328 results about "Asparagine" patented technology

The invention discloses a formate dehydrogenase mutant and application thereof in catalytic synthesis of D-mannitol, and belongs to the technical field of bioengineering. According to the formate dehydrogenase mutant provided by the invention, the 18th-site glutamic acid of wild-type formate dehydrogenase with an amino acid sequence shown as SEQ ID NO.2 is mutated into proline, the 57th-site asparagine is mutated into glutamic acid, the 70th-site histidine is mutated into tryptophan, the 235th-site serine is mutated into threonine, and the 316th-site valine is mutated into threonine, so that the formate dehydrogenase mutant is high in enzyme activity and catalytic activity; the coenzyme circulation efficiency during the production of D-mannitol can be obviously improved, so that the yield of D-mannitol can be effectively improved. When the genetically engineered bacterium co-expressed by the formate dehydrogenase mutant and the mannitol dehydrogenase mutant is used for catalyzing 150 g / L fructose to produce D-mannitol, the yield of D-mannitol within 15 h can reach 140 g / L or above.

The invention discloses a high-performance carbonyl reductase mutant and application thereof in synthesis of a series of chiral alcohols. The high-performance carbonyl reductase mutant is obtained by performing the following mutations on an amino acid sequence as shown in SEQ ID NO.2: the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine, and the 69th alanine mutates into any one of lysine, leucine, aspartic acid or asparagine; and / or the 100th glycine is mutated into any one of glutamic acid, serine, lysine or asparagine. Compared with a wild type enzyme, the thermal stability and the catalytic activity of the obtained mutant are remarkably improved, particularly, single point mutants Mut-A69N, Mut-G100E and Mut-G100N and a combined mutant Mut-A69N-G100N show excellent relative enzyme activity, thermal stability and substrate tolerance. The mutant has high catalytic activity and stability to various chiral alcohols, and the contradiction that the traditional carbonyl reductase is easy to inactivate at high temperature and the catalytic efficiency is difficult to achieve at the same time is solved. The invention provides an efficient and stable enzyme catalysis tool for green biological preparation of chiral drug intermediates.

The present application relates to a pyridoxal kinase mutant, a recombinant expression vector and a microbial cell and their applications, and belongs to the technical field of biological catalysis. In order to solve the problem of low product concentration in existing enzyme catalysis, a pyridoxal kinase mutant is provided, the amino acid sequence is selected from the amino acid sequence shown in SEQ ID NO. 1, the lysine at position 229 is mutated to alanine, phenylalanine, methionine, arginine, threonine, histidine, serine, tyrosine, valine, leucine, isoleucine, proline, asparagine, aspartic acid or glutamic acid; the pyridoxal kinase mutant is used for catalyzing pyridoxal to synthesize pyridoxal phosphate, and a recombinant expression vector and a microbial cell can be further formed. The present application has good enzyme activity, high product conversion rate, high concentration of pyridoxal phosphate obtained, and the concentration of the product catalyzed by the wild-type pyridoxal kinase to phosphorylate pyridoxal is obviously improved.

In the first aspect, the present disclosure provides a Lactoferrin polypeptide fragment, where the Lactoferrin polypeptide fragment has an amino acid sequence shown in SEQ ID NO: 1. In the second aspect, the present disclosure provides a preparation method of the Lactoferrin polypeptide fragment, including: protecting amino acids using a 9-fluorenylmethoxycarbonyl (Fmoc) protecting group, coupling the amino acids with a P-hydroxymethylphenoxymethyl polyethylene resin (HMP resin) in an order of cysteine, alanine, leucine, cysteine, glutamate, threonine, asparagine, aspartate, asparagine, phenylalanine, leucine, leucine, asparagine, and lysine, and separating the HMP resin to obtain a target Lactoferrin polypeptide fragment. In the third aspect, the present disclosure further provides an anti-Lactoferrin antibody prepared using the Lactoferrin polypeptide fragment, and use of the Lactoferrin polypeptide fragment in preparation of an anti-Lactoferrin flow cytometric antibody and in preparation of a Lactoferrin detection product.

