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Literature summary for 3.2.1.14 extracted from

  • Kuttiyawong, K.; Nakapong, S.; Pichyangkura, R.
    The dual exo/endo-type mode and the effect of ionic strength on the mode of catalysis of chitinase 60 (CHI60) from Serratia sp. TU09 and its mutants (2008), Carbohydr. Res., 343, 2754-2762.
    View publication on PubMed
Search for more literature for 3.2.1.14

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha Serratia sp.

Protein Variants

Protein Variants Comment Organism
W245F site-directed mutagensis of the catalytic domain residue, the mutant shows reduced binding affinity towards substrate colloidal chitin Serratia sp.
W245Y site-directed mutagensis of the catalytic domain residue, the mutant shows reduced binding affinity towards substrate colloidal chitin Serratia sp.
W33F site-directed mutagensis of the N-terminal domain residue, the mutant shows reduced binding affinity towards substrate colloidal chitin Serratia sp.
W33F/W245F site-directed mutagensis of N-terminal domain and catalytic domain residues, the mutant shows a slight decrease in activity on beta-crystalline chitin, while demonstrating a slight increase in activity on colloidal chitin compared to the wild-type enzyme Serratia sp.
W33F/W69Y site-directed mutagensis of N-terminal domain residues, the mutant shows a slight decrease in activity on beta-crystalline chitin, while demonstrating a slight increase in activity on colloidal chitin compared to the wild-type enzyme Serratia sp.
W33Y site-directed mutagensis of the N-terminal domain residue, the mutant shows reduced binding affinity towards substrate colloidal chitin Serratia sp.
W69F site-directed mutagensis of the N-terminal domain residue, the mutant shows reduced binding affinity towards substrate colloidal chitin Serratia sp.
W69Y site-directed mutagensis of the N-terminal domain residue, the mutant shows reduced binding affinity towards substrate colloidal chitin Serratia sp.
W69Y/W245Y site-directed mutagensis of N-terminal domain and catalytic domain residues, the activity rate of mutant W69Y/W245Y is significantly higher than that of wild-type CHI60 on crystalline beta-chitin Serratia sp.

Organism

Organism UniProt Comment Textmining
Serratia sp. Q93LF1
-
-
Serratia sp. TU09 Q93LF1
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant enzymes by ultrafiltration, ion exchange chromatography and chitin affinity chromatography Serratia sp.

Reaction

Reaction Comment Organism Reaction ID
chitopentaose + H2O = chitobiose + chitotriose catalytic mechanism with an important role of Trp-33 on enzyme processivity, CHI60 preferentially uses the endo-type mode on soluble and amorphous substrates and the exo-type mode on crystalline substrate, the enzyme hydrolyzes crystalline substrates with an exo-type mode processively while remaining tightly bound to the substrate. The prevalent mode of hydrolysis mediated by CHI60 is regulated by ionic strength Serratia sp.
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 relative enzymatic activity of wild-type CHI60 and mutant enzymes at low and high ionic strengths on amorphous and crystalline substrates, overview Serratia sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
beta-chitin + H2O crystalline chitin Serratia sp. ?
-
 ?
beta-chitin + H2O crystalline chitin Serratia sp. TU09 ?
-
 ?
colloidal chitin + H2O amorphous chitin Serratia sp. ?
-
 ?
colloidal chitin + H2O amorphous chitin Serratia sp. TU09 ?
-
 ?
additional information CHI60 possesses a dual mode of catalysis with both exo- and endo-type activities allowing the enzyme to work efficiently on various substrate types Serratia sp. ?
-
 ?
additional information CHI60 possesses a dual mode of catalysis with both exo- and endo-type activities allowing the enzyme to work efficiently on various substrate types Serratia sp. TU09 ?
-
 ?
partially N-acetylated chitin + H2O soluble chitin Serratia sp. ?
-
 ?
partially N-acetylated chitin + H2O soluble chitin Serratia sp. TU09 ?
-
 ?

Synonyms

Synonyms Comment Organism
CHI60
-
 Serratia sp.
chitinase 60
-
 Serratia sp.