36 results about "Holoenzymes" patented technology

Bis-quinazoline compounds based on the compound (3,4-Dihydro-quinazolin-2-yl)-quinazolin-2-yl-amine, and methods of use of the compounds as inhibitors of bacterial DNA polymerase holoenzymes and in the treatment of bacterial infections are described.

The invention discloses a fungal alpha-amylase-containing beer complex enzyme and a preparation method thereof, belonging to the field of enzyme preparation processing. The high-activity fungal alpha-amylase and other food grade enzyme preparations, plant extracts, antioxidants, protective agents, activating agents and the like can be scientifically compounded by taking concentrated maltase and concentrated malt juice powder as main raw materials; the prepared beer complex enzyme is complete in proteases, high in enzyme activity, difficult to deactivate, mellow in malt aroma, and capable of providing abundant nitrogen source for malt juice, wherein the activity of fungal alpha-amylase in fermenting liquor during the preparation of fungal alpha-amylase is 17000-21000 U / mL. The complex enzyme can be stored for 12 months under the conditions of 0DEG C and 40DEG C, the single enzyme activity loss in the complex enzyme are respectively 0.50% and 0.72%, the enzyme deactivation caused by environment change, and inappropriate operation methods during packaging, storage, transportation, use and the like can be effectively prevented, especially the enzyme deactivation caused by high temperature can be prevented.

The invention discloses a Trichoderma viride F4 strain, and applications thereof. The Trichoderma viride F4 strain is stored at China General Microbiological Culture Collection Center on 10th November 2008, and preservation number is CGMCC No.2736. When the bacterial strain is used for producing cellulase taking pretreated lignocellulose as a carbon source and an inducer, the bacterial strain is capable of tolerating a plurality of inhibitors brought by pretreated lignocellulose, and producing enzyme with high efficiency; the secreted cellulase is comprehensive in composition, enzyme activity is high, and enzymolysis adaptability on pretreated lignocellulose substrate is high.

The invention discloses a beer compound enzyme containing acid protease and a preparation method of the beer compound enzyme containing the acid protease and belongs to the field of enzyme preparation processing. The beer compound enzyme complete in protease system, high in enzyme activity, less prone to inactivation, strong in malt fragrance and capable of providing a rich nitrogen source for wort is mainly prepared from a concentrated malt enzyme and concentrated wort powder by scientifically compounding food-grade enzyme preparations such as the high-activity acid protease, plant extracts, antioxidants, protective agents, activating agents and the like, wherein the enzyme activity of the acid protease in fermentation liquor in an fermentation liquor preparation process is 18000-20000U / mL. When the beer compound enzyme is stored at the temperatures of 0 DEG C and 40 DEG C for 12 months, enzyme inactivation rates of single enzyme in the beer compound enzyme are 0.53% and 0.8% respectively, so that enzyme inactivation caused by environment changes and improper operation methods during packaging, storage, transportation, use and the like can be prevented effectively, and especially, enzyme inactivation caused by high temperature can be prevented.

The invention discloses compound enzyme preparation and the preparing method and application. It includes the following steps: using Aspergillus oryzae as culturing strain; mixing the rice flour or condiment producing main material and fresh bean dregs or bran as the 1:0.5-1 weight ratio; the water content of the mixture is 45%-50%; inoculating 3 per mille-5 per mile the Aspergillus oryzae to culture at 32-35 degree centigrade for 24+-3h; drying under 45 degree centigrade. The produced compound enzyme preparation has full enzyme series, moderate enzyme collocation, proper enzymes, and low price which can be used in fermenting condiment producing technology to obviously improve product quality, shorten production cycle, and reduce production cost.

The invention relates to compositions comprising mixtures of polypeptides having reverse transcriptase (RT) activity and to methods of producing, amplifying or sequencing nucleic acid molecules using these compositions or polypeptides, particularly at temperatures above about 55° C. The invention also relates to nucleic acid molecules produced by these methods, to vectors and host cells comprising these nucleic acid molecules, and use of such nucleic acid molecules to produce desired polypeptide. The invention also relates to methods for producing Avian Sarcoma-Leukosis Virus (ASLV) RT subunits, in particular, Avian Myeloblastosis Virus (AMV) RTs, to isolated nucleic acid molecules encoding ASLV RT subunits, and to ASLV RT subunits produced by these methods. The invention further relates to nucleic acid molecules encoding recombinant RT holoenzymes, particularly ASLV RTs, methods for producing these RTs and to RTs produced by these methods. The invention also relates to kits comprising the compositions, polypeptides, and ASLV RTs of the invention.

