52results about How to "Broad substrate specificity" patented technology

The invention discloses a method for preparing micromolecular collagen peptide from fish scales by utilizing complex enzyme. The method comprises the steps of deashing and degreasing fish scales used as raw materials; adding complex enzyme mixed by pepsin, papain and trypsin for hydrolyzing, inactivating the enzyme, carrying out coarse filtering, film filtering, vacuum concentrating and freeze drying to obtain the micromolecular collagen peptide. The collagen peptide has the average molecular weight of 500-1,000Da, can be absorbed and utilized by human bodies easily, can be applied to cosmetic, healthcare products, foods, beverages and other industries, and has remarkable economic effect and social effect.

The present invention provides a method of detecting an enzyme in a sample wherein the enzyme is capable of adding or removing a chemical moiety to or from a nucleic acid molecule, which thereby confers altered sensitivity of the nucleic acid molecule in a subsequent process. The invention also relates to diagnostic methods which take advantage of the method of the invention and kits which are useful for carrying out the method of the invention.

The present invention is related to a fungal serine protease enzyme, which comprises an amino acid sequence the mature Fa_RF7182 enzyme having an amino acid sequence of SEQ ID NO: 18. The serine protease is obtainable from Fusarium acuminatum, more preferably from the deposited strain CBS 124084. Also disclosed are nucleic acid sequences encoding said protease, such as plasmid pALK2530 comprising the nucleotide sequence SEQ ID NO:12 deposited in Escherichia coli RF7803 under accession number DSM 22208 and plasmid pALK2531 comprising the full-length gene SEQ ID NO: 13 deposited in E. coli RF7879 under accession number DSM 22209, as well as fungal hosts, such as Trichoderm. Said protease is useful as an enzyme preparation applicable in detergent compositions and for treating fibers, for treating wool, for treating hair, for treating leather, for treating food or feed, or for any applications involving modification, degradation or removal of proteinaceous material.

The invention provides a novel phthalate hydrolase gene derived from a soil metagenome library. The nucleotide sequence and an amino acid sequence of the novel phthalate hydrolase gene are shown as SEQ ID NO.1 and SEQ ID NO.2. After the esterase gene is connected to an expression vector pET28a (+), the esterase gene is transformed into escherichia coli BL21 (DE3) to realize heterologous expression. The molecular weight of the purified recombinase (EstJ6) is 33.31 kDa. The EstJ6 has wide substrate specificity on phthalate, and the EstJ6 not only can hydrolyze phthalate with a simple side chain,but also can hydrolyze diethyl hexyl phthalate and monoethyl hexyl phthalate with complex and longer side chains. In addition, site-specific mutagenesis experiments show that the catalytic triad residue of EstJ6 is S146-E240-H270, and mutation of any amino acid in the three causes the EstJ6 to lose the catalytic ability. The novel phthalate hydrolase disclosed by the invention can be applied to the fields of food industry, agriculture, biotechnology and the like due to the specific activity and enzymatic characteristics of the novel phthalate hydrolase.

The present invention provides fusion polypeptides comprising polypeptide ligands that are modified by circular permutation and fused to at least one polypeptide fusion partner wherein such fusion polypeptides have new, improved or enhanced biological functions or activities. Such improvements include, but are not limited to, increased binding affinity, increased activity, increased agonist activity (super agonist), antagonist activity, increased accessibility, increased flexibility of the active site, increased stability, broader and / or changed substrate specificity, and combinations thereof.

The invention relates to a non-bitter fishy smell, instant whole egg powder and a preparation method thereof. The method comprises: (a) adopting two-stage dynamic high-pressure micro-jet treatment; (b) sequentially adding compound protease and phospholipid to the whole egg liquid Enzyme A1; wherein the composite protease is papain and flavor protease, the dosage ratio of the two is 2:1, the amount of composite protease added in the whole egg liquid is whole egg liquid: composite protease=100:0.8-1.2, add The amount is calculated by mass ratio; phospholipase A1 is enzymatically hydrolyzed at room temperature, and the added amount in the whole egg liquid is whole egg liquid: phospholipase A1=100:0.01‑0.05, and the added amount is calculated by kg / L; (c) After spray drying, whole egg powder is fluidized bed granulated with a binder. The invention cooperates with dynamic high-pressure micro-jet homogenization, enzymatic hydrolysis and fluidized bed technology to obtain whole egg powder with good instant solubility, and no stratification is found within 10 hours after brewing.

