18138results about "Hydrolases" patented technology

Described herein, inter alia, are GTPase inhibitors and uses thereof.

Provided herein are methods, compositions, and kits for in vitro transcription of spatially captured target nucleic acids on an array.

The invention provides structure-oriented excavated cytosine deaminase with high editing activity and no sequence preference and application of the structure-oriented excavated cytosine deaminase. The invention discloses a series of novel cytosine deaminase. The cytosine deaminase disclosed by the invention has enhanced targeted editing performance; the enhanced targeted editing performance includes improved sequence-free preference, improved editing efficiency, reduced off-target effect, and improved targeted specificity.

The invention relates to an engineering bacterium of high-yield ornamental blue as well as a construction method and application of the engineering bacterium, and belongs to the technical field of natural dye synthesis by a biological method. The engineering bacterium capable of highly producing the ornamental blue, disclosed by the invention, expresses icd, bpsA, glnAY405F and gd hA; acnR, yggB, glsK, aceA and ldh are knocked out from the engineering bacteria. According to the application, yggB and aceA genes are knocked out, site-directed mutagenesis (Y405F) is carried out on a glnA gene, a glsK gene is replaced by a glnAY405F gene, an acnR gene is replaced by an icd gene, an ldh gene is replaced by a gdhA gene, and a bpsA gene is integrated, so that the formation of by-products such as lactate and succinic acid in a blue-viewing synthesis process can be blocked; the flow direction of a passage from citric acid to isocitric acid to alpha-ketoglutaric acid to glutamic acid to glutamine is improved, so that the concentration of glutamine in cells is improved, and the yield of the ornamental blue is improved.

The invention discloses a biosynthesis system for high-yield 1, 6-hexamethylenediamine and application of the biosynthesis system, and belongs to the technical field of biology. According to the method, key genes which are excavated and optimized in a 1, 6-hexamethylenediamine biosynthetic pathway are co-expressed in a single cell module or a multi-cell module, and different enzyme elements expressed by the cell module are subjected to enzyme cascade reaction, so that 1, 6-hexamethylenediamine can be efficiently synthesized by taking caprolactam or 6-aminocaproic acid as a substrate; furthermore, after the carboxylic acid reductase MaCAR is mutated, the catalytic activity of the carboxylic acid reductase MaCAR can be remarkably improved, and the yield of the 1, 6-hexamethylenediamine is further improved; in addition, the biosynthesis system disclosed by the invention directly takes the cell module as a whole-cell catalyst, and an enzyme purification step is avoided, so that the biosynthesis system has the advantage of low cost.

The invention relates to a high-yield lipase strain and application thereof, and belongs to the technical field of bioengineering. According to the invention, grease-rich samples from different sources are subjected to enrichment treatment, a strain with lipase activity reaching 80.8 + / -1.8 U / mL is obtained through neutral red grease plate screening, the strain is identified as Burkholderia cepacia OCRI-Lip100 through sequencing and sequence comparison, the Burkholderia cepacia OCRI-Lip100 is named as Burkholderia cepacia OCRI-Lip100, and the Burkholderia cepacia OCRI-Lip100 is preserved in China Center for Type Culture Collection on July 3, 2024 with a preservation number of CCTCC M20241369. Lipase obtained by fermentation of the strain is sequenced, enzymatic properties, catalytic activity and substrate preference of the lipase are studied, the optimum catalytic temperature is 60 DEG C, the specific enzyme activity is higher than that of commercial lipase, the catalytic activity is further improved to 116 + / -0.5 U / mL when a C14 substrate is catalyzed, and triglyceride can be hydrolyzed into diglyceride. The strain provided by the invention and the lipase produced by the strain enrich the resource library of the existing lipase, and provide a basis for improving the industrial application of the lipase and realizing the high utilization rate of resources.

The present invention provides a method, application and product for preparing diglyceride with weight loss effect by enzymolysis, which belongs to the field of enzyme engineering technology. The method comprises the following steps: S1, mixing lipase, ammonium heptamolybdate tetrahydrate with water, adding glycerol precipitation, filtering, and obtaining an enzyme preparation; S2, mixing plant raw material oil, glycerol, fatty acid, sodium fatty acid with the enzyme preparation obtained in step S1, reacting under inert gas conditions, and obtaining an enzymolysate; S3, molecularly distilling the enzymolysate obtained in step S2 to obtain diglyceride; wherein the lipase is a mixture of Rhizomucor miehei lipase and Candida antarctica Aspergillus niger lipase. The present invention uses ammonium heptamolybdate tetrahydrate to load a specific lipase mixture, and at the same time optimizes the component types and ratios of the raw materials in the enzymolysis process, effectively improving the mass percentage content and diglyceride yield of enzymolysis production of diglyceride.

