New fucosyltransferases and applications thereof

FIELD: biotechnology.

SUBSTANCE: bacterial host cell is provided comprising a sequence consisting of a polynucleotide encoding a polypeptide with alpha-1,2-fucosyltransferase activity (SEQ ID NO: 1) to produce 2'-fucosyllactose. Also methods for 2'-fucosyllactose production using the said polypeptide with alpha-1,2-fucosyltransferase activity, a donor substrate containing a fucose residue and an acceptor substrate comprising or consisting of lactose.

EFFECT: group of inventions allows to facilitate production of fucosylated oligosaccharides of human milk, making it more efficient and economical due to the possibility of lactose application as a substrate for alpha-1,2-fucosyltransferase.

9 cl, 13 dwg, 3 tbl, 1 ex

 



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. Disclosed is a method of producing β-cyclodextrin. The method includes preparing a suspension of maltodextrin, carrying out a ring closure process using a CGTase enzyme preparation in the presence of a trichloroethylene complexant. Further, the method includes removing the complexant by boiling, purifying the obtained product with activated carbon, separating the activated carbon by filtering and evaporating the syrup and crystallisation thereof while stirring and lowering the temperature. Finally, the method includes centrifuging and washing β-cyclodextrin crystals with cold water, drying and packaging.

EFFECT: invention can be used to produce cyclic complexing agents, particularly β-cyclodextrin.

2 dwg

FIELD: chemistry.

SUBSTANCE: disclosed is a method for in vivo large-scale synthesis of sialylated oligosaccharides and a microorganism containing heterologous genes which code GlcNAc-synthetase, sialic acid synthase, ClcNAc-6-phosphate-2-epimerase and sialyl transferase, wherein endogenous genes which code sialic acid aldolase (NanA) and ManNac-kinase (NanK) were delegated or inactivated.

EFFECT: invention can be used in industrial synthesis of sialyl galactose.

58 cl, 4 tbl, 20 dwg, 17 ex

FIELD: medicine.

SUBSTANCE: method involves biosynthesis of penta-N-acetylchitopentaose in vivo from N-acetylglucosamine precursor in the high-density bacterial culture Escherichia coli DH5α with the use of recombinant enzyme of N-acetylglucosaminyl transferase Mesorhizobium loti 1803 expressed in cells of said bacteria. The biosynthesis in vivo is enabled by recovery, cloning and expression in Escherichia coli DH5αa of the gene NodC Mesorhizobium loti 1803 coding N-acetylglucosaminyl transferase; the gene NodC is amplified with the use of specific primers and Pfu-polymerase; a matrix for amplification of the gene NodC is presented by DNA recovered from the strain Mesorhizobium loti 1803 deposited in the collection of All-Russian Research Institute for Agricultural Microbiology; it results in preparing an amplification product sized 1300 base pairs; the amplified gene NodC Mesorhizobium loti 1803 (NodC) is cloned in the vector pUC19 in sites BamH EcoRl with preserving a reading frame so that to produce the plasmid pUC19-NodC that is used for expression of the gene NodC in the bacteria Escherichia coli DH5α; N-acetylglucosaminyl transferase (NodC) is expressed in the strain Escherichia coli DH5a after induction by isopropyl-β-D-thiogalactoside in cultivation of the bacteria Escherichia coli DH5α bearing the plasmid pUC19-NodC with the built-in gene NodC Mesorhizobium loti 1803; neHTa-N-acetylpentachitopentaose is synthesised in the presence of the N-acetylglucosamine precursor; the amount of neHTa-N-acetylpentachitopentaose synthesised by the enzyme N-acetylglucosaminyl transferase Mesorhizobium loti 1803 makes about 0.1-0.5 gram per one litre of the bacterial culture; the synthesised substance is analysed by mass spectrometry to prove that the synthesised substance is penta-N-acetylchitopentaose.

EFFECT: technique improvement.

5 dwg, 2 ex

FIELD: biotechnology, food processing industry, biochemistry.

SUBSTANCE: invention elates to a method for enzyme hydrolysis of granular starch to soluble starch at temperature lower of the initial gelatinization point of indicated granular starch. Method involves simultaneous effect of the first enzyme representing a member of family 13 of glycoside hydrolase, it shows alpha-1,4-glucoside hydrolytic activity and comprises carbon-bound modulus of family 20 and at least one the second enzyme representing beta-amylase or glycoamylase. Invention provides carrying out hydrolysis of starch with the high degree of effectiveness.

