The thrombolytic effect of NKCP is attributable to a proteolytic enzyme named bacillopeptidase F, produced by Bacillus subtilis natto. Bacillopeptidase F is shown to be one of five proteases produced and secreted by Bacillus subtilis natto (Table 1).

Table 1: Enzymes extracellularly secreted by Bacillus subtilis

Protease	Gene	Properties
Bacillopeptidase F	bpr	Molecular weight by SDS-PAGE: 47kDa 1), 48kDa 2) Secreted as a 92kDa protein and converted into 80kDa and 48kDa proteins 2). It has high esterase activity as well as proteinase activity 3).
Subtilisn (alkaline) protease	apr	Molecular weight by SDS-PAGE: 20kDa4), 28kDa5) The structure is similar to that of nattokinase. Its casein decomposing activity and direct fibrinolytic activity have been confirmed. It is applied as protease in laundry detergent. The ability to decompose and inactivate plasminogen activator inhibitor type 1 (PAI-1) has also been reported 5).
Neutral protease	npr	A major protease like Subtilisin (alkaline) protease.
Extracellular protease	epr	Molecular weight by SDS-PAGE: 40-34kDa6)
Metallo protease	mpr	Molecular weight by SDS-PAGE: 28kDa1) Expression of the activity requires a metal atom.

1) Journal of Bacteriology 1990; 172: 1019-1023
2) The Journal of Biological Chemistry 1990; 265: 6845-6850
3) Journal of Bacteriology 1990; 172: 1470-1477
4) Experientia 1987; 43: 1110-1111
5) The Journal of Biological Chemistry 2001; 276: 24690-24696
6) Mol Gen Genet 1990 May; 221(3): 486-490

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