112 results about "Enzyme protein" patented technology

The invention relates to the technical field of plant genetic engineering, and particularly provides cloning of a GbGELP25D gene of gossypium barbadense and application of the GbGELP25D gene of gossypium barbadense. According to the invention, the GbGELP25D gene with a full length of 1092 bp is cloned from the sea island cotton variety Xinhai No. 7 for the first time, and the gene encodes a secretory lipase protein with a signal peptide and is positioned in an extracellular gap. Through bioinformatics analysis, phylogenetic classification, protein structure modeling and signal peptide function verification, it is clear that the gene belongs to a plant GELP family. Furthermore, a virus-induced gene silencing technology is utilized to prove that the silent GbGELP25D can obviously enhance the resistance of cotton to verticillium wilt, and the mechanism of the silent GbGELP25D is closely related to activation of ethylene synthesis and signal channels and induction of expression of disease-resistant related genes. The resistance gene provided by the invention enriches gene resources of cotton verticillium wilt resistance breeding, and has important theoretical significance and application value.

This disclosure provides improvised reagents and methods for inserting a transgene into the genome of a living host cell. Sleeping beauty (SB) transposase is used in protein form rather than as a vector. This has been made possible by using rational mutagenesis in a particular region of the SB crystal structure to develop variant SB transposase protein that is more soluble. The changes increase solubility while maintaining transposase activity, thereby adapting the variant transposase for the purpose of promoting recombinant integration of a transgene into a target cell when used in protein form rather than as a polynucleotide vector. The modified transposase is highly soluble in electroporation buffer and thermostable during storage. When introduced into a host cell, it promotes integration of a transgene into the genome of the cell in a dose-dependent manner. It is degraded within 48 hours, thereby rapidly clearing transposase activity from the host cell.

The present application relates to a kind of cytochrome P450 oxidase CYP716C52 protein, and the CYP716C52 protein coding gene, the protein can catalyze hydroxylation of maytenic acid C2 position to generate triptolide acid C, in turn participate in triptolide biosynthesis.

The invention belongs to the technical field of biology, and provides an application of alpha-amylase protein derived from nephroplasmosis in culture of nephroplasmosis, and the alpha-amylase protein is obtained by synthesizing a gene sequence of alpha-amylase as shown in SEQ ID NO.2 after codon optimization and then transfecting the gene sequence to an expression vector. And carrying out bacterial transformation, culture, induction and crushing. The invention also provides a method for inducing the formation of the reproductive cyst of the reniform worm. It is found for the first time that alpha-amylase protein specifically induces reproductive cysts to be formed, the biomass of the nephroplasmosis is increased, the growth generation of the nephroplasmosis is shortened, and the method is suitable for species such as distended nephroplasmosis and nephroplasmosis henslouianum and fits natural ecological scenes; complex equipment is not needed, the cost is low, large-scale production is easy, the stability is high, and the method can be widely applied to soil ecological restoration, agricultural microbial resource development and basic research related to nephroplasmosis.

This invention pertains to fusion protein mutants comprising a Prime Editing enzyme having a first amino acid sequence and a second amino acid sequence, wherein the first amino acid sequence comprises a SpCas9 H840A nickase mutant protein of SEQ ID NO:152 and the second amino acid sequence comprises a Moloney Murine Leukemia Virus reverse transcriptase protein mutant (MMLV RTase mutant), wherein the fusion protein mutant displays at least the equivalent or greater activity of a reference Prime Editing enzyme in genome editing.

The present invention provides a method for producing a baked or par-baked product, comprising the steps of: a) preparing a dough comprising a mature thermostable variant of a parent glucoamylase that is at least 70% identical to SEQ ID NO: 1, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 10, added in an amount of 0.01 to 12.40 mg of enzyme protein (mgEP) per kg of flour, wherein the dough has a pH value in the range of 3.0 to 6.5; and b) baking or par-baking the dough to produce a baked or par-baked product.

The present application relates to the field of enzymatic reaction device, specifically to an enzyme protein ultrasonic pretreatment and temperature control enzyme acceleration reaction system and operation method, which is used to solve the contradiction between enzymatic efficiency and enzyme activity and the problem of insufficient dynamic stability of reaction conditions. The system includes a reaction container, a bearing structure, an ultrasonic transducer array, an ultrasonic generator, a temperature sensor, a temperature control device, an enzymatic reaction monitoring module and a controller; the ultrasonic pretreatment improves the accessibility of the substrate, the temperature control device maintains the stability of the reaction temperature, the monitoring module collects pH and turbidity data in real time, and the controller outputs a termination signal based on the turbidity threshold. The system is mainly used in the enzymatic reaction process of enzyme protein to improve the reaction efficiency and product quality.