EFFECT: improved method of treatment.

32 cl, 24 tbl, 9 ex

FIELD: genetic engineering, biochemistry.

SUBSTANCE: method involves transformation of plant with DNA encoding protein possessing with enzymatic activity of amylosaccharase. Interaction of protein obtained from plant with sucrose-containing solution provides conversion of sucrose to α-1,4-glucanes and/or fructose. Invention can be used in preparing linear α-1,4-glucanes in plants.

EFFECT: improved preparing method.

6 cl, 2 dwg, 6 ex

FIELD: biotechnology, biochemistry, microbiology.

SUBSTANCE: the strain Bacillus circulans B-65 a producer of cyclodextrin glucanotransferase is isolated and selected form the soil sample by culturing in nutrient medium with amylolytic activity 12.17 U/ml and cyclodextrinogenic activity up to 0.530 U/ml. Cyclodextrin glucanotransferase isolated from B. circulans B-65 shows the high degree for conversion of starch to cyclodextrins and this enzyme is specific for formation of β-cyclodextrin. Invention can be used in food industry for preparing cyclodextrins and cyclodextrin glucanotransferase used in different branches of industry.

EFFECT: improved isolating method, valuable properties of strain and enzymes.

9 cl, 8 tbl, 2 ex

The invention relates to biotechnology and is designed to produce carbohydrates fokusirovannyi

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. Disclosed is a method of producing β-cyclodextrin. The method includes preparing a suspension of maltodextrin, carrying out a ring closure process using a CGTase enzyme preparation in the presence of a trichloroethylene complexant. Further, the method includes removing the complexant by boiling, purifying the obtained product with activated carbon, separating the activated carbon by filtering and evaporating the syrup and crystallisation thereof while stirring and lowering the temperature. Finally, the method includes centrifuging and washing β-cyclodextrin crystals with cold water, drying and packaging.

EFFECT: invention can be used to produce cyclic complexing agents, particularly β-cyclodextrin.

2 dwg

FIELD: chemistry.

SUBSTANCE: group of inventions relates to biotechnology, particularly to mutant O-phosphoserine sulfhydrylase (OPSS) from Mycobacterium smegmatis with an amino acid sequence corresponding to SEQ ID NO: 1 which is devoid of three to seven C-terminal amino acid residues. The inventions also relate to a nucleic acid molecule encoding the mutant OPSS, an expression vector carrying the nucleic acid molecule, and a transformant transformed by the expression vector. In addition, a method is provided for producing cysteine in which O-phospho-L-serine (OPS) is reacted with a sulphide in the presence of the mutant OPSS.

EFFECT: muntant OPSS has improved enzymatic activity and can be used for environmentally friendly production of L-cysteine in conditions through a simple enzymatic conversion reaction; the group of inventions provides high output of L-cysteine.

16 cl, 7 dwg, 20 tbl, 19 ex

FIELD: chemistry.

SUBSTANCE: invention relates to the field of biotechnology. Claimed are version proteins of lysophospholipid-acyltransferase, catalysing a reaction of converting 18:3(n-6)-PL into 18:3(n-6)-CoA and/or DGLA-CoA into DGLA-PL, which have amino acid sequences, at least, by 95% identical to SEQ ID NO:2 and 7, given in the description. In addition, nucleic acids, coding the said proteins, are disclosed. Also claimed are: a recombinant expression vector, containing the said nucleic acids, and a host cell for obtaining a composition of fatty acids, transformed with the said vector. Claimed is the application of the said vector for increasing a quantitative ratio of arachidonic acid (ARA) in the composition of fatty acids in the host. Described is a method of obtaining the composition of fatty acids, which includes cultivation of the host cell for obtaining the composition of fatty acids with the increased quantity of arachidonic acid in comparison with its quantity in the composition of fatty acids, obtained by cultivation of a non-transformed host, and the collection of the composition of fatty acids from cultures of the transformed cell.

EFFECT: invention makes it possible to obtain the composition of fatty acids with an increased content of arachidonic acid.

