86results about How to "Taller than alive" patented technology

The invention provides a method for preparing heparinase I. The method for preparing the heparinase I comprises the following steps of: inoculating flavobacterium heparinum serving as a raw material to a seed culture medium for culture; then inoculating the flavobacterium heparinum to a fermentation culture medium; centrifugally collecting precipitate; performing ultrasonication on the precipitate; performing centrifugation again to obtain crude enzyme liquid of the flavobacterium heparinum heparinase I; and performing SP-sepharose FF chromatographic purification on the crude enzyme liquid for three times to obtain the high-purity flavobacterium heparinum heparinase I, wherein the SP-sepharose FF chromatographic purification for three times is protected by calcium chloride in the whole course, so that the yield of pure enzymic activity is greatly increased. The method for preparing the heparinase I has the characteristics of simple process, easy amplification, large preparation amount of products at a time, low cost of reagents and the like. The specific activity of the prepared heparinase I reaches 223 IU/mg and the yield of the pure enzymic activity reaches 30 percent. Compared with the conventional newest method, the method for preparing the flavobacterium heparinum heparinase I has the advantages of increasing the specific activity by more than two times, and the purification yield by about one time.

This invention relates to a method for producing recombined human granular leukocyte colony stimulating factors including: fermenting, breaking bactrriums, extracting occlusion bodies, chromatographing with molecular sieves, renaturating, exchanging anions, exchanging cations to get the raw fluid, in which, BL21 is selected as the host bacterium and the engineering bacterium type is got in high expression volume, quick reproduction and stable passage, several purification steps greatly reduce the residural toxin in the bacterium and the specific activity is increased greatly, a dialysis method is applied for the renaturation to reduce the concentration of the denaturalization agent steadily so the renaturation rate is at high level.

The invention discloses a cephalosporin acylase mutant and an encoding gene and an application thereof. The protein provided by the invention is any one of the following 1)-3): 1) protein formed by an amino acid sequence shown as sequence 4 in the sequence table; 2) protein formed by an amino acid sequence shown as sequence 3 in the sequence table; 3) protein which is obtained by substitution and/or deletion and/or addition of one or several amino acid residues in the amino acid residue sequence of sequence 3 or sequence 4 in the sequence table, has the functions of cephalosporin acylase, andis derived from 1). Experiment of the invention shows that the invention provides a wild-type CPC acylase mutant; HPLC results show that the specific activity for CPC of the wild-type CPC acylase mutant of the invention is increased by 6.5 times when compared with that of wild-type enzymes. In one-step conversion, the conversion rate is up to above 98% within 3 hours.

The invention relates to a method for extracting superoxide dismutase, in particular to a method for extracting superoxide dismutase from barbadosnut. The method for extracting superoxide dismutase is achieved by using seeds or leaves of barbadosnut as raw materials, and comprises the steps of the preparing crude extract, removing foreign protein, sectional salting-out ammonium sulfate, precipitating acetone, carrying out Sephadex G2100 molecular sieve column chromatography, and the like. The invention provides a new source for extracting superoxide dismutase, and optimizes technical parameters for extracting superoxide dismutase. The method for extracting superoxide dismutase can give consideration to both the specific activity and the recovery ratio of enzyme, and is beneficial to the large-scale production of enterprises.

The invention discloses lipase made of aspergillus niger strains, and a producing method and utilization thereof. The lipase (Aspergillus niger) AN0512 is preserved in the China general microbiological culture collection center which is called CGMCC for short on November 19th, 2012, and the preserving number is CGMCC No.6835. The producing method includes: fermenting the lipase in a culture medium using olive oil as a main carbon source; removing thalluses to obtain lipase crude enzyme liquid; and then depositing and dialyzing, and separating the lipase crude enzyme liquid through anion exchange chromatography and molecular exclusion chromatography to obtain electrophoresis pure lipase. The lipase has a wide temperature application range, good activity when hydrogen ion concentration ranges from 4.0 to 6.0, and good acid resistance, and can be used in industries of food processing, drug preparing, chemical synthesizing and the like.

