Chondrichthyes Chitinase: Molecular Cloning, Distribution, and Phylogenetic Analysis
- 1 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 2 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 3 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 4 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 5 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 6 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 7 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 8 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 9 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 10 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 11 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
- 12 Laboratory of Biocycle Engineering, Department of Applied Biological Chemistry, Osaka Prefecture University, Sakai, Japan
- 13 College of Bioresource Sciences, Nihon University, Fujisawa, Japan
Abstract
We have previously reported the presence of three types of chitinase (acidic fish chitinase-1: AFCase-1, acidic fish chitinase-2: AFCase-2, fish chitinase-3: FCase-3) in Actinopterygii. In the present research, we report the identification of the novel chitinase genes HjChi (ORF: 1380 bp) and DkChi (ORF: 1440 bp) from the stomach of Chondrichthyes, Japanese bullhead shark ( Heterodontus japonicas ) and Kwangtung skate ( Dipturus kwangtungensis ), respectively. Organ-specific expression analysis identified the stomach-specific expression of HjChi , whereas DkChi was expressed widely in all organs. Chitinase activity was measured using p NP-(GlcNAc) n ( n = 2, 3) as a substrate and β-N -acetylhexosaminidase (Hex) activity was measured using p NPGlcNAc. Relatively high values of chitinase activity were observed in the stomach, spleen, and gonads of the Japanese bullhead shark, H. japonicas , compared with that observed in the stomach of the Kwangtung skate D. kwangtungensis . However, Hex activity was detected throughout the body of both species. The optimal pH of chitinase in both the Japanese bullhead shark, H. japonicas , and the Kwangtung skate, D. kwangtungensis , were 3.5 - 5.5 and 3.5 - 4.0, respectively, and 4.0 for Hex in both species. Phylogenetic analysis revealed that Chondrichthyes chitinase forms a unique group (Chondrichthyes chitinase). These results suggested that the possibility of the formation of chitinase groups for each class in the phylogenetic analysis based on the observation of class-specific chitinase.
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