The invention is suitable for the technical field of soybean breeding, and provides a KASP molecular marker for soybean plant height identification and application of the KASP molecular marker. The KASP molecular marker is based on an SNP variation site of a Glyma. 19G206100 gene between two parents of Chinese flat stem soybean and F02, the mutation position of the Glyma. 19G206100 gene is the 312th basic group on a CDS sequence, C of a reference genome is mutated into G, the 104th amino acid on a protein sequence is mutated into lysine from asparagine, and the 104th amino acid on the protein sequence is mutated into lysine from asparagine. The primer of the KASP molecular marker comprises a forward primer with a nucleotide sequence as shown in SEQ ID NO.1-2 and a reverse primer with a nucleotide sequence as shown in SEQ ID NO.3. The invention further discloses a kit for detecting the KASP molecular marker. The KASP molecular marker is developed on the basis of the polymorphic site of Gm19G206100 in germplasm resources, the marker has the advantages of being high in resolution ratio and easy to detect, can be applied to plant height selection of early generations of soybean cross breeding, has the effects of reducing the breeding scale, reducing the workload and improving the breeding efficiency, and has important application value in production.

According to the scheme, the invention provides the recombinant macrolide enzyme as well as the preparation method and the application thereof, the amino acid sequence of the recombinant macrolide enzyme is as shown in SEQ ID NO.1, and the recombinant macrolide enzyme is derived from an EreC esterase family of enterobacter hormaechei and is obtained through site-specific mutagenesis; the mutation sites are as follows: glutamic acid at the 44th site is mutated into asparagine, tyrosine at the 55th site is mutated into proline, proline at the 76th site is mutated into arginine, phenylalanine at the 153rd site is mutated into alanine, and serine at the 219th site is mutated into glutamic acid. The method is expected to remove macrolide antibiotic pollution, and has good economic benefits and practical values.

The invention relates to a chicken beta-defensin-6 mutant based on AI aided design, a recombinant expression system and application, and relates to the technical field of gene engineering and protein engineering. The chicken beta defensin-6 mutant has the following amino acids at the following five sites: lysine at the 33rd site, lysine at the 35th site, asparagine at the 49th site, arginine at the 61st site and proline at the 65th site. The chicken beta-defensin-6 mutant has high stability, activity and solubility, compared with wild chicken beta-defensin-6, the chicken beta-defensin-6 mutant has higher antibacterial activity and expression level, an efficient and safe antibacterial substitute is provided for livestock and poultry breeding, and the chicken beta-defensin-6 mutant can be widely applied to the fields of livestock and poultry breeding, animal health care and biological preservation.

In order to improve aggregation property of a PPR protein, the A6 amino acid of the 1st PPR motif from the N-terminus (M1) is made more hydrophilic. Further, the A9 amino acid of M1 is made to be a hydrophilic amino acid or glycine. The A6 amino acid is preferably asparagine or aspartic acid, and the A9 amino acid is preferably glutamine, glutamic acid, lysine, or glycine. Proteins containing such a PPR motif as M1 motif may have not only improved aggregation property, but also high binding power to a target nucleic acid.

The invention discloses a fermentation process for shortening a fermentation period and efficiently synthesizing Ectoine. The fermentation process comprises the following steps: screening and activating high-yield strains, preparing a fermentation culture medium, performing fermentation culture, performing sectional dissolved oxygen regulation and control, performing double-stage pH dynamic regulation and control, performing carbon-nitrogen ratio dynamic regulation based on metabolic flow analysis, performing osmotic pressure gradient induction in the later stage of fermentation, and extracting and purifying a product. According to the fermentation process for shortening the fermentation period and efficiently synthesizing the Ectoin, the expression quantity of key enzymes for synthesizing the Ectoin is increased by 80% or above compared with that of an original strain through the Halomonassp.ECT-09 strain bred through ultraviolet-nitrosoguanidine compound mutation, the basic yield reaches 35 g / L, a foundation is laid for high yield, and the method is suitable for industrial production. A glucose-trehalose mixed carbon source and a yeast extract-ammonium nitrate mixed nitrogen source are adopted, and an L-aspartic acid-L-asparagine mixed precursor is added, so that the metabolic pathway is shortened, and the loss of an intermediate product is reduced.