Among other things, the present disclosure provides oligonucleotide compositions and methods thereof. In some embodiments, the present disclosure provides structural elements including various chemical modifications of sugars, nucleobases and / or internucleotidic linkages that are useful for adenosine editing. In some embodiments, the present disclosure provides methods for preventing or treating conditions, disorders or diseases that can benefit from editing of an adenosine in a nucleic acid.

The invention relates to the technical field of biology, and discloses yarrowia lipolytica engineering bacteria with high yield of p-coumaric acid by taking glucose as a substrate as well as a construction method and an application of the yarrowia lipolytica engineering bacteria. The construction method comprises the following steps: integrating a tyrosine ammonolysis enzyme gene TAL into a Yarrowia lipolytica strain by adopting a CRISPR / Cas9 positioning integration method, strengthening DAHP synthetase genes ARO4, DHS1 and AROG, overexpressing a tyrosine synthetase TYR gene and a histidine phosphate aminotransferase HIS5 protease gene, and carrying out high-throughput screening to obtain the Yarrowia lipolytica strain. And further integrating exogenous phenylalanine deamination hydroxylation pathway genes for synthesizing p-coumaric acid, wherein the exogenous phenylalanine deamination hydroxylation pathway genes comprise a phenylalanine ammonialyase gene, a cinnamate hydroxylase gene and a P450 reductase gene. The yield of p-coumaric acid produced by the Yarrowia lipolytica engineering strain is the highest yield of known Yarrowia lipolytica, the highest yield in a shake flask can reach 1.7 g / L, the fed-batch fermentation yield in a 5L fermentation tank can reach 30 g / L, and the Yarrowia lipolytica engineering strain has remarkable industrial application value.

The invention provides a Pseudomonas protegens P150 strain and an application of the Pseudomonas protegens P150 strain, and belongs to the technical field of biocontrol bacteria. The defensive pseudomonas P150 is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.34520, the preservation time is May 13, 2025, and the preservation address is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The defensive pseudomonas P150 disclosed by the invention has the effects of inhibiting various pathogenic bacteria, producing enzymes, promoting growth and preventing and treating tomato bacterial wilt and soybean pustule. The microbial inoculum prepared by adopting the biocontrol bacterium can reduce the dosage of chemical pesticides and the prevention and control cost of plant diseases, is green and pollution-free, can be used for preventing and controlling tomato bacterial wilt and soybean pustule, and has a relatively good popularization value.

The invention relates to a saccharomyces cerevisiae strain for efficiently synthesizing cordycepin as well as a construction method and application of the saccharomyces cerevisiae strain, and belongs to the technical field of microbial genetic engineering. The construction method provided by the invention comprises the following steps: carrying out multi-copy genome integrated expression on heterologous codon optimized 2 '-carbonyl-3'-deoxyadenosine reductase genes and 3 '-adenosine monophosphate phosphohydrolase genes, so as to increase the dosage of key enzymes; and supplementing the defective gene of the engineering strain to obtain the saccharomyces cerevisiae strain B5U3 with high yield of cordycepin. According to the obtained saccharomyces cerevisiae strain for efficiently synthesizing the cordycepin, the cordycepin yield reaches 2.92 g / L after shake flask fermentation, the cordycepin yield reaches 7.5 g / L after fermentation in a fermentation tank, and a foundation is laid for safer and more reliable industrial production of the cordycepin.

Disclosed are compositions of lipid nanoparticles (LNP) comprising an ionizable cationic lipid, a phospholipid, a sterol, and a PEG-lipid (non-functionalized and optionally functionalized). The functionalized PEG-lipid can be conjugated with a binding moiety to create a targeted LNP (tLNP). The disclosed tLNP preferentially deliver a nucleic acid molecule or other negatively charged payload to cells expressing a cell surface antigen recognized by the binding moiety of the tLNP, and are better tolerated, as compared to LNPs and tLNPs comprising ionizable cationic lipids found in marketed pharmaceuticals comprising LNPs.

The invention relates to the technical field of bioengineering, and discloses a mutant of Burkholderia lipase as well as a recombinant expression vector and application of the mutant, the mutant comprises P131S, T132A, T251A, T251S, P131A, P131Q, T132G, Q88S and P113A single processes; and a Q88S double process and a P113A double process; and a multi-site mutation combination: a combination I: Q88S, P113A, T132G and L287S, and a combination II: Q88S, P113A, T132G and L287S; a combination II: Q88S, P113A, P131S and L287S; and a combination III: Q88S, P113A, P131S and T251A. Based on experiments, the invention finds that each mutant can not only improve the protein yield, but also improve the enzyme activity by 57% at most; ePA enrichment experiments find that the EPA content of the mutant can be increased by 33% at most, and the mutant has excellent EPA purification and enrichment effects.

