Supporting information:

Figure S1. Elution profile of 19 different amino acids by PPSQ-33A automated protein

and peptide sequencer: Aspatic acid (Asp), glutamic acid (Glu), asparagine(Asn),

glutamine (Gln), serine (Ser), threonine (Thr),histidine (His), glycine(Gly), alanine

(Ala), tyrosine (Tyr), Arginine (Arg), methionine (met), valine (Val), proline (Pro),

tryptophan (Trp), phenylalanine (Phe), lysine (Lys), isoleucine (ile), and leucine (Leu).

Observed by product of sequencing such as N, N’-diphenylthiourea (DPTU).

Figure S2. The first 6 N-terminal amino acids sequence of the purified enzyme analyzed

by PPSQ-33A Automatic protein and peptide sequencer after calibrated with the PTH-

amino acids standard. According to the comparison with standard, the N-terminal amino

acids sequence of the purified enzyme is NH2-Ala-Gln-Ser-Val-Pro-Tyr-Gly-Ile-Ser-

Gln, which is consistent with the N-terminal sequence of nattokinase.

Figure S3. The 7th -10th N-terminal amino acids sequence of the purified enzyme

analyzed by PPSQ-33A Automatic protein and peptide sequencer after calibrated with

the PTH-amino acids standard. According to the comparison with standard, the N-

terminal amino acids sequence of the purified enzyme is NH2-Ala-Gln-Ser-Val-Pro-Tyr-

Gly-Ile-Ser-Gln, which is consistent with the N-terminal sequence of nattokinase.

Graphical abstract