9268results about "Hydrolases" patented technology

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 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 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 discloses a rational copolymerization strategy for improving the stability of an oligomeric structure of acid-induced high-order oligomeric decarboxylase AdiA in a neutral to alkaline environment and application of the rational copolymerization strategy, and belongs to the field of bioengineering. According to the strategy, rational charge overturning transformation is carried out on a pentamer radial oligomeric interface and a dimer weft-wise oligomeric interface at the same time, and stable assembly and efficient catalysis of a decamer under the condition that the pH value is from 7.0 to 9.0 from neutral to alkaline are achieved. 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 AdiAD471K / E467K / H736E is 45.5 times that of a wild type when the pH value is 8.0, and the representative double mutant AdiAD471K / E467K / H736E keeps a complete decamer state when the pH value is 7.0-9.0. The yield of butanediamine is up to 156.5 g / L by using the mutant to catalyze whole cells.

Systems and methods for targeted gene modification, targeted insertion, perturbation of gene transcripts, and nucleic acid editing. Novel nucleic acid targeting systems comprise components of CRISPR systems and non-LTR retrotransposon elements.

The invention discloses SGNH family esterase SH2 screened in a Tibetan hot spring environment and application of the SGNH family esterase SH2, and belongs to the technical field of gene engineering. According to the invention, a new SGNH family esterase gene is screened from a Tibetan hot spring environment, and recombinant expression is realized in escherichia coli. The recombinant esterase prepared by the invention has the characteristics of thermal stability, salt resistance, organic solvent resistance and the like, can be safely applied to tooth whitening or skin exfoliating products, avoids damage of traditional chemical bleaching agents to enamel, and has important application value in the fields of daily chemicals, medicines, food processing and the like.

The invention discloses protease for improving the resistance of potatoes to late blight as well as a coding gene and application thereof, and belongs to the technical field of genetic engineering and plant disease resistance breeding. The amino acid sequence of the protease StRD21 is as shown in SEQ ID NO.2, or the protease StRD21 has a derivative sequence with the same disease-resistant function. The nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention also provides a specific primer pair for cloning the gene, a potato recombinant overexpression vector containing the gene and a transgenic host cell. The StRD21 gene is introduced into potatoes and overexpressed, so that the resistance of the potatoes to late blight can be remarkably enhanced. The invention provides an effective gene resource and a biotechnological means for cultivating a new variety of disease-resistant potatoes.

Methods for producing a linear deoxyribonucleic acid (DNA) product with enhanced resistance to nuclease digestion are provided. The methods comprise, (a) digesting a double-stranded DNA molecule with an endonuclease that cleaves an endonuclease target sequence to generate a digested double-stranded DNA molecule, wherein the digested double-stranded DNA molecule comprises a linear double-stranded region, and a truncated protelomerase sequence at a first end, wherein the truncated protelomerase target sequence is non-functional; (b) appending a first adaptor molecule to the first end of the digested double-stranded DNA molecule and appending a second adaptor molecule to the second end of the digested double-stranded DNA molecule to generate a precursor double-stranded DNA molecule, wherein the first adaptor molecule comprises a truncated protelomerase target sequence that forms a first functional protelomerase target sequence with the truncated protelomerase sequence at the first end of the digested double-stranded DNA molecule; and (c) incubating the precursor double-stranded DNA molecule with a protelomerase to generate the linear DNA product, wherein the protelomerase closes the first end of the precursor double-stranded DNA molecule at the first functional protelomerase target sequence.

The present invention relates to a nuclease polypeptide, such as a CRISPR nuclease polypeptide derived from a reference nuclease, which may be nuclease A, nuclease K or nuclease M, comprising a RuvC nuclease domain and a HNH nuclease domain. Also provided herein are gene editing systems comprising such nuclease polypeptides and gene editing methods using the gene editing systems.

