########     ###    ##    ## ########  ######  
##     ##   ## ##   ###   ##    ##    ##    ## 
##     ##  ##   ##  ####  ##    ##    ##       
########  ##     ## ## ## ##    ##     ######  
##        ######### ##  ####    ##          ## 
##        ##     ## ##   ###    ##    ##    ## 
##        ##     ## ##    ##    ##     ######  

PETase ANnotation and Triage System

Nature's solution to a human-made health problem

Cut190**SS

304 aa · engineered from Cut190 · Oda et al. 2018, Appl. Microbiol. Biotechnol.

Disulfide-stabilised; calcium-dependent conformational switching

Structure, superposed on IsPETase

Aligned onto IsPETase 6EQE when it was written, so anything added below overlays directly and the browser does no alignment. The catalytic triad is drawn from the residues the geometry stage measured, not from positions inferred by an alignment.

Overlay another

Sequence 304 aa · 7 substitutions

catalytic triad   substitution against Cut190  · the same colours as the viewer above. Triad positions are labelled; hover any residue for its number. Click one to select it in both.

MRIRRQAGTGARASMARAIGVMTTALAVLVGAVGGVAGAEVSTAQDNPYERGPDPTEDSIEAIRGPFSVATERVSSFASGFGGGTIYYPRETDEGTFGAVAVAPGFTASQGSMSWYGERVASHGFIVFTIDTNTRLDAPGQRGRQLLAALDYLVERSDRKVRERLDPNRLAVMGH176SMGGGGSLEATVMRPSLKASIPLTPWHLDKTWGQVQVPTFIIGAEL222DTIAPVSTHAKPFYESLPSSLPKAYMELCGAT254HFAPNIPNTTIAKYVISWLKRFVDEDTRYSQFLCPNPTDRAICEYRSTCPY

Activity

Optimum temperature65.0 °C
As publishedPublished as 65 °C.
Optimum pHnot recorded
Familycutinase

Structure

Source Experimental, PDB 7CTR
Resolution1.2 Å
Residues259
Cα RMSD to IsPETase1.90 Å

Active site

Catalytic triadSer176 · His254 · Asp222
Ser OG → His NE22.73 Å
His ND1 → Asp OD3.22 Å
Oxyanion donor 1177 (3.03 Å)
Cleft width16.36 Å
Cleft depth4.15 Å
Cleft residues83

Aromatic clamp: PHE106 · PHE234 · PHE255 · TRP201

Mutations

Q123H, Q138A, N202H, S226P, R228S, D250C, E296C

7 substitutions against Cut190. Every one was applied by a routine that refuses any substitution whose stated parent residue does not match, so a wrong position or a mature-versus-precursor numbering shift fails loudly rather than producing a plausible but wrong sequence.

Disulfide-stabilised Cut190. CORRECTED after a literature check: this entry previously took its sequence from PDB 7CEF, which is Cut190* (S226P/R228S with a C-terminal truncation) and carries only the two native cysteines. The 'SS' in the name IS an engineered disulfide, so an entry without one was the wrong molecule under the right name. 7CTR is the real variant: 4 cysteines, and D250C/E296C form the added bridge. All seven published substitutions match Cut190 at offset 0 and take the cysteine count from 2 to 4, which is the check that settles it.

Reference: Oda et al. 2018, Appl. Microbiol. Biotechnol. doi:10.1007/s00253-018-9374-x

Measured activity

ParameterValueSubstrateEvidenceSource
performance claim PET from review 10.1007/s00253-018-9374-x
Disulfide-stabilised; calcium-dependent conformational switching
topt 65.0 degC PET from review 10.1007/s00253-018-9374-x
Published as 65 °C.

Related in PANTS

Lineage

Engineered from Cut190.

Nearest metagenomic candidates

No candidate in the catalogue names this enzyme as its nearest match.

Identifiers and cross-references

Disulfide-stabilised Cut190. CORRECTED after a literature check: this entry previously took its sequence from PDB 7CEF, which is Cut190* (S226P/R228S with a C-terminal truncation) and carries only the two native cysteines. The 'SS' in the name IS an engineered disulfide, so an entry without one was the wrong molecule under the right name. 7CTR is the real variant: 4 cysteines, and D250C/E296C form the added bridge. All seven published substitutions match Cut190 at offset 0 and take the cysteine count from 2 to 4, which is the check that settles it.