The invention discloses novel heparinase CMHep obtained from Chryseobacterium meningosepticum (Chryseobacterium meningosepticum), which is not reported. The heparinase is prepared by the following steps: performing fermentation culture on the screened bacteria, performing crude enzyme extraction, performing multi-step column chromatography and the like. The property study proves that the heparinase has specificity in the aspect of enzymolysis of heparin and is expected to be used for the aspect of low molecular heparin preparation or heparin quality detection.