The present invention relates to an isolated peptide comprising the amino acid sequence of Formula (I): (O)n-K-(O)m-Z; wherein K is lysine optionally modified with a self-immolative moiety Z is tryptophan or is absent; O is GHGX1Y (SEQ ID NO: 1); G is glycine, H is histidine, Y is tyrosine; each X1 is independently selected from alanine, glycine, serine, asparagine, histidine (H), arginine (R), aspartic acid, tyrosine, and proline; n is 0 – 5; m is 0 – 5; n+m is 3, 4, 5, 6, 7 or 8, and optionally, wherein not all of the X1 residues are proline (P). Such peptide is a coacervate forming peptide. The present invention also relates to methods of preparing coacervate compositions comprising the isolated peptide, optionally also comprising one or more biomacromolecules (payloads). The present invention further relates to the coacervate compositions per se, and their uses, e.g. in methods of treatment or diagnosis.

A CVB virus expression vector comprising a PIV5 W3A viral genome comprising a mutation at amino acid residue S157 or S156 of the P / V gene and a deletion of the small hydrophobic (SH) gene of the PIV5 W3A viral genome, wherein the amino acid substitution at amino acid residue S157 or S156 comprises a substitution of serine (S) with phenylalanine (F) or asparagine (N), and the SH gene has a deletion of the SH open reading frame or the entire SH gene transcription unit. The CVB virus expression vector expresses a heterologous polypeptide, including SARS-CoV-2 spike (S), and / or nucleocapsid (N) and / or membrane (M) proteins, RSV fusion protein (F), or other antigens.

An Fc-binding polypeptide with improved alkaline stability comprising a mutant of the Fc-binding domain of Staphylococcus protein A (SpA), said mutant being defined by SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 22, SEQ ID NO 51 or EQ ID NO 52, wherein at least the asparagine or serine residue at a position corresponding to position 11 in SEQ ID NO: 4-7 has been mutated to an amino acid selected from the group consisting of glutamic acid, lysine, tyrosine, threonine, phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, alanine, histidine and arginine.

This invention belongs to the field of compound synthesis, specifically relating to a method for synthesizing D-2,3-diaminopropionic acid. The method includes first reacting D-Boc asparagine and sodium hypohalate in a solvent to obtain an intermediate solution; then adjusting the pH of the intermediate solution to 4-8, removing the solvent to obtain a solid; finally, reacting the solid with acid and purifying it to obtain the final product; wherein the solvent in step (1) is a sodium hydroxide solution. This invention ensures a rapid and complete reaction by controlling the temperature and time of the reaction process; and by adding concentrated hydrochloric acid in stages, it can obtain the deprotected target product in high yield. The method of this invention avoids the use of expensive iodine reagents, uses inexpensive and readily available raw materials, has a high overall reaction yield, low cost, and is suitable for industrial production.

The application provides an arginine mutant and application thereof in 1,4-butanediamine biosynthesis, and belongs to the field of bioengineering. Escherichia coli The mutant is obtained by site-directed mutagenesis of key site amino acid residues based on wild-type EcAdiA from strain The application improves the enzyme activity stability of arginine decarboxylase in neutral environment, improves the conversion rate and conversion speed of 1,4-butanediamine in neutral environment, and lays a foundation for high-efficiency biosynthesis of 1,4-butanediamine.