18 cl, 8 dwg, 8 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method for determination and quantitative estimation of unusually modified glycans, which can be used in analysing glycans. The disclosed method includes the following steps: a) providing a glycan preparation containing unusually modified glycans selected from a group containing sulphated glycans, phosphorylated glycans, polyacetylated sialylated glycans and combinations thereof, where said unusually modified glycans are negatively charged and where said glycan preparation is obtained by separating glycans and sialic acids from a therapeutic glycoprotein composition, where the sialic acids are separated by exposing the therapeutic glycoprotein composition to at least one agent which splits sialic acid residues in conditions which facilitate splitting of sialic acids; b) subjecting the glycan preparation to a chromatographic separation method which separates glycans based on the charge-to-mass ratio, thereby separating said unusually modified glycans; and c) quantitative determination of at least one separated unusually modified glycan using at least one quantitative estimation standard.

EFFECT: novel efficient method which enables to analyse unusually modified glycans.

42 cl, 7 dwg, 2 tbl, 1 ex

Proteins // 2518345

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology, particularly to a version of the enzyme lipid acyltransferase with an increased phospholipid transferase activity. The prepared enzyme contains an amino acid motif GDSX, wherein X represents one or more of the following amino acid residues L, A, V, I, F, Y, H, Q, T, N, M or S and a modification of one or more amino acids as compared to the reference enzyme. The given enzyme is used for preparing food stuff, as well as for reducing the content of phospholipids in edible oil.

EFFECT: invention enables providing the effective version of the enzyme with high phospholipid transferase activity.

8 cl, 61 dwg, 1 tbl, 10 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology and discloses novel diacylglycerol acyltransferase. The invention also relates to a polynucleotide which codes diacylglycerol acyltransferase, an expression vector and a transformant such as yeast or fungus, as well as a method of preparing a composition of lipids or fatty acids using the transformant.

EFFECT: invention enables to obtain a novel enzyme which is suitable for efficient production of lipids and fatty acids.

13 cl, 5 dwg, 2 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: group of inventions relates to biotechnology. Disclosed is a composition for producing a fragrant ester. The composition contains SGNH-acyltranferase, an alcohol substrate containing 2-10 carbon atoms, and an acyl donor which is an ester substrate containing an acyl chain consisting of 2-10 carbon atoms. The alcohol substrate and the acyl donor are selected such that they produce a fragrant ester. Also disclosed is a method of producing a fragrant ester, according to which SGNH-acyltransferase, the alcohol substrate and acyl donor are combined in acyltransferase reaction conditions. Also disclosed is a method for simultaneous production of a bleaching agent, which is a peracid, and a fragrant ester. To this end, the SGNH-acyltransferase, alcohol substrate, acyl donor and aqueous hydrogen peroxide solution are combined in acyltransferase reaction conditions. The disclosed composition is used in detergents for removing stains, which contain at least one triglyceride, and for reducing unpleasant smells.

EFFECT: said SGNH-acyltransferase catalyses transfer of the acyl group from the acyl donor to the alcohol substrate to form a fragrant ester in an aqueous medium.

19 cl, 18 dwg, 9 tbl, 14 ex

FIELD: biotechnologies.

SUBSTANCE: new genes of lysophosphatidic acid acyltransferase were obtained from filamentous fungus Mortierella alpina. Nucleotide sequence coding enzyme was determined, structuring of expression vectors and vector construct by host cell creation containing it was described, and recombinant forms of lysophosphatidic acid acyltransferase coded by new genes were obtained.

EFFECT: higher efficiency.

4 cl, 14 tbl, 6 dwg, 9 ex

FIELD: biotechnologies.

SUBSTANCE: invention proposes use of a composition containing transglycosidase ferment (EC 2.4.1.24) for decomposition of polysaccharide of natural gum. The above polysaccharide of natural gum is a substrate for the above ferment of transglycosidase. A natural gum destruction method is described, which involves contact of transglycosidase ferment with polysaccharide of natural gum for destruction of the above polysaccharide of natural gum. Besides, a cleaning method is proposed, which involves contact of an object contaminated with polysaccharide of natural gum with cleaning composition including transglycosidase ferment; and maintenance of the above object and cleaning composition under conditions sufficient for effective destruction of polysaccharide of natural gum, and thus, cleaning of the above object.

EFFECT: invention allows decomposing natural gums by means of transglycosidase ferment.