The invention provides a neutral high temperature xylanase CtXyn10A as well as genes and application thereof. The neutral high temperature xylanase CtXyn10A has proteins of amino acid sequences shown in SEQ ID No. 3 and / or SEQ ID No. 5 in a sequence table; and / or is characterized in that amino acid residue sequences shown in SEQ ID No. 3 and / or SEQ ID No. 5 in the sequence table are replaced and / or deleted and / or added by one or several amino acid residues, and have proteins with xylanase activity and derived from the proteins with the amino acid sequences shown in SEQ ID No. 3 and / or SEQ ID No. 5 in the sequence table. The xylanase CtXyn10A has an optimum pH of 6.0 to 7.0 and an optimum temperature of 70 DEG C, keeps enzyme activity of 20% or above at 90 DEG C and pH 9.0, and specific activity is 461 U / mg; the xylanase CtXyn10A has the activity of xylanase, glucanase and cellulase, is easy to produce by industrial fermentation, and can be widely used in food, paper, energy industry and the like as a novel broad-spectrum enzyme preparation.

The invention discloses an alginate lyase SHA-I gene with a nucleotide sequence shown in SEQ ID NO: 1. The invention constructs a prokaryotic expression vector of the alginate lyase gene (i) SHA-I, and the prokaryotic expression vector is capable of acquiring an expression product alginate lyase SHA-I in a relatively short time, and has broad substrate specificity that PolyM and PolyG can both be used as a substrate. The alginate lyase has enzyme activity reaching 13U / mg and is a bifunctional enzyme with broad prospect. The prokaryotic expression vector and the overall expression system provided by the invention are easy to operate, and convenient for industrial production.

The present invention relates to microbacterium, broad spectrum glycosaminoglycan lyase expressed by the microbacterium, and a coding gene and applications of the broad spectrum glycosaminoglycan lyase. According to the present invention, the Microbacterium sp. WS15 strain is preserved in the China General Microbiological Culture Collection Center (CGMCC) on December 05, 2016, and has the preservation number of CGMCC No.13421, wherein the address is Institute of Microbiology, Chinese Academy of Sciences, 3# Court No.1, Beichen West Road, Chaoyang District, Beijing; and the glycosaminoglycan lyase TT16 is firstly obtained from the genome of Microbacterium WS15, can degrade the hyaluronic acid with no sulfation modification, can further degrade chondroitin sulfate A (CS-A), chondroitin sulfate C (CS-C) and chondroitin sulfate E (CS-E) being subjected to different sulfation modifications, and is the broad spectrum glycosaminoglycan lyase.

The present invention relates to the bioupgrading of crude oil is directed to a process for decreasing the acidity of an acidic crude oil, comprising contacting an acidic crude oil with a mixture nitrogen containing compounds selected from the group comprising ammonia, ammonia hydroxide, amines and the salts thereof, and in the presence of lipase enzyme, under conditions of suitable temperature and pressure sufficient to form the corresponding amide. The resulting naphthenic acid derived amides can then be processed normally in a refinery using such processes as cracking or hydrotreating and converted to hydrocarbon, ammonia and carbon dioxide without causing damage to the refinery infrastructure. This enzyme process is done at reduced temperatures (40-60° C.) and pressures requiring less energy.

