Shandong University

Shandong Institute for Food and Drug Control

Avanc Pharmaceutical Co., Ltd

Bloomage Biotechnology Corporation Limited

Fengshan Wang

Feng Shi

Lianli Chi

Baojian Hang

Weijian Wang

Liping Gong

Ruiqing Xian

Congcong Wang

Xueping Guo

Hongming Du

Yuanjun Sun

Xunjie Zhang

Liu Sun

Li Nie

Ranran Du

Qunye Zhang

Juzheng Sheng

Jianqun Deng

Zhen Lu

A heparin skeleton synthase originates from, with an amino acid sequence as shown in SEQ ID NO.2 and a nucleotide sequence of the coding gene as shown in SEQ ID NO.1. Its recombinant expression level is 6.8 times that of the existing heparin skeleton synthase KfiA fromK5, and total enzyme activity per fermentation liquor is 5.22 times that of the heparin skeleton synthase KfiA. The heparin skeleton synthase mutants obtained through site-directed mutagenesis of the sites No. 16, No. 25, No. 30, No. 111, No. 165, and No. 172 in the amino acid sequence of the said heparin skeleton synthase all have high expression levels.

Heparin skeleton synthase and its mutants and application

The present invention provides a snake venom thrombin marker peptide ofand an application of the snake venom thrombin-like enzyme in identifying species of Hemocoagulase for Injection. The application includes the following steps of: dissolving a to-be-detected sample and a reference substance of the marker peptide respectively to prepare a test solution and a reference solution, and conducting alkylation reduction on the test solution and the reference solution with dithiothreitol and iodoacetamide; after diluting products with an ammonium bicarbonate solution, adding enzyme for hydrolysis; and after enzymolysis is finished, conducting centrifugation at a high speed, and injecting a supernatan into a liquid chromatography-mass spectrometer for analysis. This method is simple, convenient and rapid, is strong in specificity, fills the gap in identifying the source of species of the snake venom thrombin-like enzyme ofand improves the quality control level.

Snake venom thrombin-like enzyme marker peptide ofand its application in the species identification of hemocoagulase for injection

The present invention relates to the field of chemical analysis detection and application, in particular to a marker peptide of snake venom thrombin-like enzymes (SVTLEs) fromand an application thereof. The amino acid sequence of the marker peptide of snake venom thrombin-like enzymes (SVTLEs) fromis TLCAGVMEGGIDTCNR. Characterizing the source of species and a content of the SVTLEs in a to-be-detected sample by using the marker peptide includes the following steps of: pretreating the to-be-detected sample by trypsin through enzymolysis, and taking a supernatant of an enzymolysis liquid as a test solution; and injecting the test solution and a reference solution into a liquid chromatography-mass spectrometer, and selecting a qualitative ion pair and a quantitative ion pair for detecting the source of species and a content of the SVTLEs in the to-be-detected sample.

Marker peptide of snake venom thrombin-like enzymes (SVTLEs) from agkistrodon halys pallas and application thereof

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Shandong, China

Fengshan Wang