TITLE:
Molecular Cloning and Phylogenetic Analysis of a Chitin Deacetylase Isolated from the Epidermis of the Red Snow Crab Chionoecetes japonicas
AUTHORS:
Kakeru Fujimori, Hideto Fukushima, Masahiro Matsumiya
KEYWORDS:
Chitin Deacetylase, Molecular Cloning, Chionoecetes japonicas, Phylogenetic Analysis, Expression Analysis
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.9 No.1,
January
30,
2018
ABSTRACT: Chitin
deacetylase (CDA; EC 3. 5. 1. 41) catalyzes the deacetylation of chitin. In
this study, we successfully cloned and sequenced a chitin deacetylase gene from the red snow crab Chionoecetes japonicas. By
using reverse transcription-polymerase chain reaction (RT-PCR) and 5' and 3' rapid amplification of cDNA ends, we obtained a
2141-bp amplicon containing a chitin deacetylase gene (CjCDA) from
the epidermis of C. japonicas. The
amplicon contains
a 1575-bp open reading frame that is predicted to encode a 525-amino acid protein.
The structure predicted from the deduced amino acid sequence included an N-terminal signal peptide, chitin-binding
domain (CBD), low-density lipoprotein receptor class A domain (LDL-A),
and catalytic domain. Comparative analysis of the deduced amino acid sequence
of CjCDA revealed the highest homology (74%) to gastrolith protein 59 of Cherax quadricarinatus. We
used RT-PCR to evaluate the expression of CjCDA in
various tissues of C. japonicas, and
we observed that CjCDA was
expressed only in the epidermis. A phylogenetic analysis, using the amino acid
sequences of CjCDA and other known chitin deacetylases, showed that CjCDA
belonged to a group of crustacean chitin deacetylases. To our knowledge, this
is the first study reporting the cDNA cloning of a chitin deacetylase from a
crab.