The present invention provides a novel glucuronosyltransferase, a polynucleotide encoding the same (e.g., a polynucleotide comprising a polynucleotide consisting of the nucleotide sequence at positions 1 to 1362 in the nucleotide sequence represented by SEQ ID NO: 7, or a polynucleotide comprising a polynucleotide encoding a protein having the amino acid sequence represented by SEQ ID NO: 8); and so on. A novel glucuronosyltransferase having a broad substrate specificity and others can thus be provided.

The present disclosure is relates to novel proteins and peptides having novel and / or enhanced functions and / or behaviors with respect to a native protein or peptide, and methods of making the novel proteins and peptides using techniques of circular permutation and protein engineering.

The invention discloses a prokaryotic expression vector pGEX-4T-1-ZHO-I for efficiently expressing sphingomonassp.ZHO alginate lyase ZHO-I. The vector is a prokaryotic expression vector containing sphingomonassp.ZHO alginate lyase gene ZHO-I. The prokaryotic expression vector disclosed by the invention can obtain an expression product alginate lyase ZHO-I in a short time, and the obtained recombinant alginate lyase ZHO-I has wide substrate specificity; both PloyG and PloyM can be used as a substrate, and the enzyme activity can reach 53.2U / mg; the prokaryotic expression vector is a difunctional enzyme with broad application prospect; and the prokaryotic expression vector and the whole expression system disclosed by the invention are easy to operate and is favorable for the industrial production.

The invention relates to the field of gene engineering, in particular to neutral low-temperature xylosidase CaXyl43A and a gene and application thereof. The amino acid sequence of the xylosidase is shown in SEQ ID NO.1. The xylosidase has the properties including the optimal pH of 6.8, the optimal temperature of 35 DEG C, the enzyme activity maintenance property at 0 DEG C, and the specific activity of 181.6 U / mg, has the activities of xylosidase, Arabinfuranosidease and circumscribed xylanase, and has certain xylose tolerance and the capacity of completely degrading xylo-oligosaccharide to generate xylose. Industrial fermentation production is easy. As a novel broad-spectrum enzyme preparation, the xylosidase can be widely used for aquatic feed industry, food industry, papermaking industry, energy industry and the like.

The invention discloses an alginate lyase SHA-3 gene with a nucleotide sequence as shown in SEQ ID NO:1. The invention establishes a prokaryotic expression vector of the alginate lyase SHA-3 gene. The vector can obtain an expression product, namely alginate lyase SHA-3 within relatively-short time, has wide substrate specificity, can take polymannuronic acid PolyM or polyguluronic acid PolyG as a substrate, and has the enzyme activity of 12U / mg so as to be a difunctional enzyme with a wide application prospect. The prokaryotic expression vector and a whole expression system are easily operated and are conveniently used for realizing industrial production.

The invention discloses an alginate lyase SHA-2 gene. A nucleotide sequence is as shown in SEQ ID NO:1. A prokaryotic expression vector of the alginate lyase SHA-2 gene is constructed; an expression product alginate lyase SHA-2 can be obtained by the prokaryotic expression vector within a relatively short period of time, and has wide substrate specificity; poly-mannuronate (PolyM) and poly-guluronate (PolyG) can also be utilized as substrates; the enzyme activity can reach 5.2U / mg; the alginate lyase SHA-2 gene is a bifunctional enzyme with a wide application prospect; the prokaryotic expression vector and an overall expression system are easy to operate; and industrialized production is facilitated.

The invention relates to prokaryotic expression and application of lipase and an immobilization method, and belongs to the fields of enzyme engineering and molecular biology application. A lipase Tm1350 gene is derived from an extreme thermophilic bacterium. The method comprises the following steps: constructing the gene in a prokaryotic expression carrier pET-28a, obtaining a recombinant plasmid pET-28a-Tm1350, transferring into a host cell E.coli BL21, and inducing expression of the recombinant lipase; purifying and identifying the enzymatic property; and constructing a shuttle expression vector pHS-CotB-Tm1350 with a high copy number, transferring into a bacillus subtilis strain DB403, inducing generation of recombinant spores, and assembling along with capsid protein CotB, wherein the enzyme is co-expressed on the spore surfaces. According to the immobilization method, lipase is immobilized through a spore surface technology for the first time; the lipase has good heat stability, alkali resistance, methanol activity and methanol resistance, and has great industrialized application potential, for example, preparation of biodiesel. The enzyme is shown on the spore surfaces, is relatively good in stability, can be recycled by simple centrifugation or filtration, and has a good application prospect.