The present invention provides a preparation method for high water-holding capacity and water-insolubility corn dietary fiber. The method is a multi-enzyme grading enzymolysis method, and comprises that: adopting processes of neutral protease enzymolysis, high temperature resistance-alpha amylase enzymolysis and glucoamylase combination enzyme enzymolysis; sequentially removing starch, greases, proteins and other impurities from corn bran to obtain pure water-insoluble corn fiber; adopting cellulase to carry out enzymolysis and modification to increase the water-holding capacity of the corn fiber to obtain the high water-holding capacity and water-insolubility corn dietary fiber, wherein the water-holding capacity of the high water-holding capacity and water-insolubility corn dietary fiber is changed into 8.83 g / gDS from the water-holding capacity of the corn bran of 3.44 g / gDS, the improvement range is 157%. The full- enzymolysis method adopted in the invention has characteristics ofrapidness, high performance and no chemical contamination, the equipment is simple, and the method is the potential modified method for the corn dietary fiber.

The invention provides a method for producing recombinant human butyrylcholine esterase on large scale by utilizing a biological platform of the mammary gland of a transgenic animal. The method comprises the following steps: utilizing the biological platform of the mammary gland of the transgenic animal to efficiently produce holonomic and monomeric recombinant human butyrylcholine esterase on large scale for the first time, namely cut-off type enzyme without containing the last 40 amino acid residues of the holonomic butyrylcholine esterase, and a protein fused with monomeric butyrylcholine esterase and albumin. The produced recombinant protein can be used for preventing and treating apnea caused by poisoning of nerve gas, poisoning of organophosphorus pesticide, poisoning of cocaine and succinylcholine, and is used for detecting and removing residues of organophosphorus pesticide on vegetables, melons, fruits, other crops, surfaces of various articles and soil.

The invention relates to a truncated algin lyase gene Aly7B-CDII. The nucleotide sequence of the truncated algin lyase gene Aly7B-CDII is as shown in SEQID NO.1. The algin lyase gene Aly7B-CDII is cloned and transformed to an escherichia coli expression carrier, an escherichia coli recombinant strain capable of performing heterologous expression is obtained, and the strain is used for performing heterologous expression on the truncated body. The enzymology property characterization result indicates that the truncated body Aly7B-CDII exhibits activity on algin, poly M and poly G. The truncatedbody Aly7B-CDII exhibits substrate specificity similar to that of holoenzyme, but the pH and the temperature stability of the truncated body are better than those of the holoenzyme. The truncated algin lyase gene Aly7B-CDII has enormous reference value on reconstructing and exploring of novel algin lyase, and the obtained algin lyase truncated body can be used as a potential tool for producing andpreparing alginate-derived oligosaccharide.

Provided herein are novel compounds that inhibit ribonucleotide reductase (RR) by binding to RRM2 and interfering with the activity of the RRM1 / RRM2 holoenzyme, as well as methods of synthesizing these novel compounds. The compounds may be used to inhibit RR activity and to treat various conditions associated with RRM2 expression, such as for example certain cancer types, mitochondrial diseases, or degenerative diseases.

Atomic coordinates for human serine / threonine protein phosphotase 2A (PP2A) holoenzyme, as well as methods for using these atomic coordinates to prepare inhibitors of PP2A and inhibitors prepared using such methods are provided herein. A biochemical analysis of the interactions of PP2A holoenzyme is also provided. Compositions including mimetics and small molecules of the invention and, optionally, secondary agents may be used to treat disorders in which PP2A activity plays a contributing role.

