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Table 1 Proteases commonly used for protein hydrolysis

From: Protein hydrolysates in animal nutrition: Industrial production, bioactive peptides, and functional significance

Class of enzyme

Name of enzyme

EC number

Specific cleavage

Endopeptidases

 Aspartate protease

Chymosin (rennin; pH 1.8–2)

3.4.23.4

the Phe-Met bond, clotting of milk

Pepsin A (pH 1.8–2)

3.4.23.1

Aromatic AAs, hydrophobic AAs

 Cysteine protease

Bromelain (from pieapples)

3.4.22.4

Ala, Gly, Lys, Phe, Tyr

Cathepsin B

3.4.22.1

Arg, Lys, Phe

Ficain (ficin; from fig tree)

3.4.22.3

Ala, Asn, Gly, Leu, Lys, Tyr, Val

Papain (from papaya)

3.4.22.2

Arg, Lys, Phe

 Metallo protease

Bacillolysin (Bacillus bacteria)

3.4.24.28

Aromatic AAs, Ile, Leu, Val

Thermolysin (Bacillus bacteria)

3.4.24.27

Aromatic AAs, Ile, Leu, Val

 Serine protease

Chymotrypsin (pH 8–9)

3.4.21.1

Aromatic AAs, Leu

Subtilisin (from Bacillus bacteria)

3.4.21.14

Mainly hydrophobic AAs

Trypsin (pH 8–9)

3.4.21.4

Arg, Lys

Exopeptidases

 Aminopeptidases

Aminopeptidasea

3.4.11.1

AA at the N-terminus of protein/peptide

Aminopeptidase Yb

3.4.11.15

Lys at the N-terminus of protein/peptide

 Carboxypeptidase

Carboxypeptidasec

3.4.16.1

Acidic, neutral, and basic AAs

Glycine carboxypeptidased

3.4.17.4

Gly at the C-terminus of protein/peptide

Alanine carboxypeptidasee

3.4.17.8

D-Ala at the C-terminus of peptide

Carboxypeptidase Sf

3.4.17.9

Gly at the C-terminus of protein/peptide

 Dipeptidase

Dipeptidase 1f

3.4.13.11

A wide range of dipeptides

Proline dipeptidase (prolidase)a

3.4.13.9

AA-Pro or -hydroxyproline at the C-terminus (not Pro-Pro)

Prolyl dipeptidasea

3.3.13.8

Pro-AA or Hydroxyproline-AA

Endo- and exo-peptidases

 Pronase

A mixture of proteasesa (from Streptomyces griseus)

3.4.24

Acidic, neutral, and basic AAs

 Other peptidases

Dipeptidyl-peptide IIIf

3.4.14.4

Release of an N-terminal dipeptide from a peptide comprising four or more AA residues, with broad specificity

Dipeptidyl-peptidase IVg

3.4.14.5

Release of an N-terminal dipeptide from a peptide consisting of prolineh

  1. Adapted from Kunst [16] and Dixon and Webb [17]. AA amino acid
  2. aMetallopeptidase (requiring Mn2+, Mg2+ or Zn2+ for activation)
  3. bMetallopeptidase (requiring Co2+ for activation; inhibited by Zn2+ and Mn2+)
  4. cSerine carboxypeptidase
  5. dStrongly inhibited by Ag+ and Cu2+
  6. eMetallopeptidase (requiring Mn2+, Mg2+, Zn2+, Ca2+ or Co2+ for activation)
  7. fMetallopeptidase (requiring Zn2+ for activation)
  8. gSerie protease
  9. hAA1-Pro-AA2, where AA2 is neither proline nor hydroxyproline