The gene encoding fibrinolytic enzyme from Bacillus sp. zlw-2 was cloned and sequenced (accession no. EU734749), which was 1 146 bp, encoded 381 amino acids and had 99% homology with Nattokinase YF308 and NAT. The genes encoding pre-pro-fibrinolytic enzyme (including signal peptide, propeptide, and mature peptide) and fibrinolytic enzyme (including mature peptide) were cloned into pET28a vector respectively and then transformed into Escherichia coli BL21 (DE3). The recombinant of pre-pro-fibrinolytic enzyme showed enzyme activity of 183 U mL-1, while no detectable enzyme activity could be found from the recombinant of the mature peptide. The gene encoding fibrinolytic enzyme from Bacillus sp. Zlw-2 was cloned and sequenced (accession no. EU734749), which was 1 146 bp, encoded 381 amino acids and had 99% homology with Nattokinase YF308 and NAT. The genes encoding pre-pro -fibrinolytic enzyme (including signal peptide, propeptide, and mature peptide) and fibrinolytic enzyme (including mature peptide) were cloned into pET28a vector respectively and transformed into Escherichia coli BL21 (DE3). The recombinant of pre-pro-fibrinolytic enzyme showed enzyme activity of 183 U mL-1, while no detectable enzyme activity could be found from the recombinant of the mature peptide.