The present invention is generally related to compositions and methods for the reverse transcription of nucleic acid molecules, especially messenger RNA molecules. Specifically, the invention relates to compositions comprising mixtures of polypeptides having reverse transcriptase (RT) activity, and to methods of producing, amplifying or sequencing nucleic acid molecules (particularly cDNA molecules) using these compositions or polypeptides, particularly at temperatures above about 55° C. The invention also relates to nucleic acid molecules produced by these methods, to vectors and host cells comprising these nucleic acid molecules, and to the use of such nucleic acid molecules to produce desired polypeptides. The invention also relates to methods for producing Rous Sarcoma Virus (RSV) and Avian Myeloblastosis Virus (AMV) RTs or other Avian Sarcoma-Leukosis Virus (ASLV) RTs (α and / or β subunits thereof), to isolated nucleic acid molecules encoding such RSV RT, AMV RT or other ASLV RT subunits, to vectors and host cells comprising these isolated nucleic acid molecules and to RSV RT, AMV RT and other ASLV RT subunits produced by these methods. The invention further relates to nucleic acid molecules encoding recombinant heterodimeric RT holoenzymes, particularly heterodimeric RSV RTs, AMV RTs or other ASLV RTs (which may be αβ RTs, ββ RTs, or α RTs), vectors (particularly baculovirus vectors) and host cells (particularly insect and yeast cells) comprising these nucleic acid molecules, methods for producing these heterodimeric RTs and heterodimeric RTs produced by these methods. The invention also relates to kits comprising the compositions, polypeptides, or RSV RTs, AMV RTs or other ASLV RTs of the invention.

The invention provides a method to identify inhibitors of the formation of holoenzyme from core RNA polymerase and sigma.

A process for biologically synthesizing the extracellular cholesterol oxidase from Brevibacterium is disclosed. The high-output bacterium DGCDC-82 is fermented in optimized condition to obtain said enzyme up to 700-1000 U / L. After purified, its specific enzyme activity can reach 30 U / mg. Said enzyme contains coenzyme FAD and cysteine in the active center of enzyme. In the dual-phase (water phase and organic phase) system, said enzyme can be used to uniquely oxidize the cholesterol to becone cholest-4-ene-3-ketone with 92% of transfer rate, which can be used to reduce cholesterol, blood fat and low-density lipoprotein, so having wide application range.

The invention discloses a preparation method of a high-activity composite enzyme for feed, belonging to the technical field of preparation of an enzyme preparation for feed. An Aspergillus niger DM-18 strain for a high-yield composite enzyme for feed, which is used as an initial strain, is subjected to liquid submerged mixed fermentation in an optimized culture medium under specific fermentation conditions by an improve fermentation technique to obtain the composite enzyme for feed, which has the advantages of complete enzymatic system, high enzyme activity, high temperature resistance and wide pH value resistance range. The prepared crude enzyme fermentation liquid contains multiple enzyme activities, wherein the proteinase activity is 6500-6700 U / ml, the mannase activity is 1500-1700 U / ml, the alpha-galactase activity is 1200-1300 U / ml, and the pectinase activity is 1000-1200 U / ml. The enzymes still have higher enzyme activity (up to 80% or above) at 30-75 DEG C under the pH value of 2-7; and thus, the composite enzyme is more suitable for the demands of a processing technique in feed industry, and the processed feed has low loss of enzyme activity.

The invention discloses a flora for degrading corn straw (hereinafter referred to as straw) to produce polysaccharide, and belongs to the technical field of solid waste disposal. The mixed flora comprises streptomyces griseorubens, streptomyces carpaticus and streptomyces rochei, and the inoculation amount is 1:1:1. The degrading strains are respectively named ye-9, er-72 and se-93, and are preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation numbers are respectively 18289, 18290 and 18291, and the preservation date is July 24th, 2019. According to the invention, a single strain is primarily screened from a sample, and the primarily screened single strain is secondarily screened by taking the activities of four enzymes, namely cellulose whole enzyme, incision enzyme, excision enzyme and beta-glucosidase, as indexes. The strains which are obtained by secondary screening and have a good degradation effect are obtained through screening, the enzyme activity is determined, and efficient mixed flora is picked out. The straw degradation effect of the mixed flora is better than that of a single strain, and the polysaccharide yield is as high as 2159.23 mg / L under the optimized fermentation condition. A basis is provided for microbial degradation of the straws, and resource utilization of the straws is realized.

A preparation method for a feeding solid complex enzyme comprises the following contents of: a. drying solid fermented materials by low temperature flash evaporation, and crashing the materials to prepare the raw powder; b. adding auxiliary ingredients to the raw powder according to the prescription to prepare the feeding solid complex enzyme; and c. packaging and storing the finished product. Inthe prescription, the auxiliary ingredients include acid protease, one or two types of alpha-amylases, and corn bran powder. The ratio of the components by weight is as follows: 4-12 percent of raw powder, 0 or 1-6 percent of 50000U / g acid protease, 0 or 4-5 percent of 2000U / g alpha-amylase, and the balance of corn bran powder. The solid complex enzyme prepared by the method has the advantages oflow cost, complete zymogram and high enzyme activity. Furthermore, the enzymes are mutually supplementary and dependent without antagonism.