The invention discloses a phytase mutant AppA as well as a coding gene, a preparation method and application thereof. The phytase mutant AppA is obtained by mutating amino acids at the 137th site, the 185th site and the 255th site of wild type phytase AppA from asparagine, aspartic acid and tyrosine into valine, asparagine and aspartic acid respectively; the amino acid sequence and the nucleotide sequence of the gene are SEQ ID NO.1 and SEQ ID NO.2 respectively. The catalytic efficiency of the mutant enzyme AppA provided by the invention on sodium phytate is improved by 2.5 times compared with that of a wild enzyme AppA; under the condition of 70 DEG C, the half-life period of the mutant enzyme AppA is 3.4 times that of the wild enzyme AppA. The mutant enzyme AppA has the characteristic of high enzyme activity under a high-temperature condition, which indicates that the mutant enzyme AppA has an important application prospect in industrial production of a feed in which phytic acid needs to be degraded under the high-temperature condition.

The application discloses a beta-1,4-xylanase mutant and a construction method thereof, and comprises the following steps: constructing a beta-1,4-xylanase recombinant expression vector; introducing the recombinant vector pNZ8048-P5-XynA into a competent lactococcus lactis by using an electrotransformation technology to construct a wild-type expression vector; on the basis, designing and constructing a multi-site mutant recombinant plasmid by using a site-directed mutation method, wherein the eighth asparagine (Asn) of the beta-1,4-xylanase is mutated into tyrosine (Tyr), the 22nd serine (Ser) is mutated into proline (Pro), and the 59th alanine (Ala) is mutated into lysine (Lys); introducing the multi-site mutant recombinant plasmid into the competent lactococcus lactis to construct a heterologous expression strain of the mutant beta-1,4-xylanase (Mut-1); and stably obtaining the mutant xylanase beta-1,4-xylanase (Mut-1) with high activity by using a metal affinity chromatography (Ni 2+ ) method. The specific enzyme activity of the mutant reaches 1929.30±5.36 U / mg, and the optimal temperature is increased by 15 DEG C compared with that of the wild type.

This application relates to the field of tumor treatment technology, specifically to a drug combination and evaluation method for improving the sensitivity of MSS CRC to anti-PD-1 / PD-L1 therapy. The drug combination comprises vancomycin and an immune checkpoint inhibitor. Vancomycin is used to remodel the gut microbiota and reduce L-asparagine levels; the immune checkpoint inhibitor is used to activate CD8. + T-cell anti-tumor response. By combining vancomycin with immune checkpoint inhibitors, the antigen-presenting capacity of dendritic cells is enhanced, thereby increasing the sensitivity of MSS CRC to anti-PD-1 or anti-PD-L1 immunotherapy. This application focuses on the upstream initiation link of gut microbiota-metabolites-dendritic cells-immune activation, relieving the inhibition of dendritic cell antigen presentation by L-asparagine and enhancing CD8+. + T cell activation and tumor immune response enhance the sensitivity of MSS-type colorectal cancer to anti-PD-1 / PD-L1 therapy.

The invention discloses a generalized pustular psoriasis diagnosis marker based on metabonomics and application of the generalized pustular psoriasis diagnosis marker. The diagnostic marker is prepared from one or more of the following 35 compounds: pyruvic acid, alpha-ketoisovaleric acid, 2-hydroxybutyric acid, 3-hydroxybutyric acid, methane thiophosphoric acid, proline, uracil, tranexamic acid, 4-aminobutyric acid, threonine, scopoletin, dodecanol, N-methyl-L-leucine, L-cysteine-glycine and L-kynurenine. The feed additive is prepared from the following raw materials: 3-hydroxybenzoic acid, allantoin, delta-tocopherol, xylofuranose, glucose-1-phosphoric acid, pyrophosphate, taurine, L-asparagine, phthalic acid, 4-(dimethylamino) azobenzene, 5-tert-butyl-1h-indole-2, 3-dione, quinic acid, glucose, histidine, lysine, palmitic acid, 7-methylguanine, oleic acid and whale acid. The marker can be used for accurately distinguishing patients with generalized pustular psoriasis from healthy people.