The invention relates to a purifying method of interferon. The method comprises the following steps of a. performing fermentation culture by selecting engineering bacteria as bacterial strains; b. separating and washing inclusion bodies of the bacterial strains obtained from the fermentation culture to obtain refined inclusion bodies; c. performing renaturation on the refined inclusion bodies to obtain a renaturation preoduct; d. performing anionic column chromatography treatment on the renaturation product to obtain an anionic column chromatography product; d. performing cation column chromatography treatment on the anionic column chromatography product to obtain a cation column chromatography product; and f. processing the cation column chromatography product by using a reverse phase packing column chromatography, and performing desalinizing treatment to obtain the interferon.

The invention discloses a technology for producing and recombining a human tectotype profibrinolytic activator TNK mutant with Cytopore porous microcarrier culture method, which comprises the following steps: suspending TNK-tPA engineering project CHO cell strain with the culture medium; culturing by transferring to the cell culture bottle; forming dense monolayer cell; culturing by transferring the cell to Wheaton agitator; transferring to the Celligen reactor (the culture temperature is 37 Deg. C, PH is 7.1+-0.1, the dissolved oxygen is 30-60%, the original rotating speed is 20rpm, the rotating speed increases progressively after 4 hours and reaches 40rpm when inoculating for 6 hours); culturing 20 days by replenishing the culture liquid and the fresh microcarrier; collecting when the cell concentration reaches the finite level. TNK-tPA with the invention has the big cell density, the low cost, the high expression level and the high expression product specific activity.

The invention discloses a DNA (deoxyribonucleic acid) sequence for coding a green plant bug water-soluble trehalase. The invention also discloses a green plant bug water-soluble trehalase, a recombinant expression vector of the DNA sequence for coding the green plant bug water-soluble trehalase, a transgenic cell system or transgenic recombinant bacterium, application of the recombinant expression vector and transgenic cell system or transgenic recombinant bacterium in producing the green plant bug water-soluble trehalase, and a preparation method of the green plant bug water-soluble trehalase. The molecular weight of the prepared zymoprotein is 71KD, the optimum pH value is 7.0, the optimum temperature is 55 DEG C, and the zymoprotein can degrade trehalose into glucose. The purified trehalase can be used for qualitative and quantitative work of trehalose content in industry and some other fields, and can also provide foundation for researching and developing insect trehalase inhibitors.

The invention aims at providing bacteria for producing organic solvent resisting lipase, the organic solvent resisting lipase produced from the bacteria and the application of the organic solvent resisting lipase to the enzymatic reaction for the synthesis of the biodiesel in the organic phase, in particular to the ester exchange reaction for the synthesis of the biodiesel. The bacteria of pseudomonas aeruginosa LX1 is obtained through screening, the organic solvent resisting LX1 lipase produced from the pseudomonas aeruginosa LX1 has the amino acid sequence showed in the sequence identifier (SEQ ID) No. 2, and the organic solvent resisting LX1 lipase encoding gene has the nucleotide sequence showed in the SEQ ID No. 2. The organic solvent resisting LX1 lipase is used for catalyzing the substrate of soybean oil and exchanging the soybean oil and the ester methanol in the organic solvent tert-butanol system or the solvent-free system to synthesize the clean energy of biodiesel which has the conversion rate of 80-90 percent.

The invention discloses a preparation method of a human blood coagulation factor VIII and a human blood coagulation factor VIII product and relates to the field of blood products. The preparation method of the human blood coagulation factor VIII comprises the following steps: dissolving a cryoprecipitate with a 0.015-0.025mol/L tromethamine solution in a mass ratio of (3-5): 1 by reasonably processing the cryoprecipitate; and carrying out separation and purification by means of a polyethylene glycol precipitating method combined with ion exchange chromatography, wherein the recovery ratio of the human blood coagulation factor VIII and the specific activity of a final product can be improved effectively, the yield reaches 180-240IU/L plasma, and the specific activity is not lower than 100IU/mg proteins. The prepared human blood coagulation factor VIII product is rich in vWF factors and the proportion of the vWF factors and the human blood coagulation factor VIII is close to 1: 1. Besides treating hemophiliac A, the human blood coagulation factor VIII can be also used for treating patients with angiohemophilia, and the human blood coagulation factor VIII has good stability and heat resistance.