The invention relates to a method for extracting panax quinquefolius polysaccharides from panax quinquefolius fibrous roots. The method utilizes the panax quinquefolius fibrous roots as raw materials for extraction and comprises sequential steps as follows: grinding, alkaline leaching, enzymolysis with compound enzyme, extraction with an ultrasonic method, concentration, ethanol precipitation and vacuum drying. According to the method, a traditional water or ethanol extraction method for the panax quinquefolius polysaccharides is broken through, an alkaline solution with certain concentration is utilized to promote the physicochemical reaction in panax quinquefolius polysaccharide grains or among the panax quinquefolius polysaccharide grains for alkaline leaching extraction, the extraction is assisted with composite enzymatic hydrolysis and ultrasonic wave technologies through combination of a wall-breaking effect of cellulase on plant cells and a specific degradation effect of protease, the process is simple and easy to operate, the extraction rate and the yield of the panax quinquefolius polysaccharides are increased remarkably, and theoretical reference bases are provided for industrial production of the extraction of the panax quinquefolius polysaccharides.

The invention discloses an alginate lyase SHA-6 gene with a nucleotide sequence as shown in SEQ ID NO:1; the alginate lyase SHA-6 gene is recombined with pET-32a(+) plasmids for being converted into escherichia coli BL21 to obtain engineering bacteria of alginate lyase SHA-6; the bacterial strain is induced to express to generate a great number of inclusion body proteins SHA-6; the alginate lyaseSHA-6 inclusion body is subjected to protein purifying and renaturation, so that the inclusion body recovers activity; the re-natured alginate lyase has wide substrate specificity, polymannose aldehyde acid PolyM and polygulose aldehyde acid PolyG can be utilized as a substrate, and enzyme activity reaches 13.5U / mg, so that the alginate lyase SHA-6 gene is alginate lyase with a wide application prospect.

Strict anaerobic thermophilic bacterium belonging to the group of Thermoanaerobacter mathranii and mutants and derivatives thereof. The bacterium is particularly suitable for the production of fermentation products such as ethanol, lactic acid, acetic acid and hydrogen from lignocellulosic biomass.

The invention discloses a thermophilic esterase derived from an Aquifex aeolicus strain and an expression thereof and belongs to the technical field of biomedical engineering. A novel thermophilic esterase gene is found, three expression systems are established, and efficient expression and enzymatic property research are achieved. A eukaryotic expression system is established, wherein a carrier pPIC9K is preferred for expression vector establishment, a pichia pastoris host is converted, GS115 is preferred, and efficient expression is achieved. A prokaryotic escherichia coli expression system is established, wherein a carrier MBP3 is preferred for expression vector establishment, an escherichia coli host is converted, BL21 and Origami2 are preferred, and efficient expression is achieved. A prokaryotic bacillus megatherium expression system is established, wherein a carrier pHIS1525 is preferred for expression vector establishment, a bacillus megatherium host is converted, YYBm1 is preferred, and efficient expression is achieved. The recombinant enzyme has the advantages of esterase activity, thermophily, thermal stability and the like and has great potential in industrial applications under the high temperature condition.

There is provided an acetogenic microbial cell which is capable of producing at least one higher alcohol from a carbon source, wherein the acetogenic microbial cell is genetically modified to comprise an increased expression relative to its wild type cell of at least one enzyme, E8, a butyryl-CoA:acetate CoA transferase (cat3). There is also provided a method and use of the cell to produce higher alcohols.