The utility model relates to a device for removing impure protein in crude polysaccharide of traditional Chinese medicine by utilizing immobilized protease, which comprises a protein removal reactor loaded with the immobilized protease, a dissolved oxygen tester and a constant-temperature magnetic stirrer, and a dissolved oxygen electrode of the dissolved oxygen tester is connected to the protein removal reactor. The papain immobilized on the microcrystalline cellulose is used for carrying out enzymolysis reaction on impure protein in the traditional Chinese medicine crude polysaccharide, and the device has the characteristics of simplicity in operation, mild conditions and environment friendliness. The immobilized papain is limited between the two layers of screens, so that the problem of separation of the immobilized papain in the traditional Chinese medicine crude polysaccharide solution can be avoided, and the method has the characteristics of rapidness and convenience. The immobilized papain in the device can be expanded to immobilize other proteases, such as bromelain, subtilisin, protease K and the like, on cellulose according to protein types in different traditional Chinese medicine polysaccharide solutions.

The invention discloses a preparation method of composite aerogel, which comprises the following steps: stirring and mixing a silk fibroin solution and a methanol solution of Zn (CH3COO) 2.2 H2O, and dropwise adding a methanol solution of 2-methylimidazole for reaction to form a composite system; then adding polyvinyl alcohol, dissolving at high temperature, reacting at 60 DEG C for 0.5-2 hours, repeatedly freezing and unfreezing at-80 DEG C for 3-5 times, and finally freezing and drying to obtain the composite aerogel material. According to the invention, ZIF-8 is grown on silk fibroin in situ, abundant hydrophilic groups are provided by using a beta-folding structure of the ZIF-8, an active center and a zymoprotein environment of natural carbonic anhydrase are simulated, and proton transport and hydration reaction processes are promoted; meanwhile, a multistage pore channel is constructed by combining micropores of ZIF-8 and mesopores of aerogel, so that the cooperation of CO2 adsorption and catalysis is realized. The three-dimensional aerogel structure is beneficial to high dispersion of active sites and material forming, and has a good industrial application prospect.

The invention belongs to the technical field of synthetic biology and fermentation engineering, and discloses an engineering bacterium for producing Indigoidine based on membrane vesicle engineering reinforced corynebacterium glutamicum and application of the engineering bacterium. According to the invention, the ncp1 gene of corynebacterium glutamicum is knocked out through a CRISPR-Cas12a gene editing technology, connection between cell walls and cell membranes is relieved, and a chassis strain of high-yield extracellular membrane vesicles (OMVs) is constructed; meanwhile, a recombinant expression vector is introduced, indigo synthetase BpsA is positioned and displayed on membrane vesicles by utilizing PorB anchoring protein, and activating enzyme Sfp is co-expressed. In fermentation production, a staged feeding and IPTG (isopropyl-beta-d-thiogalactoside) and Tween-80 dual induction strategy is adopted to promote thalli to express zymoprotein and release a large number of membrane vesicles at the same time. According to the method, the membrane vesicles are used as an extracellular microreactor, so that the'simultaneous synthesis and secretion 'of the indissolvable indigo is realized, the problems of cytotoxicity and metabolic inhibition caused by intracellular precipitation of the product are effectively solved, and the yield and extraction efficiency of the indigo are remarkably improved.

The invention provides an expression vector, an expression system and a preparation method of a recombinant larosidase protein, and belongs to the technical field of recombinant protein preparation, the recombinant expression vector of the larosidase protein provided by the invention comprises an initial vector and a codon-optimized human-derived larosidase coding gene; the nucleotide sequence of the human-derived larosidase coding gene is as shown in SEQ ID NO. 1; according to the recombinant expression system, Chinese hamster ovary cells are taken as host cells, and the recombinant expression vector is transfected. According to the invention, a gene sequence more suitable for expression of Chinese hamster ovary cells is obtained through codon optimization, and then transposon-mediated stable integration is carried out; high-density suspension culture and downstream purification processes are utilized to realize efficient and stable expression of the recombinant larosidase in CHO cells, and the recombinant larosidase with high yield, high activity and high purity can be obtained.

