The invention relates to single chain protein matter with coagulation activity. The gel electrophoresis of the reducing SDS-polyacrylamide of the protein matter is performed to form a single strip under the molecular weight of 31+/-2kDa, the molecular weight is measured to be 30500+/-1000Da by a MALDI-TOF method, the isoelectric point is 4.4-4.8, the area normalization method is utilized to measure the purity which is more than 98%, and fifteen-amino acid sequence of N terminal end measured by the Edman degradation method is VIGGNECDTNEHRFL. The protein matter has stronger coagulation activity on human fibrinogen in vitro, and the specific activity thereof is 500-1500U/mg. Verified by in vitro coagulation activity experiment tests, the single chain protein matter with coagulation activityprovided by the invention has better coagulation activity; the preparation method thereof utilizes frequently-used separation and purification measures in the extraction separation process of currentbiochemical filed, thereby having strong process maneuverability, simple and easy operation and stable volume of production.