A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) adding a precipitation buffer to a sample to prepare an acidic precipitation mixture wherein said precipitation buffer comprises a metal cation precipitant and a buffering agent, has a pH value of 4.0 or less and does not comprise an organic solvent selected from aprotic polar solvents and protic solvents and wherein the acidic precipitation mixture comprises the metal cation precipitant in a concentration of less than 200 mM and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. The present method allows to avoid the use of organic solvents during protein precipitation. Also provided is a precipitation buffer.

A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) preparing a precipitation mixture by adding at least one metal cation precipitant and at least one organic solvent selected from aprotic polar solvents and protic solvents to the sample, wherein the precipitation mixture i) comprises the metal cation precipitant; ii) comprises the organic solvent in a concentration of 15% or less; iii) comprises a buffering agent; and iv) has an acidic pH value, and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. Using an organic solvent as claimed during the protein precipitation step in the defined concentration provides a supernatant which in addition to small RNA also comprises large RNA. This is an advantage as the present method provides more flexibility to the user. The described method can be used for isolating and hence analysing different target RNAs.

A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) preparing a precipitation mixture by adding at least one metal cation precipitant and at least one organic solvent selected from aprotic polar solvents and protic solvents to the sample, wherein the precipitation mixture i) comprises the metal cation precipitant; ii) comprises the organic solvent in a concentration of 15% or less; iii) comprises a buffering agent; and iv) has an acidic pH value, and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. Using an organic solvent as claimed during the protein precipitation step in the defined concentration provides a supernatant which in addition to small RNA also comprises large RNA. This is an advantage as the present method provides more flexibility to the user. The described method can be used for isolating and hence analysing different target RNAs.

The invention discloses a sphingomonas alginate lyase gene ZH0-II and an efficient prokaryotic expression vector pGEX-4T-1-ZH0-II thereof. A Ptac promoter is utilized to control the expression of the prokaryotic expression vector pGEX-4T-1-ZH0-II in colon bacillus. The expression of the alginate lyase gene is performed by the colon bacillus expression vector, so that the alginate lyase ZH0-II as an express product can be obtained in a short time. The obtained recombinant alginate lyase gene ZH0-III is wide in substrate specificity, wherein the PloyG or the PloyM can be served as a substrate; and the enzyme activity can reach 61.7U / mg. The prokaryotic expression vector and the overall expression system are easy to operate, and are convenient for industrialized production.

The invention relates to the field of genetic engineering, in particular to a neutral low-temperature xylanase CaXyn10A, and a gene and application thereof. The amino acid sequence of the neutral low-temperature xylanase CaXyn10A is shown as SEQ ID NO. 1. The xylanase is characterized in that the optimal pH is 6.0-6.5, the optimal temperature is 40 DEG C, over 20% of enzyme activity is kept at the temperature of 0 DEG C, and the specific activity is 453U / mg; the xylanase has activities of xylanase, glucanase and cellulose; industrialized fermentation production is facilitated. The neutral low-temperature xylanase CaXyn10A serving as a novel broad-spectrum preparation can be widely applied to aquatic feed, food, papermaking, energy industry and the like.

The present technology pertains methods for converting hydrolyzed lignocellulosic biomass material to high levels of a biofuel using extreme theromophilic bacterial strains.

The invention discloses an alginate lyase SHA-6 gene with a nucleotide sequence as shown in SEQ ID NO:1; the alginate lyase SHA-6 gene is recombined with pET-32a(+) plasmids for being converted into escherichia coli BL21 to obtain engineering bacteria of alginate lyase SHA-6; the bacterial strain is induced to express to generate a great number of inclusion body proteins SHA-6; the alginate lyaseSHA-6 inclusion body is subjected to protein purifying and renaturation, so that the inclusion body recovers activity; the re-natured alginate lyase has wide substrate specificity, polymannose aldehyde acid PolyM and polygulose aldehyde acid PolyG can be utilized as a substrate, and enzyme activity reaches 13.5U / mg, so that the alginate lyase SHA-6 gene is alginate lyase with a wide application prospect.