ABB  Vol.8 No.10 , October 2017
Distribution of Chitinolytic Enzymes in the Organs and cDNA Cloning of Chitinase Isozymes from the Liver of Golden Cuttlefish Sepia esculenta
ABSTRACT
The distribution of chitinolytic enzymes in eight organs of the golden cuttlefish Sepia esculenta was determined. Chitinase activity (activity of endo-type chitinolytic enzyme) was measured using pNP-(GlcNAc)n (n = 2, 3) as substrates, with high activity detected in the liver, posterior salivary gland, and stomach. β-N-acetylhexosaminidase (Hex) activity (activity of exo-type chitinolytic enzyme) was determined using pNP-(GlcNAc) as a substrate, and high activity was observed in six organs, including the liver, branchial heart, posterior salivary gland, and stomach. In addition, two chitin-binding proteins (CBP-A, CBP-B) were isolated from the liver using a chitin affinity column. Two full-length cDNAs (SeChi-1: 1484 bp; SeChi-2: 1748 bp) encoding chitinases were obtained from the liver of S. esculenta. SeChi-1 contained a 1377-bp open reading frame (ORF) encoding 459 amino acids, and SeChi-2 contained a 1656-bp ORF encoding 552 amino acids. Domain structures predicted from the deduced amino acid sequences of SeChi-1 and SeChi-2 (SeChi-1, SeChi-2) contained signal peptides, a GH Family 18 catalytic domain, one chitin binding domain (CBD) in SeChi-1, and two CBDs in SeChi-2. Proteome analysis revealed that 125 peptide residues of CBP-A were present in SeChi-1, and 116 peptide residues of CBP-B were present in SeChi-2. Organ expression analysis revealed that SeChi-1 and SeChi-2 were expressed only in the liver of S. esculenta. Phylogenetic analysis of SeChi-1, SeChi-2, and GH family 18 chitinases revealed that SeChi-2 belongs to a group of previously reported squid chitinases, while SeChi-1 does not belong to any previously reported group of mollusk chitinases.
Cite this paper
Nishino, R. , Kakizaki, H. , Fukushima, H. and Matsumiya, M. (2017) Distribution of Chitinolytic Enzymes in the Organs and cDNA Cloning of Chitinase Isozymes from the Liver of Golden Cuttlefish Sepia esculenta. Advances in Bioscience and Biotechnology, 8, 361-377. doi: 10.4236/abb.2017.810026.
References
[1]   Arbia, W., Arbia, L., Adour, L. and Amrane, A. (2013) Chitin Extraction from Crustacean Shells Using Biological Methods—A Review. Food Technology and Biotechnology, 51, 12-25.

[2]   Karthik, N., Akanksha, K., Binod, P. and Pandey, A. (2014) Production, Purification and Properties of Fungal Chitinases—A Review. Indian Journal of Experimental Biology, 52, 1025-1035.

[3]   Komatsu, M., Son, J., Matsushita, N. and Hogetsu, T. (2007) Fluorescein-Labeled Wheat Germ Agglutinin Stains the Pine Wood Nematode, Bursaphelenchusxyiophilus. Journal of Forest Research, 13, 132-136.
https://doi.org/10.1007/s10310-008-0065-9

[4]   Khoushab, F. and Yamabhai, M. (2010) Chitin Research Revisited. Marine Drugs, 8, 1988-2012.
https://doi.org/10.3390/md8071988

[5]   Patil, R.S., Ghormade, V.V. and Deshpande, M.V. (2000) Chitinolytic Enzymes: An Exploration. Enzyme and Microbia Technology, 26, 473-483.
https://doi.org/10.1016/S0141-0229(00)00134-4

[6]   Slámová, K., Bojarová, P., Petrásková, L. and Kren, V. (2010) β-N-acetylhexosaminidase: What’s in a Name…? Biotechnology Advances, 28, 682-693.
https://doi.org/10.1016/j.biotechadv.2010.04.004

[7]   Gooday, G.W. (1999) Aggressive and Defensive Roles for Chitinases. Cellular and Molecular Life Sciences, 87, 157-169.