The present disclosure provides compositions and methods for prime editing with improved editing efficiency and / or reduced indel formation with modified prime editors and prime editor fusion proteins. The disclosure further provides, vectors, cells, and kits comprising the compositions and polynucleotides of the disclosure.

The invention provides a modified TnpB-omega RNA gene editing system and application of the modified TnpB-omega RNA gene editing system. In particular, the present invention provides a modified TnpB related [omega] RNA skeleton. The modified omega RNA skeleton provided by the invention has at least three stem-loop structure fragments, and has a shorter length and a compact structure than a natural omega RNA skeleton, so that the modified omega RNA skeleton and TnpB can be jointly carried in a single AAV vector, the administration efficiency is improved, and high-efficiency gene editing is realized.

The present invention discloses a Gs12-7 endonuclease variant and a gene editing system mediated thereby. Specifically, by means of a rational mutation strategy, two Gs12-7 mutants were constructed and compared, and it was found that after the 157th amino acid of the Gs12-7 endonuclease was mutated from Glu to Arg, the activity of this gene editing enzyme could be significantly improved, and this variant was designated as Gs12-7MAX. The present invention provides an efficient gene editing technology mediated by the CRISPR-Gs12-7MAX system, which has broad application prospects in the field of genome site-directed modification.

Disclosed are compositions of lipid nanoparticles (LNP) comprising an ionizable cationic lipid, a phospholipid, a sterol, and a PEG-lipid (non-functionalized and optionally functionalized). The functionalized PEG-lipid can be conjugated with a binding moiety to create a targeted LNP (tLNP). The disclosed tLNP preferentially deliver a nucleic acid molecule or other negatively charged payload to cells expressing a cell surface antigen recognized by the binding moiety of the tLNP, and are better tolerated, as compared to LNPs and tLNPs comprising ionizable cationic lipids found in marketed pharmaceuticals comprising LNPs.

The invention relates to the technical field of biosynthesis, and particularly discloses a pyridone compound as well as a biosynthesis method and application thereof. On the basis of related technologies of synthetic biology, non-ribosome peptide synthetase is modified, non-ribosome peptide synthetase mutants are heterologously expressed in various hosts, a pyridone compound is found and obtained for the first time, the chemical molecular formula of the pyridone compound is determined to be C5H4N2O3, and the pyridone compound is named as 5-aminopyridine-2, 3, 6-triketone or 5-hydroxy-3-iminopyridine-2, 3, 6-triketone or 5-hydroxy-3-iminopyridine-2, 3, 6-triketone or 5-hydroxy-3-iminopyridine-2, 3, 6-triketone. The novel pyridone compound can be used as a raw material or an intermediate to be applied to development and preparation of novel antibiotics, drugs and dyes.

The invention discloses a sea cucumber protein heptapeptide with functions of reducing blood pressure and blood sugar and application of the sea cucumber protein heptapeptide, and belongs to the technical field of bioactive peptide biology. The amino acid sequence of the sea cucumber protein heptapeptide is LPPGPFP. The sea cucumber protein heptapeptide can significantly inhibit the activity of ACE through a competitive inhibition mode, tolerates the gastrointestinal tract digestion environment, also has DPP-IV inhibition activity, can relieve hypertension and type 2 diabetes mellitus at the same time, and has an application value in preparation of products with the efficacy of reducing blood pressure and blood sugar due to multiple functions of the sea cucumber protein heptapeptide. The sea cucumber protein heptapeptide provided by the invention can significantly inhibit the activity of ACE through a competitive inhibition mode, has DPP-IV inhibition activity, has the characteristics of simple structure, safety, strong activity and the like, and has a wide application prospect.

Provided are compositions for genome editing and site-directed integration in plants comprising microprojectile particles coated, treated of applied with a recombinant DNA construct comprising a sequence encoding one or more genome editing reagents for delivery to a mature embryo explant from dry seeds. Further provided are methods for genome editing and site-directed integration in at least one cell of a plant using the disclosed compositions, and plants, plant parts and seeds comprising an edited genome or site-directed integration, which are produced by the disclosed methods.

