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Int J Biol Macromol ; 104(Pt B): 1648-1655, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28373044

RESUMO

Chitin is the second most abundant biopolymer in nature and is an important resource. In this study, we identified a chitinase gene, named Eisenia fetida-Chitinase (EF-Chi) gene, of 1494 base pairs (bp) that encodes a protein of 498 amino acids as indicated by the corresponding mRNA sequence. The amino acid sequence of EF-Chi was similar to those of chitinases from Eisenia andrei (99%), Branchiostoma floridae (50%) and Oryzias latipes (49%), and a gene encoding mature EF-Chi was expressed in the GS115 strain of Pichia pastoris. The molecular mass of the purified recombinant EF-Chi (rEF-Chi) was estimated to be 60kDa and catalytically important residues of chitinases of the glycoside hydrolase (GH) family 18 were conserved in EF-Chi. The optimal catalytic temperature of rEF-Chi was identified as 60°C, and the hydrolytic product from colloidal chitin was N-acetyl-chitobiose, suggesting that EF-Chi is an exo-type enzyme.


Assuntos
Quitinases/genética , Quitinases/metabolismo , Oligoquetos/enzimologia , Sequência de Aminoácidos , Animais , Quitinases/química , Clonagem Molecular , Estabilidade Enzimática , Expressão Gênica , Concentração de Íons de Hidrogênio , Hidrólise , Oligoquetos/genética , Filogenia , Pichia/genética , Especificidade por Substrato , Temperatura
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