Cloning and expression of a chitinase gene from Eisenia fetida

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 am...

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Bibliographic Details
Published in:International journal of biological macromolecules 2017-11, Vol.104 (Pt B), p.1648-1655
Main Authors: Ueda, Mitsuhiro, Shioyama, Takashi, Nakadoi, Kei, Nakazawa, Masami, Sakamoto, Tatsuji, Iwamoto, Takeo, Sakaguchi, Minoru
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Chitinase
Chitinases - chemistry
Chitinases - genetics
Chitinases - metabolism
Cloning, Molecular
Earthworm
Eisenia fetida
Enzyme Stability
Gene Expression
Glycoside hydrolase (GH) family 18
Hydrogen-Ion Concentration
Hydrolysis
Oligochaeta - enzymology
Oligochaeta - genetics
Phylogeny
Pichia - genetics
Substrate Specificity
Temperature
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Summary: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.
ISSN:0141-8130
1879-0003
DOI:10.1016/j.ijbiomac.2017.03.140