The invention discloses bacillus velezensis YB-1652, a biocontrol microbial inoculum and application of the biocontrol microbial inoculum, and aims to solve the technical problem of lack of effective biological control measures for continuous accumulation of peanut phenolic acid autotoxic substances and pathogens in continuous cropping. According to the application, a bacillus velezensis YB-1652 strain is separated and screened from rhizosphere soil of continuous cropping peanuts, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.36152. The strain has the capability of efficiently degrading phenolic acid compounds, relieves continuous cropping obstacles and improves the soil environment; the compound has a relatively strong inhibition effect on southern blight and root rot of peanuts, and the occurrence rate of soil-borne diseases is reduced; meanwhile, peanut growth can be promoted; the bacillus amyloliquefaciens has siderophore, cellulase, beta-glucanase, amylase power and protease production capacity. The YB-1652 strain and the preparation thereof can provide technical support for restoration of peanut continuous cropping obstacles, provide bacterial source guarantee for prevention and treatment of peanut soil-borne diseases, replace or reduce use of chemical pesticides and improve food and ecological environment safety, and have good economic and social benefits.

The invention relates to antisense oligonucleotides that can form a double stranded nucleic acid complex with a target RNA molecule, wherein the double stranded nucleic acid complex is capable of recruiting an adenosine deaminating enzyme for deamination of a target adenosine in the target RNA molecule, wherein the nucleotide in the AON that is directly opposite the target adenosine comprises a 2',2'-disubstitution, preferably a 2',2'-difluro substitution in the ribose moiety.

The invention discloses a Candida antarctica lipase B mutant with high transesterification activity and application thereof. The nucleotide sequence of the Candida antarctica lipase B mutant is shown as SEQ ID NO.1, and the amino acid sequence of the expression protein of the Candida antarctica lipase B mutant is shown as SEQ ID NO.2. The invention further discloses a preparation method of the Candida antarctica lipase B mutant. Compared with an original enzyme, the mutant has higher fat hydrolysis activity and transesterification activity, the specific enzyme activity of the fat hydrolysis activity reaches 480.39 U / mg, the specific enzyme activity of the transesterification activity reaches 40.67 U / mg, the biodiesel conversion rate at the temperature of 30 DEG C can reach 92%, low-temperature high catalytic activity is shown, and the mutant can be used for enzymatic preparation of biodiesel and has potential industrial application value.

The invention relates to the field of enzyme engineering, in particular to modified xylanase, a DNA (deoxyribonucleic acid) molecule, a recombinant plasmid and a transfected cell. The modified xylanase protected by the application is formed by replacing a deletion segment of a receptor enzyme with a substitution segment, the receptor enzyme is wild type xylanase, the sequence of the receptor enzyme is SEQ ID NO: 1, the deletion segment comprises a carbohydrate binding module of the receptor enzyme, the substitution segment is a carbohydrate binding module of a donor enzyme, the donor enzyme is a carbohydrate active enzyme, and the donor enzyme is a carbohydrate active enzyme. The sequence of the donor enzyme is one of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5. The carbohydrate binding module of the carbohydrate active enzyme with relatively high activity is replaced into the receptor enzyme, so that the binding capacity of the receptor enzyme and the carbohydrate is improved, and the activity of the receptor enzyme is improved.

The invention discloses an arginine decarboxylase diaA enzyme mutant and application thereof in preparation of butanediamine, and belongs to the field of bioengineering. According to the invention, rational charge overturning transformation is simultaneously carried out on a pentamer meridian oligomeric interface and a dimer latitudinal oligomeric interface, so that stable assembly and efficient catalysis of the decamer under the condition of neutral to alkaline pH (7.0-9.0) are realized. Wherein positive charges are introduced into a meridian interface to weaken electrostatic repulsion, and negative charges are introduced into a latitudinal interface to enhance dimer compactness and substrate transfer efficiency. The specific enzyme activity of the representative double mutant AdiAD110K / H736E at pH 8.0 is about 35 times that of a wild type, and the representative double mutant AdiAD110K / H736E keeps a complete decamer state in a pH range of 7.0-9.0. The yield of butanediamine is up to 145.9 g / L under the whole-cell catalysis of the mutant.

The invention relates to the technical field of food, in particular to diglyceride for reducing triglyceride, cholesterol and uric acid as well as a biological enzyme preparation method and application of the diglyceride. The preparation method comprises the following steps: (1) adding water and glycerol into vegetable oil, and then adding immobilized lipase to react to obtain a reactant; (2) centrifuging reactants, collecting a light-phase product, carrying out molecular distillation, and collecting a heavy-phase product to obtain diglyceride; the immobilized lipase comprises lipase and a carrier, the carrier comprises loofah sponge, soybean meal and bentonite, and the mass ratio of the loofah sponge to the soybean meal to the bentonite is 1: (2-4): (3-6). The method has the advantages that the effects of reducing triglyceride, total cholesterol and uric acid are relatively good, and the yield and the purity are relatively high.