The invention discloses a rice salt stress resistant gene OsHAK11 coding protein and application thereof, and belongs to the field of plant genetic engineering. According to the method, a rice OsHAK11 gene (the nucleotide sequence is shown as SEQ ID NO.1) is knocked out through a CRISPR / Cas9 gene editing technology, and a mutant with significantly enhanced salt tolerance is obtained. The gene editing vector pEGCas9Pubi-B-OsHAK11 contains sgRNA of a region as shown in a target SEQ ID NO.4, a rice receptor material is transformed through agrobacterium tumefaciens mediation, and a plant with the OsHAK11 gene subjected to frame shift mutation is obtained through screening. A salt stress experiment shows that the survival rate of the mutant oshak11 is obviously higher than that of a wild type. The invention provides a new gene resource and an efficient technical means for salt-tolerant breeding of rice.

The invention discloses a small CRISPR (clustered regularly interspaced short palindromic repeats)-Cas gene editing system and application thereof. According to the invention, based on microbial genome and metagenome data, a class of CRISPR-Cas family protein is mined through a biological information method, and is named as Cas12r. A CRISPR-Cas12r editing tool constructed on the basis of the gene can realize gene editing in prokaryotic or eukaryotic cells. The CRISPR-Cas12r gene editing system obtained by the invention has the characteristics of miniaturization and various PAM types.

The disclosure encompasses genetically modified plants and methods for improving phosphorus redistribution and use efficiency. The genetically modified plants increase the expression of a nonspecific phospholipase C4 (NPC4) protein, enhancing phosphorus remobilization from old, senescing tissues to young, growing tissues and seeds. The genetic modification improves plant growth, increases seed and oil yield, and alters fatty acid composition. The genetically modified plants demonstrate enhanced phosphorus use efficiency under phosphorus-limited conditions, showing greater growth and yield improvements compared to wild-type plants. Specifically in camelina plants, the genetic modification increases seed yield, elevates oil content, and alters fatty acid composition of the oil under P-limited conditions.

The invention discloses a D-psicose 6-phosphate phosphatase mutant and application of the D-psicose 6-phosphate phosphatase mutant in preparation of psicose, and belongs to the technical field of enzyme engineering. According to the invention, site-directed mutagenesis is carried out on an amino acid sequence of D-psicose 6-phosphate phosphatase A6PP, and a combined mutant I21V / Y73F / R115M / T180V / L203I / Q193E is constructed, so that the substrate specificity, the thermal stability and the catalytic activity of the combined mutant are all remarkably improved. The invention further provides application of the combined mutant in preparation of psicose through multi-enzyme cascade catalysis, 150 g / L of maltodextrin can be converted into 97.7 g / L of D-psicose, the conversion rate reaches 65.1%, and the application prospect of the combined mutant in the field of food is widened.

The invention discloses a primer group, a kit and a method for detecting brucella, and belongs to the technical field of gene detection. The primer group comprises two pairs of specific RPA (recombinase polymerase amplification) primers: a primer pair B1F1 (SEQ ID No.3) and a primer pair B1R1 (SEQ ID No.6) targeting a B1 sequence, and a primer pair B2F1 (SEQ ID No.8) and a primer pair B2R1 (SEQ ID No.11) targeting a B2 sequence. The invention also provides a kit containing the primer group, and a detection method based on the RPA-CRISPR-Cas12a. The invention also provides a kit containing the primer group and a detection method based on the RPA-CRISPR-Cas12a. According to the application, the high efficiency of RPA amplification is combined with the ultrahigh specificity of CRISPR-Cas12a detection, the defects of long time consumption, low sensitivity and easy generation of false positive in the traditional method are overcome, rapid, sensitive and specific detection of Brucella is realized, and the application has important application value in early screening and monitoring of Brucella.

The disclosure provides, for example, double stranded DNA (dsDNA) molecules comprising one or more chemically modified nucleobases. In some embodiments, the dsDNA molecule is circular and comprises a first strand and a second strand, wherein the first strand comprises one or more chemically modified nucleobases, and the second strand is free of chemically modified nucleobases. In some embodiments, the dsDNA molecule comprises a promoter sequence and an effector sequence that encodes an effector.

Disclosed herein are methods of increasing nitrogen fixation in a non-leguminous plant. The methods can comprise exposing the plant to a plurality of bacteria. Each member of the plurality comprises one or more genetic variations introduced into one or more genes or non-coding polynucleotides of the bacteria's nitrogen fixation or assimilation genetic regulatory network, such that the bacteria are capable of fixing atmospheric nitrogen in the presence of exogenous nitrogen. The bacteria are not intergeneric microorganisms. Additionally, the bacteria, in planta, produce 1% or more of the fixed nitrogen in the plant.