The invention provides a method for removing viruses in preparation of antitoxin and antiserum. The method comprises the following steps of: diluting immunological blood plasma and adjusting pH value; then digesting by stomach enzyme; heating to denaturize; adsorbing by alum; concentrating/desalinating by ultrafiltration; performing ion exchange chromatography to obtain collected liquid; removing bacteria and filtering; and then subpackaging to obtain an antitoxin and antiserum product. Thus, the viruses which probably exist in raw material serum can be removed simultaneously in the process of preparing the antitoxin and antiserum. The virus removal method for the antitoxin and antiserum provided by the invention has the characteristics of simplicity, quickness economy and practicality; the viruses which probably exist in the raw material serum can be removed completely during production to guarantee the quality of the product.

The invention provides a high-specific-activity high-catalytic-efficiency amylase mutant and a preparation method and application thereof and relates to the field of gene engineering. Acid amylase derived from Talaromyces leycettanus JCM12802 is used as a female parent, and the wild type is subjected to site-directed mutagenesis by means of molecular biological techniques. Under the modification condition, the amylase mutant has significantly higher catalytic efficiency and specific activity than the wild (before mutation) female parent. By using the scheme, it is possible to greatly increase the specific activity and acting conditions of amylase, and basis is provided for the amylase in industrial production field. The scheme is of important guidance for increasing the catalytic efficiency and specific activity of amylase and other enzymes.

The invention relates to a purifying method of recombinant human granulocyte colony stimulating factors and applies a fine separating step of opposite-phrase filling to a purifying technology of the recombinant human granulocyte colony stimulating factors for the first time. Compared with the prior art, the method adopts the opposite-phrase filling for the first time to finely separate the recombinant human granulocyte colony stimulating factors, the electrophoresis purity is increased to 99 percent from 90 percent, the HPLC purity is increased to 99 percent from 90 percent, and the specific activity is increased to 2.3*10U/mg from 6.1*10U/mg. The purifying technology of the recombinant human granulocyte colony stimulating factors is adopted to greatly increase the purity, the specific activity and the stability of the recombinant human granulocyte colony stimulating factors and can effectively reduce the clinical side effect of the recombinant human granulocyte colony stimulating factors.

The invention discloses a method for producing alpha interferon and a dedicated bacterial strain therefor. The bacterial strain is obtained by introducing the following genes into a host strain: (1) DNA molecules with the nucleotide sequence as shown in the sequence one in the sequence table; (2) DNA molecules whose functional protein is hybridized with and provided with the same coding with the DNA sequence as prescribed in (1) under strict conditions; and (3) DNA molecules whose functional protein has over 90% of homology with and has the same coding with the DNA sequence as prescribed in (1). The method for preparing the alpha interferon comprises the following step of fermenting the recombined strains to obtain the alpha interferon. The Escherichia coli DH5a/pBV220/Sw IFN-aCCMCC NO.2845 of the invention has good genetic stability, stable production performance, and the fermentation expressive recombined alpha interferon has high specific activity, high yield and high safety. The method of the invention is used for preparing the alpha interferon, thus the operation is simple, the cost is low and the efficiency is high. Therefore, the method for preparing alpha interferon of the invention has wide application prospect.

The invention provides a hemocoagulase extracted from bothrops atrox and also provides a preparation method of the hemocoagulase. The method comprises the following steps of: pretreating the bothropsatrox; sequentially passing the diethylaminoethyl cross-linked dextran gel A-25 ion exchange chromatography, benzamidine agarose gel 4-FF (HS) affinity chromatography, and cross-linked dextran gel G-75 gel filtration chromatography; and obtaining hemocoagulase. The invention also provides the application of the bothrops atrox hemocoagulase. in the preparation of a medicament for treating bleedingdisorder. Compared with the prior art, the preparation process steps are less, the period is short, and the exposure risk of the process environment is reduced; the product yield is high, the processhas strong ability to remove impurities, the yield of the product can be obtained more than 35% and the purity is over 98%; the process stability is high and the product quality is good, and the process is beneficial to clinical medication safety.