The invention discloses a preparation method of a high-activity composite enzyme for feed, belonging to the technical field of preparation of an enzyme preparation for feed. An Aspergillus niger DM-18 strain for a high-yield composite enzyme for feed, which is used as an initial strain, is subjected to liquid submerged mixed fermentation in an optimized culture medium under specific fermentation conditions by an improve fermentation technique to obtain the composite enzyme for feed, which has the advantages of complete enzymatic system, high enzyme activity, high temperature resistance and wide pH value resistance range. The prepared crude enzyme fermentation liquid contains multiple enzyme activities, wherein the proteinase activity is 6500-6700 U / ml, the mannase activity is 1500-1700 U / ml, the alpha-galactase activity is 1200-1300 U / ml, and the pectinase activity is 1000-1200 U / ml. The enzymes still have higher enzyme activity (up to 80% or above) at 30-75 DEG C under the pH value of 2-7; and thus, the composite enzyme is more suitable for the demands of a processing technique in feed industry, and the processed feed has low loss of enzyme activity.

The invention discloses a fungal alpha-amylase-containing beer complex enzyme and a preparation method thereof, belonging to the field of enzyme preparation processing. The high-activity fungal alpha-amylase and other food grade enzyme preparations, plant extracts, antioxidants, protective agents, activating agents and the like can be scientifically compounded by taking concentrated maltase and concentrated malt juice powder as main raw materials; the prepared beer complex enzyme is complete in proteases, high in enzyme activity, difficult to deactivate, mellow in malt aroma, and capable of providing abundant nitrogen source for malt juice, wherein the activity of fungal alpha-amylase in fermenting liquor during the preparation of fungal alpha-amylase is 17000-21000 U / mL. The complex enzyme can be stored for 12 months under the conditions of 0DEG C and 40DEG C, the single enzyme activity loss in the complex enzyme are respectively 0.50% and 0.72%, the enzyme deactivation caused by environment change, and inappropriate operation methods during packaging, storage, transportation, use and the like can be effectively prevented, especially the enzyme deactivation caused by high temperature can be prevented.

The invention belongs to the technical field of pharmaceutical analysis, and discloses an experimental method for screening a fatty acid synthase (FASN) inhibitor in a peony seed meal monomeric compound based on computer simulation, and the experimental method for screening the FASN inhibitor in the peony seed meal monomeric compound based on computer simulation comprises the following specific operation steps: S1, molecular docking; s2, cell activity detection; s3, performing protein immunoblotting; and S4, cell apoptosis detection. Whether four monomeric compounds in the peony seed meal extract have anti-tumor activity caused by inhibition of FASN activity and expression or not is screened by means of a molecular docking method, a homologous modeling means is tried to construct a whole enzyme dimer structure of FASN, and a new thought and a new method are provided for screening of the FASN inhibitor. A large number of invalid experiments can be effectively avoided, the success rate of experiment operation is improved, and the experiment operation cost is reduced.

The invention discloses novel lysyl endopeptidase. An enzyme core peptide is connected with histidine tags through an appropriate flexible linker peptide, so that nanogram residual enzyme in a samplecan be detected by an ELISA (enzyme-linked immunosorbent assay) method; and total activity of wild lysyl endopeptidase is ensured. At the same time, the invention further provides a preparation methodof the lysyl endopeptidase. Compared with a gene engineering modification and fermentation technology, the preparation method has the benefit of high yield; a recovery amount of 857mg novel lysyl endopeptidase can be obtained after purification of 1L fermentation liquid; and the method has an industrial potential.

The invention discloses compound enzyme preparation and the preparing method and application. It includes the following steps: using Aspergillus oryzae as culturing strain; mixing the rice flour or condiment producing main material and fresh bean dregs or bran as the 1:0.5-1 weight ratio; the water content of the mixture is 45%-50%; inoculating 3 per mille-5 per mile the Aspergillus oryzae to culture at 32-35 degree centigrade for 24+-3h; drying under 45 degree centigrade. The produced compound enzyme preparation has full enzyme series, moderate enzyme collocation, proper enzymes, and low price which can be used in fermenting condiment producing technology to obviously improve product quality, shorten production cycle, and reduce production cost.

