34 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 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 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.

The invention relates to high-level production of pHBA in green plants using a unique expression cassette. The latter comprises a chorismate pyruvate lyase (CPL) coding sequence operably linked to a suitable promoter capable of driving protein expression in higher plants. Additionally, the CPL cassette comprises a sequence encoding a chloroplast transit peptide, its natural cleavage site, and a small portion of the transit peptide donor protein fused to the N-terminus of CPL. The chloroplast targeting sequence targets the foreign protein to the chloroplast compartment and aids in its uptake into the organelle. The cleavage site is unique to the transit peptide, and cleavage of the chimeric protein encoded by the cassette at this site releases a novel polypeptide that has full enzyme activity, comprising the mature CPL enzyme and a small portion of the transit peptide donor.

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 an angle dioxygenase gene dpeA1A2 and its application. An angle dioxygenase gene dpeA1A2, which is composed of the angle dioxygenase large subunit encoding gene dpeA1 whose size is 1383bp shown in SEQ ID NO.1, and the angle dioxygenase gene dpeA1 whose size is 561bp shown in SEQ ID NO.2 Oxygenase small subunit coding gene dpeA2 and the connecting sequence GATG between the two components. The DpeA1A2BC holoenzyme provided by the present invention can completely degrade 100 mg / L of diphenyl ether within 72 hr; in addition, the DpeA1A2BC holoenzyme can also completely degrade 100 mg / L of 2-phenoxybenzoic acid (2-PBA) within 72 hr. Therefore, the angle dioxygenase gene dpeA1A2 is used in the construction of transgenic crops for degrading diphenyl ether or 2-phenoxybenzoic acid. DpeA1A2BC holoenzyme is used in the degradation of diphenyl ether and 2‑PBA.

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 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.

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 discloses an alginate lyase AlyPL17 and its truncated body, and further transforms the gene clone of AlyPL17 and its truncated body into an expression vector of Escherichia coli to obtain a recombinant strain of Escherichia coli capable of heterologous expression, using the strain Heterologous expression of the corresponding alginate lyase. The characterization results of enzymatic properties showed that the optimum temperature of AlyPL17 and its truncation was 45℃, the optimum pH was 9.0, and the enzyme had high specific activity. From the mode of action, the holoenzyme AlyPL17 of this enzyme adopts a mixed mode of action, while the two-terminal truncation adopts an endo-interaction mode, indicating that the unique mode of action of this enzyme is caused by the synergistic action of the two-terminal truncation. The invention has great reference value for analyzing the mixed action mode alginase, and the obtained alginate lyase AlyPL17 can be used as a potential tool for transforming brown algae resources into bioenergy.

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 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.