19 cl, 7 dwg, 4 ex

FIELD: biotechnologies.

SUBSTANCE: method for inhibition of RNA-polymerase ferment activity is proposed. Inhibition is performed by introduction to a transcriptional system of an inhibitor based on at least one compound containing organic macrocyclic complex with encapsulated ion of transition metal, which has been chosen from the group including 1,8-bis(2-fluorobora)-2,7,9,14,15,20-hexaoxa-3,6,10,13,16,19-hexaasa-4,5,11,12-tetraphenyl-17,18-diamino-bicyclo[6.6.6]eicosa-3,5,10,12,16,18-hexaen(2-) ferrum(2+); 1,8-bis(2-fluorobora)-2,7,9,14,15,20-hexaoxa-3,6,10,13,16,19-hexaasa-4,5,11,12-tetraphenyl-17,18-dimethyl-bicyclo[6.6.6]eicosa-3,5,10,12,16,18-hexaen(2-) cobalt(2+); 1,8-bis(2-phenylbora)-2,7,9,14,15,20-hexaoxa-3,6,10,13,16,19-hexaasa-4,5,11,12,17,18-hexa(methyl thio)-bicyclo[6.6.6]eicosa-3,5,10,12,16,18-hexaen(2-) ruthenium(2+); 1,8-bis(2-fluorobora)-2,7,9,14,15,20-hexaoxa-3,6,10,13,16,19-hexaasa-4,5,11,12-tetraphenyl-17,18-cyclohexanedimercaptylbicyclo[6.6.6]eicosea-3,5,10,12,16,18-hexaen(2-) ferrum(2+); 1,12-bis-(tert-butylbora)-2,11,13,22,23,32-hexaoxa-3,10,14,21,24,31-hexasapentacyclo[11,11.11.04.9015,20025.30]ditri-akonta-3,9,14,20,24,30-hexaen(2-) ferrum(2+).

EFFECT: method provides for inhibition of activity of RNA-polymerase at micromolar concentration of inhibitor and enlarges the range of inhibitors of RNA-polymerase.

22 ex

FIELD: biotechnology, biochemistry, microbiology.

SUBSTANCE: the strain Bacillus circulans B-65 a producer of cyclodextrin glucanotransferase is isolated and selected form the soil sample by culturing in nutrient medium with amylolytic activity 12.17 U/ml and cyclodextrinogenic activity up to 0.530 U/ml. Cyclodextrin glucanotransferase isolated from B. circulans B-65 shows the high degree for conversion of starch to cyclodextrins and this enzyme is specific for formation of β-cyclodextrin. Invention can be used in food industry for preparing cyclodextrins and cyclodextrin glucanotransferase used in different branches of industry.

EFFECT: improved isolating method, valuable properties of strain and enzymes.

9 cl, 8 tbl, 2 ex

FIELD: biotechnology, in particular method for construction and production of mutant transglutaminases (MTG).

SUBSTANCE: invention relates to method for construction and production of mutant transglutaminases based on space structure of transglutaminase obtained from Streptoverticillium mobaraense, as well as to mutant MTG obtained by said method. Also disclosed are method for MTG modification based on space structure and modified transglutaminase with enhanced reactivity relative to substrate. Method of present invention makes it possible to predict MTG binding site to substrate on the base of space structure that is determined by MNG crystal X-ray analysis, and to design mutant transglutaminases by replacement, insertion or deletion of amino acid residues disposed on transglutaminase substrate-binding site.

EFFECT: new method for production and modification of mutant transglutaminases.

6 cl, 60 dwg, 5 ex

FIELD: biotechnology, peptides, genetic engineering.

SUBSTANCE: invention relates to constructing nucleic acid molecule comprising functional in starch-containing plant tissues promoter, fragment encoding transit peptide for translocation of useful peptide into amyloplast, fragment encoding useful peptide, region encapsulating into starch and terminator. In insertion into plant genome DNA molecule expresses hybrid polypeptide comprising the desirable protein encapsulated into starch matrix. Prepared vegetable material can be used, for example, in manufacturing fodders for mammals, fishes and poultries. Also, invention can be used in food industry.

EFFECT: valuable properties of nucleic acid and polypeptides.

15 cl, 19 dwg, 9 tbl, 7 ex

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