To provide use methods of compositions in drug production for inhibiting cancer cells having low asparagine synthetase (ASNS) expression.SOLUTION: The disclosure relates to a use method of a composition comprising an arginine reducing compound and an asparagine reducing compound for preparing a medicament to inhibit a cancer cell, where an arginase being the arginine reducing compound is specifically effective in combination use with an asparaginase being the arginine reducing compound for inhibiting proliferation of cancer cells, in particular, cells having low ASNS expression. Further, the medicament comprises a compound that reduces glutamine concentration such as an aminotransferase inhibitor.SELECTED DRAWING: Figure 1

The present invention relates to an antibody or binding fragment thereof that binds to a protein, preferably a T cell receptor, comprising an epitope comprising or consisting of an amino acid sequence having at least 80 %, such as 83 %, preferably at least 90 %, for instance 100 % sequence identity to the amino acid sequence set forth in SEQ ID NO: 62 (EVPKXiR), wherein Xi is selected from Alanine (A), Arginine (R), Asparagine (N), Aspartic acid (D), Cysteine (C), Glutamine (Q), Glutamic acid (E), Glycine (G), Histidine (H), Isoleucine (I), Leucine (L), Lysine (K), Methionine (M), Phenylalanine (F), Proline (P), Serine (S), Threonine (T), Tryptophan (W), Tyrosine (Y), Valine (V). Further, the invention relates to pharmaceutical compositions comprising such an antibody or binding fragment thereof and to the use of such an antibody or binding fragment thereof for use as a medicament or for use as a screening agent.

This invention provides an induction culture medium and method for using marigold anthers as explants. The induction culture medium is based on MS solid medium containing agar, with specific amounts of sucrose, tryptophan, phenylalanine, arginine, casein amino acids, anhydrous asparagine, 6-benzylpurine, naphthaleneacetic acid, silver nitrate, pyridoxine hydrochloride, thiamine hydrochloride, nicotinic acid, D-biotin, folic acid, ascorbic acid, and coconut juice added. This invention also provides a method for induction culture using this induction culture medium. This invention can greatly improve the callus induction rate of marigolds, significantly increasing the number of regenerated seedlings, and provides technical guidance for the cultivation of haploid marigolds using anthers.

The invention relates to the technical field of crop planting, in particular to application of aspartic acid and asparagine in plant planting. The initiator comprises an aspartic acid solution and / or an asparagine solution; the concentration of the aspartic acid solution is 25 [mu] M; the concentration of the asparagine solution is 1000 [mu] M; the invention also discloses application of aspartic acid and asparagine in preparation of a seed initiator. According to the application of the aspartic acid and the asparagine in plant planting, the aspartic acid and the asparagine are used as initiators, the problems of existing initiators are solved, meanwhile, the aspartic acid and the asparagine are plant endogenous amino acids and are non-toxic, biodegradable and environmentally friendly, meanwhile, the initiators are easy to obtain, the using method is simple, and the application range is wide. The method is suitable for large-scale agricultural application.

An affinity membrane separator is provided that includes a separation substrate with a plurality of peptide ligands positioned thereon. The peptide ligands preferentially bind to one of a nucleic acid product and a waste nucleic acid product, e.g., produced by a mRNA synthesis bioreactor. The peptide ligands include fewer than 20 residues and at least one defined secondary structure, and have a global charge greater than about 1 and at least one lysine, arginine, or glutamine residue in order to preferentially bind ds-RNA relative to ss-RNA, or a global charge less than about 0 and at least one serine or asparagine residue in order to preferentially bind ss-RNA relative to ds-RNA. Peptide ligands can advantageously bind to ds-RNA byproducts from the production of ss-RNA vaccines, e.g., against SARS-CoV-2, while allowing the ss-RNA vaccines themselves to pass through the separator, providing an efficient system for production of purified ss-RNA vaccine products.