Provided are variant Cas12f4 polypeptides, fusion proteins containing variant Cas12f4 polypeptides, nucleic acids encoding variant Cas12f4 polypeptides and fusion proteins, compositions and methods related to those variant Cas12f4 polypeptides and fusion proteins, and modified host cells comprising the variant Cas12f4 polypeptides and / or encoding nucleic acids. The variant Cas12f4 polypeptides disclosed herein exhibit improved functionality, stability, and / or other desirable properties and may be used in combination with guide Cas12f4 RNAs that can be used in a variety of applications ex vivo, in vitro, and in vivo.

The application relates to an engineering bacterium for high yield of observation blue and a construction method and application thereof, and belongs to the technical field of biological synthesis of natural dyes. The engineering bacterium for high yield of observation blue expresses icd, bpsA, glnA Y405F and gdhA; the engineering bacterium knocks out acnR, yggB, glsK, aceA and ldh. By knocking out the yggB and aceA genes, performing site-directed mutation (Y405F) on the glnA gene, replacing the glsK gene with the glnA Y405F gene, replacing the acnR gene with the icd gene, replacing the ldh gene with the gdhA gene, and integrating the bpsA gene, the application can block the formation of by-products such as lactate and succinic acid in the observation blue synthesis process, improve the flow direction of the citric acid->isocitric acid->alpha-ketoglutaric acid->glutamic acid->glutamine path, and then improve the intracellular glutamine concentration, so that the yield of observation blue is improved.

The invention discloses enterobacter hormaechei XW-01 and a microbial preparation thereof, the strain (preservation number is CCTCC NO: M 20242595) can efficiently degrade protein, total sugar and nicotine in tobacco leaves, sensory indexes such as aroma and aroma quantity are remarkably improved, and protease activity is remarkably improved when the enterobacter hormaechei XW-01 and wheat bran act synergistically. The microbial preparation is obtained through the processes of strain activation, fermentation culture and microbial agent preparation, and when the microbial preparation is applied to tobacco leaf treatment, through the steps of water content adjustment, preparation spraying, sealed fermentation, low-temperature drying and the like, the technical breakthrough that the protein content is reduced, the tar release amount is reduced, and the comprehensive quality grade is improved is achieved. Compared with a traditional method, the technology has the advantages that the treatment period is shortened, the cost is greatly reduced, the COD value of wastewater is greatly reduced, the cigarette quality and the production environmental protection property are remarkably improved, and the method has a wide industrial application prospect.

The present disclosure relates to compositions and methods of increasing levels of fetal hemoglobin (HbF) in cells. The present disclosure further relates to methods for treating patients suffering from blood cell diseases, including those associated with reduced amounts of functional adult hemoglobin (HbA), such as sickle cell disease and β-thalassemias

Disclosed herein are an engineered immune cell with CD7 gene knock-out and use thereof. According to the present invention, an sgRNA specifically targeting CD7 gene is designed and synthesized, which can accurately target CD7 gene to achieve gene knock-out with high knock-out efficiency. The provided sgRNA can be used for preparing a CD7-targeting engineered immune cell, and can be further used for preparing a CD7-targeting universal CAR-T cell

The invention provides a structure, a sequence and a construction method of a plant virus gene editing vector and application of the plant virus gene editing vector in plant gene editing. The method is characterized in that a TRV2 genome of a tobacco embrittlement virus (TRV) is used for carrying and expressing a small Cas gene AsCas12f and gRNA of the small Cas gene AsCas12f. Preferably, the AsCas12f and the gRNA of the AsCas12f are subjected to tandem expression by utilizing the TRV2 to form an expression cassette structure of the AsCas12f-tRNA-gRNA, the expression cassette substitutes a part of 2b gene sequence of the TRV2 on a TRV2 genome to form a plant gene editing vector pTRV2-Cas12f-gRNA based on the TRV virus, and long-time stable expression and gene editing can be performed after wild type nicotiana benthamiana is inoculated. The invention also provides a method for carrying out gene editing in a wild type plant by utilizing the plant virus.

