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38 results about "Serine proteinases" patented technology

Agkistrodon acutus hemocoagulase-B

The invention provides agkistrodon acutus hemocoagulase-B which is high-activity hemocoagulase separated from agkistrodon acutus venom. The hemocoagulase is single-stranded glycoprotein comprising 236 amino acids, and has an amino acid sequence shown as SEQ ID No. (sequence identification number) 1. The strand comprises 6 pairs of disulfide bonds; molecular weight of SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) is approximately 35kD; the molecular weight of the hemocoagulase after being deglycosylated is 26116.7Da; and an isoelectric point (pI) is 6.0. A hemocoagulase molecule is modified by heterozygotic polysaccharides at an Asn<77, 100, 229> (asparagine) site. The hemocoagulase is serine proteinase. The invention also provides a separation and purification method of the hemocoagulase, which comprises the steps that undissolved substances are removed through pretreating; then two times of anion-exchange column chromatography and one time of Sephdex-G75 molecular sieve chromatography are conducted; an active elution peak is acquired; dialysis and lyophilization are conducted; and the high-purity agkistrodon acutus venom hemocoagulase is obtained. The specific activity of the agkistrodon acutus venom hemocoagulase is not less than 2000U/mg; the analysis purity of HPLC (high performance liquid chromatography) can reach above 95%; and the yield is 0.25-0.30% calculated according to the weight of an agkistrodon acutus venom raw material.
Owner:BEIJING KONRUNS PHARM CO LTD

Expression optimization and purification method of helicobacter pylori serine proteinase

The invention discloses an expression optimization and purification method of helicobacter pylori serine proteinase. The expression optimization and purification method comprises the steps of constructing a target protein expression vector, performing induced expression of target protein under optimized condition, performing separation and purification on the target protein and the like. Accordingto the expression optimization and purification method disclosed by the invention, the target protein expression vector containing a His label is constructed and is successfully conveyed into host bacteria; besides, four factors of pre-inducing time, inducing temperature, the final concentration of an inducer and inducing time influencing growth of thalli of engineering bacteria and inducing expression quantity of the target protein are optimized through designing an orthogonal experiment, the optimal inducing expression condition is obtained, and the expression quantity of the target proteinis greatly increased; and an affinity chromatography method and an eluting system of the target protein are also optimized, so that the purity of the target protein reaches 98% and is notably increased than that by a traditional method, and the expression optimization and purification method has good application prospect on large-scale mass production of recombination protein.
Owner:GUANGDONG UNIV OF TECH
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