the sequence; 3) altering the amino acid sequence, where the alteration includes one or more of the following: N74Q, P143S, A281S, P38S, N292Q, L1QGPL, L1QL, I285E, L147F, L147N, N292C, L140E, P143L, A146T, P280V, A283K, A284N, T103G + A148P, W104H + A148P, N74Q + A281S, V190A, L199P, T256K, T42N, R242A, V2151, T164V, L163F + T164V, D265P, P303K, R168D, A25G, V315I, T244P, K13Q, L277I, Y91S + A92S + N96S + N97* + L99V or D223G; 4) obtaining the altered polypeptide with the altered amino acid sequence; 5) determining specific activity of the lipolytic enzyme; and 6) collecting the altered polypeptide, having higher specific activity of the lipolytic enzyme than the parent polypeptide. The invention also discloses methods of conducting reactions which are catalysed by the described lipase, which involve reaction of reactants with said polypeptide, where the reaction is: hydrolysis of a carboxylic ester, synthesis of a carboxylic ester, alcoholysis of a carboxylic ester or acidolysis of a carboxylic ester.
EFFECT: invention enables to obtain lipase with higher specific activity than the parent lipase.
9 cl, 5 tbl, 1 dwg, 8 ex
SUBSTANCE: group of inventions relates to succinic acid-producing mutant microorganism, which is able to apply simultaneously saccharose and glycerine as carbon sources. Mutant microorganism is obtained by weakening of mechanism of saccharose-mediated catabolite repression of glycerol by removal of gene, which codes fructosephosphotransferase, or by introduction of gene, which codes glycerolkinase. Mutant microorganism is selected from the group, which consists of Mannheimia sp., Actinobacillus sp. and Anaerobiospirillum sp. Also claimed is method of obtaining mutant microorganism and versions of method of obtaining succinic acid with application of claimed microorganism.
EFFECT: group of inventions provides obtaining succinic acid with high output 1,54 mol/mol.
11 cl, 4 dwg, 2 tbl, 5 ex
SUBSTANCE: what is presented is a method for producing organic compounds. The method involves biomass fermentation in a fermenting box with volatile organic compounds generated, removal of the volatile organic compounds by gas stripping by means of a carrier gas, adsorption of the volatile organic compounds from a gas flow, desorption of the volatile organic compounds from the adsorbent, a catalytic reaction of the volatile organic compounds. A catalyst is zeolite, whereas the volatile organic compounds are alcohols, and/or ketones, and/or aldehydes, and/or organic acids.
EFFECT: high yield of the organic compounds and low equipment investments.
14 cl, 10 dwg, 1 tbl, 5 ex
SUBSTANCE: invention relates to biotechnology, particularly, to strains of fungi-producers of biologically active substances. Strain of Laetiporus sulphureus 3 having wide range of different biologically active substances, is deposited in Russian national collection of Industrial Microorganisms under registration number VKPM F-1188 and can be used in production of biologically active additives.
EFFECT: invention increases output of true protein and content of carotenoids.
1 cl, 3 tbl, 1 ex,
SUBSTANCE: invention relates to biotechnology. Disclosed is a method of converting lignocellulose in an organic acid. Method comprises pretreating lignocellulose material with an alkaline agent comprising containing calcium or magnesium cations in presence of water at a temperature of pretreatment from 20 to 115 °C. Aqueous suspension of pretreated material is obtained. At least part of suspension is fed into a fermentation zone. Method includes enzymatic hydrolysis and fermentation to produce broth containing insoluble lignocellulose, precipitated and dissolved calcium or magnesium salt of an organic acid and an enzyme. Enzymatic broth is unloaded, followed by separation of a liquid phase containing dissolved salt of an organic acid, and a solid phase containing insoluble lignocellulose and a precipitated salt of an organic acid. At least part of liquid phase is recycled to pretreatment step with alkali, and/or to fermentation zone.
EFFECT: invention enables to obtain a product in highly concentrated form with high output and purity.
16 cl, 1 dwg
SUBSTANCE: ascomycetous fungi strain of Sordariomycetes INA 01108 class has the ability to produce eremoxylarin A antibiotic. Ascomycetous fungi strain of Sordariomycetes class is deposited in the Russian National Collection of Microorganisms of the Federal State Budget Institution Science Institute of Biochemistry and Physiology of Microorganisms named after G.K. Skryabin, RAS under accession number VKM F-4676D and can be used in medicine.
EFFECT: invention allows to increase the yield of eremoxylarin A antibiotic.
4 dwg, 2 tbl, 1 ex
SUBSTANCE: group of inventions refers to getting 2.4-dihydroxybutyric acid (2.4-DHB). A method of obtaining 2.4-DHB, which includes the first stage of the transformation of malate in 4-phosphomalate using an enzyme, capable to carry out such conversion, the second stage of the transformation of 4-phosphomalate in malate-4-half-aldehyde by using an enzyme capable to implement a similar metamorphosis, and third stage of transformation of malate-4-half-aldehyde in 2.4-DHB using an enzyme, capable to carry out such conversion. At the first stage, using an enzyme that has a sequence selected from the group consisting of SEQ ID No. 12, SEQ ID No. 14, SEQ ID No. 16, SEQ ID No. 18, SEQ ID No. 20, SEQ ID No. 22, SEQ ID No. 24, SEQ ID No. 26, SEQ ID No. 39, SEQ ID No. 41, SEQ ID No. 43 and SEQ ID No. 45. The second stage uses an enzyme that has a sequence selected from the group consisting of SEQ ID No. 54, SEQ ID No. 56, SEQ ID No. 58, SEQ ID No. 60, SEQ ID No. 62, SEQ ID no. 64, SEQ ID No. 66 and SEQ ID No. 231. In the third stage, using an enzyme that has a sequence selected from the group consisting of SEQ ID No. 74, SEQ ID No. 76, SEQ ID No. 81, SEQ ID No. 225 and SEQ ID No. 227. Proposed options for the enzymes used in the method, nucleic acid variants which encode the relevant enzymes variants chimeric genes for transformation of the micro-organism-host and microorganism in the expression of the corresponding enzyme expression vector variants, variants of a micro-organism-host expressing the relevant enzyme.
EFFECT: improved method.
31 cl, 5 dwg, 16 tbl, 12 ex