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Literature summary for 3.2.1.132 extracted from

  • Lyu, Q.; Wang, S.; Xu, W.; Han, B.; Liu, W.; Jones, D.N.; Liu, W.
    Structural insights into the substrate-binding mechanism for a novel chitosanase (2014), Biochem. J., 461, 335-345.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Microbacterium sp.

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with hexaglucosamine, hanging drop vapor diffusion method, using 0.05 M KH2PO4 and 23% (w/v) PEG 8000 using 0.7 M NaCl as the reservoir at 15°C Microbacterium sp.

Protein Variants

Protein Variants Comment Organism
D235A the mutant shows about 18% of wild type activity Microbacterium sp.
D43A inactive Microbacterium sp.
E25A inactive Microbacterium sp.
H203A the mutant shows about 3% of wild type activity Microbacterium sp.
S27A the mutant shows about 80% of wild type activity Microbacterium sp.
T58A the mutant shows about 20% of wild type activity Microbacterium sp.
Y37F the mutant shows about 5% of wild type activity Microbacterium sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
chitosan + H2O Microbacterium sp.
-
chitosan oligomers chitodimer and chitotrimer as main products, a monoacetyl chitodimer and a monoacetyl chitotrimer but no significant amount of monomer glucosamine or chitotetramer is identified ?
chitosan + H2O Microbacterium sp. OU01
-
chitosan oligomers chitodimer and chitotrimer as main products, a monoacetyl chitodimer and a monoacetyl chitotrimer but no significant amount of monomer glucosamine or chitotetramer is identified ?

Organism

Organism UniProt Comment Textmining
Microbacterium sp. A7KBW5
-
-
Microbacterium sp. OU01 A7KBW5
-
-

Purification (Commentary)

Purification (Comment) Organism
glutathione-Sepharose 4B bead chromatography, and Sephacryl S-100 gel filtration Microbacterium sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
chitohexaose + H2O
-
Microbacterium sp. ?
-
?
chitohexaose + H2O
-
Microbacterium sp. OU01 ?
-
?
chitosan + H2O
-
Microbacterium sp. chitosan oligomers chitodimer and chitotrimer as main products, a monoacetyl chitodimer and a monoacetyl chitotrimer but no significant amount of monomer glucosamine or chitotetramer is identified ?
chitosan + H2O
-
Microbacterium sp. OU01 chitosan oligomers chitodimer and chitotrimer as main products, a monoacetyl chitodimer and a monoacetyl chitotrimer but no significant amount of monomer glucosamine or chitotetramer is identified ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
42.5
-
melting temperature of the wild type enzyme Microbacterium sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
205
-
chitosan at pH 5.6 and 37°C Microbacterium sp.