Back to All Enzymes

Subtiligase

Host Organism

Bacillus amyloliquefaciens

Type

Sequence

AQSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQDNNSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGADGSGQYSWIINGIEWAIANNMDVINMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAVDSSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPNWTNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ

External Links

PDB ID: -
EC: 3.4.21.62
MEROPS: S8 subtilase
UniProt ID: -
NCBI: -

Substrate Specificity

P2
P1
P1'
P1''
P2''
A/S/G
L/F/A/V
Ester/thioester
G/A/S
A/G

Functions

  • Intermolecular backbone terminal ligation
  • Backbone terminal cyclization
  • Specificity stringency: Prime-low, Nonprime-low, only stringent to the C-terminal active ester/thioester served as a leaving group

Expression Platform:

  • E. coli

Mutations:

  • Protease-to-ligase conversion of subtilisin BPN': S221C/P225A

Other Properties:

-

References:

  1. [1] (Discovery) Abrahmsén, L.; Tom, J.; Burnier, J.; Butcher, K. A.; Kossiakoff, A.; Wells, J. A. Engineering subtilisin and its substrates for efficient ligation of peptide bonds in aqueous solution. Biochemistry 1991, 30 (17), 4151-4159. DOI: 10.1021/bi00231a007
  2. [2] (Sequence, Activity) Chang, T. K.; Jackson, D. Y.; Burnier, J. P.; Wells, J. A. Subtiligase: a tool for semisynthesis of proteins. Proceedings of the National Academy of Sciences 1994, 91 (26), 12544-12548. DOI: 10.1073/pnas.91.26.12544
  3. [3] (Application) Braisted, A. C.; Judice, J. K.; Wells, J. A. Synthesis of proteins by subtiligase. Methods Enzymol 1997, 289, 298-313. DOI: 10.1016/s0076-6879(97)89053-2