Provided herein are kits, compositions, systems, and methods for treating a subject with a tumor with kynurenine responsive and degrading bacteria (e.g., such that the bacteria reduces or eliminates the tumor). In certain embodiments, the kynurenine responsive bacteria comprise one or more nucleic acid sequences encoding: i) a kynurenine transcriptional regulator protein, ii) a kynurenine transporter protein, iii) a kynureninase (KynU) protein, and iv) at least one protein important or essential for growth of the bacteria expression of which is linked to a kynurenine responsive promoter (e.g., where the corresponding gene(s) has been deactivated or knocked out in the genome of the bacteria).

Methods for determining the function and / or activity and / or potency of isohydrolase proteins are provided.

Disclosed herein are recombinant adeno-associated viral vectors expressing aspartoacylase (ASPA) protein and related uses for treating leukodystrophies.

The present description relates to a recombinant microorganism having production activity of pentamethylenediamine (PMDA), which comprises an E. coli Nissle-derived decarboxylase enzyme protein, and uses thereof.

The present application relates to the field of feed additives, and discloses a lean-type additive for improving the meat quality of Ningxiang pigs and a preparation method thereof, wherein the added enzyme protein carrier has plant activity and enzyme activity, can improve the intestinal health of Ningxiang pigs, promote bone and muscle synthesis, and has antibacterial and antioxidant properties; the bacteria powder microcapsules can realize the rapid colonization of several intestinal probiotics; the bioavailability of plant polyphenols is high; and the amino acid and organic acid compound can promote protein synthesis; the several raw materials used in the present application are green and safe, can have a synergistic effect, can effectively improve the body immunity and the nutrient digestion and absorption capacity of Ningxiang pigs, and can further enhance the constitution of Ningxiang pigs and improve the lean meat rate, while reducing the fecal pollution of Ningxiang pigs and saving the feed cost.

This invention relates to the field of medicinal plant genetic engineering, specifically to a geraniol synthase derived from Morinda officinalis. MoGES Genes and their encoded products and applications. The Morinda officinalis leaf extract enzyme. MoGES The coding region nucleotide sequence of the gene is shown in SEQ ID NO. 1. The Morinda officinalis leaf extract synthase... MoGES The amino acid sequence is shown in SEQ ID NO. 2. It also includes the application of the gene encoding the *Morinda officinalis* geraniol synthase in increasing the geraniol content of plants. This invention successfully cloned the gene encoding *Morinda officinalis* geraniol synthase and verified its efficacy through in vivo functional experiments with *Morinda officinalis*. MoGES To regulate the accumulation of geraniol in Morinda officinalis, an engineered strain BL21(DE3)-MoGES was constructed. A method for the large-scale induction expression and purification of the Morinda officinalis geraniol synthase MoGES protein was developed, and the function of the MoGES protein in catalyzing the synthesis of geraniol in vitro was verified.

The present application relates to a kind of high-efficiency environmental protection crab shell fat removal methods, the crab shell fat removal method includes the following steps: crab shell is dried and crushed, then with water, lipase, protease is mixed, enzymolysis treatment is carried out at 40-50 ℃, and enzyme hydrolysis slurry is obtained;Enzyme hydrolysis slurry is treated with ultrasound at 35-45 ℃, and after treatment mixed slurry is obtained;Mixed slurry is centrifuged, and solid residue is taken, washed and dried, and the crab shell powder after fat removal is obtained.The present application adopts the double coupling process of composite enzymolysis and low-temperature ultrasound, realizes the efficient decomposition and free of crab shell fat, and effective component is kept intact;While any organic solvent and strong acid and alkali are not used in whole process, meet green environmental protection requirement, process is simple, energy consumption is low, can realize continuous industrial production, and production cost is greatly reduced.The method provides a new train of thought for the efficient reuse of crab shell waste, and has economic benefit and environmental benefit.

The invention provides a biosynthesis method of 6-hydroxy-L-tryptophan, on the basis that a novel tryptophan 6-site hydroxylase protein family enzyme is obtained through screening, a cell factory is constructed for biotransformation, and green synthesis of 6-hydroxy-L-tryptophan is achieved. The technical problems of limited types of biocatalysts, insufficient chemical synthesis region selectivity and the like in the prior art are effectively solved. The amino acid sequence of the tryptophan 6-site hydroxylase is shown as SEQ ID NO: 1. The invention provides a protein sequence for catalyzing a new family of tryptophan 6-site hydroxylase, and 6-hydroxy-L-tryptophan can be directly and efficiently synthesized through a cell factory. The method has the advantages of mild reaction conditions, environmental friendliness, high regioselectivity, good product purity, flexible process route and the like, and a brand new technical scheme is provided for industrial production of 6-hydroxy-L-tryptophan.