The application provides a polypeptide, a plant immune elicitor and application thereof. The polypeptide has 10 or 11 amino acid residues; the amino acid at the first position of the polypeptide is serine, and the amino acids at the 9th and 10th positions are both cysteine; if the polypeptide has 11 amino acid residues, the amino acid at the 11th position of the polypeptide is asparagine. The polypeptide can solve the problem that there is no small peptide secreted by gramineae plants in the prior art, and is suitable for the field of plant immune elicitors.

The present application relates to a kind of methyltransferase CicOMT10 mutant, preparation method, biological material and application.The methyltransferase mutant is mutated with the amino acid sequence such as SEQ ID NO:1 CicOMT10 enzyme, and the mutation includes: the asparagine of 179th is mutated to alanine;And / or, the threonine of 316th is mutated to alanine.The methyltransferase mutant of the present application can efficiently convert eriodictyol into hesperetin.

The invention provides a novel herbicide-resistant mutation efficiently created by editing a rice genome targeted G-to-T basic group. Specifically, the invention provides a separated herbicide resistant polypeptide, and the herbicide resistant polypeptide is a mutant ALS1 polypeptide, the mutant ALS1 polypeptide is mutated at the 171st amino acid, corresponding to SEQ ID NO.1, of the wild ALS1 polypeptide, and proline (P) at the 171st position is mutated into one or more amino acids selected from the following groups: threonine (T), histidine (H), asparagine (N) and arginine (R). According to the invention, it is found for the first time that a guanine base editor is used for editing the wild ALS1 polypeptide in a plant; efficient G-to-T base editing can be achieved, multiple amino acid replacement types which are not reported before are created, and partial mutation types can greatly improve the tolerance of plant herbicides.

The application discloses a high-temperature-resistant alkaline alpha-galactosidase mutant and a preparation method thereof. The mutant is obtained by performing site-directed mutation on alpha-galactosidase derived from Anoxybacillus vitaminiphilus, and at least one amino acid in three positions of 444th, 457th and 509th in the amino acid sequence shown in SEQ ID No. 1 is replaced. The asparagine Asn in the 444th position is mutated into phenylalanine Phe, the aspartic acid Asp in the 457th position is mutated into methionine Met, and the arginine Arg in the 509th position is mutated into tryptophan Trp. The alpha-galactosidase mutant has good thermal stability and alkali stability by performing site-directed mutation on alpha-galactosidase derived from Anoxybacillus vitaminiphilus (Anoxybacillus vitaminiphilus WMF1).

The present invention relates to a monoclonal antibody reacting with a glycopeptide and use thereof. The present invention provides a monoclonal antibody which is an epitope of both a fucose moiety and an amino acid of a peptide moiety. The antibody of the present invention is a monoclonal antibody reacting with a glycopeptide containing a core fucose and an amino acid of 4 or more residues continuously from an asparagine of an additional sugar chain toward a C-terminal side, the antibody being an epitope of both the core fucose of the glycopeptide and an amino acid of the glycopeptide which is 3 or more residues from the asparagine of the additional sugar chain toward the C-terminal side.

The present disclosure provides mutated PglB oligosaccharyltransferase enzymes that have the ability to efficiently catalyze the transfer of a saccharide from a lipid carrier to an asparagine reissue in a glycosylation motif on a protein. Also provided are polynucleotides encoding the mutated PglB oligosaccharyltransferase enzymes, host cells capable of expressing the engineered PglB oligosaccharyltransferase enzymes, and methods of using the engineered PglB oligosaccharyltransferase enzymes to make N-glycosylated proteins. Also provided are N-glycosylated proteins that are made using the engineered PglB oligosaccharyltransferase enzymes.