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Literature summary extracted from

  • Chen, L.; Liu, T.; Zhou, Y.; Chen, Q.; Shen, X.; Yang, Q.
    Structural characteristics of an insect group I chitinase, an enzyme indispensable to moulting (2014), Acta Crystallogr. Sect. D, 70, 932-942.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.14 overexpression of C-terminally His6-tagged wild-type and mutant enzymes in Pichia pastoris strain GS115 and secretion to the culture medium Ostrinia furnacalis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.14 purified recombinant His6-tagged wild-type enzyme and mutant E148Q unliganded, and wild-type enzyme and mutant E148A in complex with chitobiose and /or chitotriose, hanging drop vapour diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 50 mM HEPES, pH 8.0, and 100 mM NaCl, with 0,001 ml of reservoir solution containing 100 mM HEPES, pH 7.5, 25% w/v PEG 3350 for the wild-type with soaking of crystals in ligand containing solution for the complex crystals, reservori solution containing 5 mM (GlcNAc)6 in 100 mM HEPES, pH 7.9, 23% w/v PEG 3350 and in 100 mM HEPES, pH 8.0, 21% w/v PEG 3350, respectively, for mutant E148A and E148Q, X-ray diffraction structure determination and analysis at 1.7 A and 2.2 A resolution Ostrinia furnacalis

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.14 E148A site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 E148Q site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 F159A site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 F159A/F194A/W241A/Y290A site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 F159A/Y290A site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 F194A site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 F194A/W241A site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 W241A site-directed mutagenesis Ostrinia furnacalis
3.2.1.14 Y290A site-directed mutagenesis Ostrinia furnacalis

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.14 Ostrinia furnacalis Q2V6H4
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Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.14 recombinant His6-tagged wild-type and mutant enzymes from Pichia pastoris strain GS115 culture supernatant by ammonium sulfate fractionation and nickel affinity chromatography Ostrinia furnacalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.14 chitin + H2O the enzyme acts from non-reducing ends. Substrate binding structure, overview. The reducing sugar at subsite -1 is in an energetically unfavoured boat conformation, a state that possibly exists just before the completion of catalysis. A hydrophobic plane composed of four surface-exposed aromatic residues is adjacent to the entrance to the substrate-binding cleft Ostrinia furnacalis ?
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 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.14 group I chitinase
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 Ostrinia furnacalis
3.2.1.14 OfChtI
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 Ostrinia furnacalis

General Information

EC Number General Information Comment Organism
3.2.1.14 evolution the enzyme belongs to the glycosyl hydrolase family 18 (GH18) of chitinases Ostrinia furnacalis
3.2.1.14 additional information catalytic domain structure with and without bound substrate, structure comparisons and structure-function relationship, overview Ostrinia furnacalis
3.2.1.14 physiological function the enzyme is essential to moulting Ostrinia furnacalis