The invention relates to the technical field of bioengineering and structural biology, and provides a recombinant synechocystis pyruvate dehydrogenase E1 protein crystal which is prepared from a recombinant protein His6-SyPDCsE1, the space group of the crystal is H3, the cell parameters are as follows: a = 146.3, b = 146.3, c = 165.3, and the diffraction resolution is 1.8. The invention also provides a preparation method and application of the crystal. According to the invention, the recombinant protein His6-SyPDCsE1 with high purity and high uniformity is successfully prepared by adopting a double-gene co-expression strategy, and further E1 protein crystal and substrate binding state co-crystal are obtained; the crystal structure fills up the research blank of the prokaryotic synechocystis E1 structure, provides a key basis for revealing a prokaryotic PDHc catalytic mechanism and analyzing the specific structure and function relationship of species, and provides important support for screening drug targets of metabolism-related diseases and agricultural breeding research.

The invention belongs to the technical field of antibiotic enzymes, and relates to a broad-spectrum antibiotic enzyme based on cation amphiphilic (KL) n oligopeptide modification and application thereof. The invention provides a broad-spectrum antibiotic enzyme, the structure of the broad-spectrum antibiotic enzyme comprises an antibiotic enzyme protein ECD7 and an analogue thereof, and a cationic amphiphilic oligopeptide (KL) n modified at the N end or the C end of the antibiotic enzyme protein ECD7 and the analogue thereof, and n in the cationic amphiphilic oligopeptide (KL) n is 5-8. The antibiotic enzyme provided by the invention has a good antibacterial effect on most of common pathogenic bacterial strains, and particularly has stronger antibacterial activity on gram-negative bacteria; in addition, the salt tolerance and fat solubility of the broad-spectrum antibiotic enzyme with the tail end modified with the cationic amphiphilic oligopeptide are remarkably improved, and the broad-spectrum antibiotic enzyme can adapt to wider and more complex production and practical application environments.

The invention provides a preparation method of mannan oligosaccharide, and belongs to the field of biological activity preparation. The preparation method comprises the following steps: fenugreek pretreatment, immobilized enzyme preparation, enzymolysis extraction and extraction. The yield of the mannan oligosaccharide prepared by the method is 40.23 to 41.05 percent, the yield is high, the purity of the crude mannan oligosaccharide is 88.51 to 90.12 percent, the purity is high, the purity of the extracted mannan oligosaccharide is 95.41 to 97.51 percent, and the purity is high; after the immobilized enzyme is oscillated for 24 hours at the speed of 80 r / min in a water tank, the residual enzyme activity is 83.6-85.7%, and the firmness degree of combination of the enzyme protein of the immobilized enzyme and the carrier is high; after 1g of the immobilized enzyme is stored in an environment of 5 DEG C for 240 hours, the residual enzyme activity is 79.9-81.3%, and the storage performance of the immobilized enzyme is strong; after the immobilized enzyme is recycled for six times, the enzyme activity is 68.7-70.5%, the utilization rate of the enzyme is improved, the loading capacity of the carrier is 5.7-6.1%, and the loading capacity is high.

In one aspect, the disclosure relates to a system for site-specific modification of a double­stranded target DNA sequence, the system including at least: a fusion protein comprising an RNA-binding nickase protein and a DNA-dependent DNA polymerase, and a chimeric guide nucleic acid sequence comprising a guide RNA sequence that interacts with the RNA-binding nickase protein and a single-stranded DNA template, wherein the single-stranded DNA template comprises a modified sequence for insertion into a second strand of the double-stranded target DNA sequence and a primer binding site. Also disclosed is a method for site-specific modification of a double-stranded target DNA sequence in a cell. In another aspect, the present disclosure is directed to a fusion protein including an RNA-binding nickase protein and a DNA-dependent DNA polymerase, wherein one or both of the nicakse protein and the DNA polymerase can include one or more mutations to optimize function and prevent off-target effects.

