3.4.11.B8	Ala-Ala + H2O	-	Pyrococcus horikoshii	Ala + Ala	-	?	36448	
3.4.11.B8	Ala-Ala + H2O	-	Pyrococcus horikoshii OT-3	Ala + Ala	-	?	36448	
3.4.11.B8	Ala-Ala 4-nitroanilide + H2O	the tetracosameric complex shows maximal activity against Ala-Ala-Ala and Ala-Ala 4-nitroanilide	Pyrococcus horikoshii	Ala + Ala 4-nitroanilide	-	?	443021	
3.4.11.B8	Ala-Ala-Ala + H2O	the tetracosameric complex shows maximal activity against Ala-Ala-Ala and Ala-Ala 4-nitroanilide	Pyrococcus horikoshii	Ala + Ala-Ala	-	?	364820	
3.4.11.B8	Ala-Ala-Ala + H2O	the tetracosameric complex shows maximal activity against Ala-Ala-Ala and Ala-Ala-4-nitroanilide	Pyrococcus horikoshii	Ala + Ala-Ala	-	?	364820	
3.4.11.B8	Ala-Ala-Ala + H2O	the tetracosameric complex shows maximal activity against Ala-Ala-Ala and Ala-Ala 4-nitroanilide	Pyrococcus horikoshii OT-3	Ala + Ala-Ala	-	?	364820	
3.4.11.B8	Ala-Ala-Ala + H2O	the tetracosameric complex shows maximal activity against Ala-Ala-Ala and Ala-Ala-4-nitroanilide	Pyrococcus horikoshii OT-3	Ala + Ala-Ala	-	?	364820	
3.4.11.B8	Ala-Ala-Ala 4-nitroanilide + H2O	best substraet for the dodecameric form of the enzyme	Pyrococcus horikoshii	Ala + Ala 4-nitroanilide + Ala-Ala 4-nitroanilide	-	?	443022	
3.4.11.B8	Ala-Ala-Ala 4-nitroanilide + H2O	best substrat for the dodecameric form of the enzyme	Pyrococcus horikoshii	Ala + Ala 4-nitroanilide + Ala-Ala 4-nitroanilide	-	?	443022	
3.4.11.B8	Asp-Ala + H2O	-	Pyrococcus horikoshii	Asp + Ala	-	?	94904	
3.4.11.B8	Asp-Ala + H2O	-	Pyrococcus horikoshii OT-3	Asp + Ala	-	?	94904	
3.4.11.B8	additional information	PhTET1 activity cannot be detected by using chromogenic aminoacyl compounds (7-amino-4-methylcoumarin (AMC) or p-nitroanilide), probably because the X-AMC and X-4-nitroanilide molecules cannot fit into the active site pocket or because these compounds fit but are not hydrolyzed. HPLC analysis of Ala-Ala-p-nitroanilide degradation by the two PhTET1 oligomers shows that the generation of Ala-4-nitroanilide is not linear with time, suggesting product inhibition. PhTET1 can cleave the N-terminal amino acid of larger peptides, even when the 4-nitroanilide moiety is in the C-terminal position, and can also hydrolyze dipeptides like Ala-Ala or Asp-Ala. Therefore, PhTET1 seems to be unable to cleave a peptidic bond when 4-nitroanilide or 7-amino-4-methylcoumarin is in the P1' position, making activity assays difficult. Activity of the dodecamer enzyme form increases with substrate chain length. No hydrolytic activity toward N-acetyl-Leu-4-nitroanilide and N-succinyl-Ala-Ala 4-nitroanilide	Pyrococcus horikoshii	?	-	?	89	
3.4.11.B8	additional information	PhTET1 activity cannot be detected by using chromogenic aminoacyl compounds (7-amino-4-methylcoumarin (AMC) or p-nitroanilide), probably because the X-AMC and X-4-nitroanilide molecules cannot fit into the active site pocket or because these compounds fit but are not hydrolyzed. HPLC analysis of Ala-Ala-p-nitroanilide degradation by the two PhTET1 oligomers shows that the generation of Ala-4-nitroanilide is not linear with time, suggesting product inhibition. PhTET1 can cleave the N-terminal amino acid of larger peptides, even when the 4-nitroanilide moiety is in the C-terminal position, and can also hydrolyze dipeptides like Ala-Ala or Asp-Ala. Therefore, PhTET1 seems to be unable to cleave a peptidic bond when 4-nitroanilide or 7-amino-4-methylcoumarin is in the P1' position, making activity assays difficult. Activity of the dodecamer enzyme form increases with substrate chain length. No hydrolytic activity toward N-acetyl-Leu-4-nitroanilide and N-succinyl-Ala-Ala 4-nitroanilide	Pyrococcus horikoshii OT-3	?	-	?	89