The invention relates to the technical field of polypeptide production, and in particular relates to a production process of full-enzyme polypeptide for a fertilizer. The production process comprisesthe steps of 1) current-assisted swelling: putting protein into a swelling solution, introducing constant-voltage forward current for treatment, and introducing constant-voltage reverse current for treatment; (2) ultrasonic-assisted enzymolysis: adding an activated compound enzyme preparation into a product obtained in the step (1) for enzymolysis, meanwhile, carrying out intermittent ultrasonic treatment in the enzymolysis process, and circulating the operation; and (3) enzyme deactivation: carrying out enzyme deactivation on the product obtained in the step (2), and then carrying out freezedrying to obtain the full-enzyme polypeptide for the fertilizer. The full-enzyme polypeptide for the fertilizer has better physiological activity, permeability and stability, is easily absorbed by crops, is not easily inactivated by environmental influence, has low chelating process conditions, and can chelate metal ions with different chemical activities. The polypeptide disclosed by the invention further has the capability of inhibiting harmful bacteria in soil, and is wide in antibacterial spectrum.

A method for replicating and amplifying a target nucleic acid sequence is described. A method of the invention involves the formation of a recombination intermediate without the prior denaturing of a nucleic acid duplex through the use of a recombination factor. The recombination intermediate is treated with a high fidelity polymerase to permit the replication and amplification of the target nucleic acid sequence. In preferred embodiments, the polymerase comprises a polymerase holoenzyme. In further preferred embodiments, the recombination factor is bacteriophage T4 UvsX protein or homologs from other species, and the polymerase holoenzyme comprises a polymerase enzyme, a clamp protein and a clamp loader protein, derived from viral, bacteriophage, prokaryotic, archaebacterial, or eukaryotic systems.

The invention provides a preparation method of water-insoluble corn dietary fiber with a high oil-holding capacity. The preparation method adopts a multi-enzyme successive enzymatic hydrolysis technology, wherein the multi-enzyme successive enzymatic hydrolysis technology is an enzymolysis technology adopting a neutral protease and a high-temperature resistant alpha-amylase and glucoamylase composite enzyme. The preparation method comprises the following steps of removing orderly impurities of starch, proteins and the like in corn husks to obtain purified water-insoluble corn fiber, and carrying out enzymolysis modification of the water-insoluble corn fiber by xylanase to obtain the water-insoluble corn dietary fiber with a high oil-holding capacity, wherein the oil-holding capacity is improved 81.7% from 2.02g / gDS to 3.67g / gDS and a oil-holding capacity of corn husks is 2.02g / gDS. The preparation method adopts a full enzymolysis technology. Through the preparation method, starch is removed; a water-insoluble fiber component in a product is retained; and subsequent neutralization treatment processes are avoided. In addition, the preparation method is utilized for processing and utilization of waste corn husks from starch and alcohol industries, and can reduce environmental pollution and improve a degree of comprehensive development and utilization of a by-product.

The invention discloses alginate lyase AlyPL17 and a truncation thereof. The invention further clones and transforms genes of the AlyPL17 and the truncation thereof onto an escherichia coli expression vector to obtain an escherichia coli recombinant strain capable of heterologous expression, and the strain is used for heterologous expression of corresponding alginate lyase. Results of enzymatic property characterization show that the optimum temperature of the AlyPL17 and the truncation thereof is 45 DEG C, the optimum pH is 9.0, and the AlyPL17 has high specific activity. From the aspect of action mode, the whole enzyme AlyPL17 of the enzyme adopts a mixed action mode, and truncation bodies at two ends adopt an incision action mode, which indicates that the unique action mode of the enzyme is caused by the synergistic effect of the truncation bodies at the two ends. The invention has great reference value for analyzing the alginate lyase in a mixed action mode, and the obtained alginate lyase AlyPL17 can be used as a potential tool for converting a brown algae resource into a biological energy source.

The present disclosure describes purified telomerase holoenzyme and its delivery to cells, such as T cells, for increasing telomere length, increasing cell proliferation, and impeding cell senescence.