[Expression, purification and identification of human matrix metalloproteinase-2]

The expression sequence of matrix metalloproteinase-2 (MMP-2) has been obtained by PCR amplifying, restriction enzyme cut and sequencing analysis demonstrate that the sequence is correct. The recombinant expression plasmid pPIC9/MMP-2 containing MMP-2 is constructed and transformed the yeast Pichia pastoris. Recombinant matrix metalloproteinase-2 protein was expressed in Pichia pastoris in great deal after induction by methanol. The purity of the recombinant MMP-2 filtrated through Sephacryl S-200 reached to electrophoresis purity. With the ability to degrade gelatin and IV type collagen, recombinant MMP-2 has the similar substrate specificity with natural MMP-2. The recombinant MMP-2 with 50 kD molecular weight is smaller than natural MMP-2, which suggested degradation occurred to it.