405 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 invention discloses a complete medium for culturing DF-1 cells, including DMEM basal medium 13.7g / L, insulin, non-essential amino acids, transferrin, glutathione, ascorbyl phosphate, sodium selenite, ethanolamine, hydrocortisone, and newborn calf serum. Wherein non-essential amino acids include glycine, alanine, proline, serine, and asparagine. A complete medium for culturing DF-1 cells provided by the present invention can well promote the proliferation of DF-1 cells, can achieve the same proliferation effect as high-concentration fetal bovine serum, and is suitable for the adherent culture of DF-1 cells; using low-concentration newborn calf serum, it is possible to greatly avoid the batch differences, unstable culture results, cytotoxicity, and high risk of carrying pathogens brought by the culture medium with high-concentration serum, narrow application range, expensive fetal bovine serum, and high culture cost; the components are simple, the culture medium preparation process is short in time consumption, the culture efficiency is high, the repeatability is good, and it is suitable for batch production.

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.

The invention discloses an Escherichia coli endogenous alcohol dehydrogenase EcYjgB mutant and application thereof. The mutant is obtained by performing any one of the following mutations (a) to (c) on Escherichia coli endogenous alcohol dehydrogenase EcYjgB as shown in SEQ ID NO.1: (a) mutating serine at the 200th site into arginine; (b) the serine at the 199th site is mutated into aspartic acid, and the serine at the 200th site is mutated into arginine; and (c) the serine at the 199th site is mutated into aspartic acid, the serine at the 200th site is mutated into arginine, and the asparagine at the 240th site is mutated into tryptophan. The mutant is used for efficiently catalyzing reduction of HMF or furfural to respectively synthesize BHMF and furfuryl alcohol, so that the purposes of improving the production efficiency, reducing the production cost and improving the environmental effect are achieved.

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 present invention relates generally to glutamine-free cell culture media supplemented with asparagine. The invention further concerns the production of recombinant proteins, such as antibodies, in asparagine-supplemented glutamine-free mammalian cell culture.

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.

Disclosed herein is a delivery agent that facilitates effective delivery of drugs and other compounds across lipid layers. The delivery agent disclosed herein comprises an asparagine-containing mask peptide group comprising at least one asparagine amino acid moiety and which provides lipid solubility under selected conditions and aqueous solubility under other conditions, and can effectively deliver compounds into the cell cytosol.

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.

The invention discloses a carbonyl reductase mutant for synthesizing a chiral alcohol sulfone compound and application of the carbonyl reductase mutant, a mutant M4A1 of carbonyl reductase ChKRED20 is used as a female parent, serine at the 188th site is mutated into any one of asparagine, glycine or threonine, and the carbonyl reductase mutant for synthesizing the chiral alcohol sulfone compound is obtained. Specifically, a mutant M4A1 of ChKRED20 is taken as a female parent, serine at the 188th site is mutated into asparagine, and a mutant ChKRED20-M4C4 is obtained. The mutant M4A1 of ChKRED20 is used as a female parent, serine at the 188th site is mutated into glycine, and the mutant ChKRED20-M4C3 is obtained. The mutant M4A1 of ChKRED20 is used as a female parent, serine at the 188th site is mutated into threonine, and the mutant ChKRED20-M4C6 is obtained. The three mutants can be used as catalysts to synthesize chiral alcohol sulfone compounds, and especially have great advantages in asymmetric synthesis of alpha-substituted beta-alcohol sulfone.

The invention provides a lipid nanoparticle conjugate as well as a preparation method and application thereof. In particular, an antibody is coupled to the surface of the LNP through an N-glycosylation site of an Fc heavy chain by using a copper-free click chemical reaction, thereby obtaining the LNP having a specific cell or tissue targeting property. Preferably, the N-glycosylation site is located on the asparagine residue at the 297th position of the Fc heavy chain.

There is provided peptidic compounds of Formula I, A or B(Rradn6-[linker]-RL-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-ψ-Xaa9-NH2). Xaa1 is D-Phe, Cpa, D-Cpa, Nal, D-Nal, 2-Nal, or D-2-Nal; Xaa2 is Asn, Gln, Hse, Cit or His. Xaa3 is Trp, Bta, Trp(Me), Trp(7-Me), Trp(6-Me), Trp(5-Me), Trp(4-Me), Trp(2-Me), Trp(7-F), Trp(6-F), Trp(5-F), Trp(4-F), Trp(5-OH), or αMe-Trp. Xaa4 is Ala or Ser. Xaa5 is Val, Cpg, or Tle. Xaa6 is Gly, NMe-Gly, or D-Ala. Xaa7 is His or NMe-His. Xaa8 is Leu or Phe. Xaa9-NH2 is a C-terminally amidated amino acid residue selected from Pro, 4-oxa-L-Pro, Me2Thz, or Thz. ψ represents a peptide bond or reduced peptide bond joining Xaa8 to Xaa9. Rradn6 is 1-5 radiolabeling groups. There is also provided the use of such compounds as imaging agents or therapeutic agents.

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 invention relates to a surface display system based on an isopeptide bond and a construction method and application of the surface display system, two conformational proteins including an anchor protein containing the isopeptide bond and a target protein are designed through a modularization method, and through covalent coupling formed between lysine and asparagine or aspartic acid residues in a molecular adhesive system based on the isopeptide bond, the surface display system based on the isopeptide bond is obtained. And displaying the target protein on the surface of escherichia coli. According to the surface display system based on the isopeptide bond, high-efficiency display of complex enzyme and macromolecular protein on the cell surface is realized, and the influence of direct fusion with anchoring protein on the activity of target protein is avoided. According to the constructed surface display whole-cell catalyst, the tedious enzyme purification process is omitted, the stability of the enzyme is improved, the production cost is lower, and the surface display whole-cell catalyst shows a good application prospect in biological catalysis.

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.