A production method of lumbrokinase is characterized by comprising the following steps: (1) putting fresh lumbrokinase into a basket or cleaning it on the ground, removing the soil layer on the upper part of the earthworm, spreading the earthworm into a thin layer, illuminating for 10- 18 hours, allowing the earthworm to discharge the dirt, washing it twice with tap water, and washing off the soil and sticky floating objects. (2) grinding earthworm into slurry with colloid mill, and homogenizing with sodium citrate with pH of 5.0-5.3 and low concentration ethanol according to the ratio of (v/v)1:3-5; (3) slag removal: coarse slag in the feed liquid is removed by a settling centrifuge to prepare supernatant; (4) adding ammonium sulfate into the salting-out supernatant according to 30% saturation concentration, stirring until it is completely dissolved, and standing for 65438+/-0 hours; (5) removing fine slag from the feed liquid by a vertical high-speed centrifuge, and performing high-speed centrifugation to obtain clear liquid; (6) ultrafiltering the supernatant with a 50,000,000 dalton membrane to obtain ultrafiltrate; (7) column chromatography separation: pumping the ultrafiltrate into a discharge barrel, sampling after mixing, washing the ultrafiltrate with a washing solution until the effluent is light yellow with a pH of 7.0-8.0 and slightly alkaline, and then eluting with an eluent, so that the eluent becomes turbid and the color changes from yellow to red; When the conductivity of the effluent is slightly higher than that of the eluent, it starts to collect and elute until the feed is clear and the color is light orange, and the conductivity reaches that of the eluent. (8) Ultrafiltration: pour the collected liquid into the storage tank, start the pump, adjust the rotating speed to 265,438+0× 65,438+02 rpm, adjust the pressure to not more than 0.65,438+0 MPa, concentrate to 65,438+0/65,438+05, and then clarify the collected liquid. (9) filtering and sterilizing: filtering for three times with a clear sterile liquid net filter plate until the filtrate is clear, and sterilizing and filtering with a 0.22um microporous filter membrane with a grade of 300,000; (10) spray drying was carried out in a drying tower, the outlet temperature was controlled at 80-86℃, the feed liquid was dried, and enzyme powder was collected.
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