The present invention relates to modified Dda helicases which can be used to control the movement of analytes such as polynucleotides. The modified Dda helicases are used in analyte detection and characterisation. The present invention also relates to novel protein pores and their uses in analyte detection and characterisation. The invention particularly relates to an isolated pore complex formed by a CsgG-like pore and a modified CsgF peptide, or a homologue or mutant thereof, thereby incorporating an additional channel constriction or reader head in the nanopore.

The invention belongs to the technical field of genes, and discloses a method for efficiently expressing a foreign protein based on an NC048604-1 site in a CHO cell genome. A site for stably expressing protein in the CHO cell genome is located in the 94142000 to 94148000 basic group range of the CHO cell genome NC048604-1, and the nucleotide sequence of the site is as shown in SEQ ID NO: 1. According to the invention, different protein genes are introduced at fixed positions in a CHO cell genome, and stable expression is carried out.

The invention provides an application of WNT7B in construction of a myopia animal model. Homologous genes wnt7ba and wnt7bb of the WNT7B in the zebra fish are knocked out or knocked down by applying a CRISPR / Cas9 gene editing technology, an animal model of which the eye axis length is remarkably increased, the eyeball movement frequency is remarkably reduced and the light response is reduced is obtained, and the result shows that the WNT7B is related to the high myopia. The model provides a favorable tool for pathogenesis and drug screening of high myopia, and has a good application prospect.

The invention provides sgRNA for targeting and guiding nuclease to efficiently cut an HLA-DRA gene, a method for modifying a CAR-T cell by using the sgRNA, and a related gene editing system, reagent and kit. According to the CAR-T cell, the immunogenicity is greatly reduced, the risk of graft versus host disease and immunological rejection can be effectively reduced, and meanwhile the killing capacity of tumor cells of the CAR-T cell is not affected.

Provided are a marine phospholipase C mutant and a use thereof. Compared with wild-type (WT) enzyme protein, at 40°C and under conditions where enzyme protein activity remains essentially unchanged, the enzyme protein half-life of the constructed mutant can be increased to 3.46 and 9.25 times that of the original enzyme protein, and the residual phosphorus content in degummed oil can be reduced from 551 ppm to 72 ppm, which shows a significant improvement over the final residual phosphorus content of 175 ppm for the blank control and the final residual phosphorus content of 131 ppm for the WT. These results demonstrate that the enzyme has improved thermal stability, enables oil degumming under various conditions, and is suitable for a broader range of industrial applications.

The invention belongs to the technical field of bioengineering, and particularly relates to a zearalenone toxin degrading enzyme variant and application thereof. The amino acid sequence of the zearalenone toxin degrading enzyme is as shown in SEQ ID NO.2, or the zearalenone toxin degrading enzyme with the amino acid sequence as shown in SEQ ID NO.2 is obtained through amino acid mutation. The zearalenone toxin degrading enzyme mutant is obtained through mutation screening, and compared with a wild type, the zearalenone toxin degrading enzyme mutant has the advantages that the zearalenone degrading capability is obviously improved; besides, the zearalenone toxin degrading enzyme is expressed by using alfalfa, the zearalenone toxin degrading enzyme with biological activity is easy to obtain, and the zearalenone toxin degrading enzyme has certain application potential in prevention and treatment of animal poisoning caused by zearalenone toxin pollution in agriculture and animal husbandry production.

The invention provides sgRNA for targeting and guiding nuclease to efficiently cut a B2M gene, a method for modifying cells by using the sgRNA, and a related gene editing system, reagent and kit. The cells produced by the invention greatly reduce the immunogenicity, can effectively reduce the risk of graft versus host disease and immunological rejection, and does not affect the killing ability of the cells to tumor cells.

Provided herein are engineered immune cells and populations thereof for administration to patients to treat cancer (e.g., solid tumors or liquid tumors) and other conditions. The cells are engineered to functionally express a reduced level of one or more of RFX5, NLRC5, TAP2, β2m, TRAC, RFXAP, CIITA and RFXANK. The cells optionally are further engineered to express one or more than one additional protein such as an antigen binding protein (e.g., a chimeric antigen receptor (CAR) or T cell receptor) to target tumor cells or other damaged cells in the patient and / or to express other genes at a reduced level. Also provided are methods of making and using the engineered cells, compositions and kits comprising them, and methods of treating by administering the cells and the compositions.