The invention belongs to the technical field of biological catalysis, and particularly relates to an anchor peptide mediated fusion protein and an immobilized enzyme biofilm reactor, the fusion protein comprises a functional enzyme protein and an anchor peptide mutant; the anchoring peptide mutant is obtained by mutating proline at the fifth position of an amino acid sequence as shown in SEQ ID No.2 into valine; the functional enzyme protein is nicotinamide ribokinase or polyphosphate kinase. The functional enzyme protein and the anchoring peptide mutant are connected by using the connecting peptide, and then the obtained fusion protein is anchored on the polypropylene hollow fiber membrane wire material, so that the obtained fusion protein has good anchoring effect and catalytic activity, the batch reaction yield stability can be obviously improved, and the preparation method is suitable for industrial production. An immobilized enzyme bio-membrane reactor is designed, and nicotinamide mononucleotide can be synthesized through biological catalysis.

This invention discloses nanoparticles based on transcytosis-activating ligands, their preparation method, and applications, particularly in the treatment of fundus diseases. The nanoparticles comprise an enzyme protein core and a polymer shell grown in situ on the protein surface. The polymer shell contains positively charged monomers, transcytosis-activating ligands, and neutral monomers. The nanoparticles can enhance the hydrolytic resistance of enzyme proteins (e.g., antioxidant enzymes), protecting them from degradation during transport and improving delivery stability. Simultaneously, the nanoparticles can also promote the efficient delivery of proteases (especially to the posterior segment of the eye), exhibiting good safety and showing excellent application prospects in the medical field.

This invention discloses a BhrPETase keratinase mutant and its application in the degradation of polyethylene terephthalate (PET), belonging to the fields of environmental science and enzyme engineering. The site numbers are based on the amino acid sequence shown in SEQ ID NO:1. The mutant contains substitutions of S14G, H78Y, and S184G relative to SEQ ID NO:1, with the preferred amino acid sequence shown in SEQ ID NO:9. Under conditions of 1 mg enzyme protein / g substrate, 65°C, and 96 h, the preferred mutant exhibits a degradation rate of 90.4% for PET membranes and a half-life of 150 h at 60°C, making it suitable for enzymatic degradation and depolymerization of PET.

The present disclosure provides novel mirror image CRISPR compositions that include components such as mirror image CRISPR-associated endonucleases and mirror image guide RNA for processing of mirror image nucleic acids. In some embodiments, the composition is a mirror image DNA-cleaving and / or modifying composition including a D-form DNA endonuclease (e.g., a D-form Cas protein) and a L-form guide RNA that includes a sequence complementary to a target sequence of a target L-DNA. In some embodiments, the composition further comprises a D-form DNA ligase (e.g., a D-form T4 ligase protein). Also provided are D-form Cas protein and D-form T4 ligase protein compositions, synthetic precursors thereof, and methods of preparing the same.

The application provides a biosynthetic pathway prediction method and system based on deep learning, comprising: obtaining reaction group data; constructing a fingerprint library from the reaction group data; inputting a SMILES sequence of product molecular data into a Transformer model for training; inputting a product molecule to be predicted into the trained Transformer model for precursor molecule prediction to obtain a predicted precursor molecule; forming a complete reaction from the product molecule to be predicted and any predicted precursor molecule and generating a corresponding predicted reaction fingerprint; matching and comparing the predicted reaction fingerprint with the fingerprint library; adding the predicted precursor molecule and enzyme protein corresponding to the reaction with the highest similarity to a synthesis route; determining whether the predicted precursor molecule output by the current Transformer model is a preset building block or whether a preset iteration number is reached; if yes, outputting the current synthesis route; otherwise, inputting the current predicted precursor molecule as a product molecule to be predicted into the Transformer model for precursor molecule prediction.

The present application relates to a kind of natural (6S) -5-methyl tetrahydrofolate and its preparation method, wherein the natural (6S) -5-methyl tetrahydrofolate is prepared by reducing process with folic acid as raw material, and no heavy metal such as platinum or lead, and formaldehyde, benzene sulfonic acid and its ester and other raw materials are used in production process;The content of the natural (6S) -5-methyl tetrahydrofolate is not less than 98%, the amount of JK12A is not higher than 0.1%, methyl tetrahydropterin is not detected, benzene sulfonic acid and its ester are not detected, and enzyme protein residue is not higher than 1 ng / g;The present application has the advantages of convenient post-processing, green environmental protection, high product purity and the like due to the use of green biological production process.