[8]   Henrissat, B. and Bairoch, A. (1993) New Families in the Classification of Glycosyl Hydrolases Based on Amino Acid Sequence Similarities. Biochemical Journal, 293, 781-788.
https://doi.org/10.1042/bj2930781

[9]   Fukamizo, T., Koga, D. and Goto, S. (1995) Comparative Biochemistry of Chitinases-Anomeric Form of the Reaction Products. Bioscience Biotechnology Biochemistry, 59, 311-313.
https://doi.org/10.1271/bbb.59.311

[10]   Koga, D., Yoshioka, K. and Arakane, Y. (1998) HPLC analysis of Anomeric Formation and Cleavage Pattern by Chitinolytic Enzyme. Bioscience Biotechnology Biochemistry, 62, 1643-1646.
https://doi.org/10.1271/bbb.62.1643

[11]   Shinya, S., Nagata, T., Ohnuma, T., Taira, T., Nishimura, S. and Fukamizo, T. (2012) Backbone Chemical Shifts Assignments, Secondary Structure, and Ligand Binding of a Family GH-19 Chitinase from Moss, Bryumcoronatum. Biomolecular NMR Assignments, 2, 157-161.
https://doi.org/10.1007/s12104-011-9346-x

[12]   Ikeda, M., Miyauchi, K., Mochizuki, A. and Matsumiya, M. (2009) Purification and Characterization of Chitinase from the Stomach of Silver Croaker Pennahiaargentatus. Protein Expression and Purification, 65, 214-222.

[13]   Ikeda, M., Miyauchi, K. and Matsumiya, M. (2012) Purification and Characterization of a 56 kDa Chitinase Isozyme (PaChiB) from the Stomach of Silver Croakerpennahiaargentatus. Bioscience Biotechnology Biochemistry, 76, 971-979.
https://doi.org/10.1271/bbb.110989

[14]   Kurokawa, T., Uji, S. and Suzuki, T. (2004) Molecular Cloning of Multiple Chitinase Genes in Japanese Flounder, Paralichthysolivaceus. Comparative Biochemistry and Physiology B, 138, 255-264.

[15]   Ikeda, M., Kondo, Y. and Matsumiya, M. (2013) Purification, Characterization, and Molecular Cloning of Chitinases from the Stomach of the Threeline Grunt Parapristipomatrilineatum. Process Biochemistry, 48, 1324-1334.

[16]   Kakizaki, H., Ikeda, M., Fukushima, H. and Matsumiya, M. (2015) Distribution of Chitinolytic Enzymes in the Organs and cDNA Cloning of Chitinase Isozymes from the Stomach of Two Species of Fish, Chub Mackerel (Scomber japonicus) and Silver Croaker (Pennahiaargentata). Open Journal of Marine Science, 5, 398-411.
https://doi.org/10.4236/ojms.2015.54032

[17]   Kawashima, S., Ikehata, H., Tada, C., Ogino, T., Kakizaki, H., Ikeda, M., Fukushima, H. and Matsumiya, M. (2016) Stomach Chitinase from Japanese Sardine Sardinopsmelanostictus: Purification, Characterization, and Molecular Cloning of Chitinase Isozymes with a Long Linker. Marine Drugs, 14, 1-13.
https://doi.org/10.3390/md14010022

[18]   Matsumiya, M. Kakizaki, H. and Ikeda, M. (2017) Prosperity Strategy and Chitinase in Fish. Chitin and Chitosan Research, 23, 4-16.

[19]   Ikeda, M., Kakizaki, H. and Matsumiya, M. (2017) Biochemistry of Fish Stomach Chitinase. International Journal of Biological Macromolecules, 104, 1672-1681.

[20]   Wang, G.L., Xu, B., Bai, Z.Y. and Li, J.L. (2012) Two Chitin Metabolic Enzyme Genes from Hyriopsiscumingii: Cloning, Characterization, and Potential Functions. Genetics and Molecular Research, 11, 4539-4551.
https://doi.org/10.4238/2012.October.15.4

[21]   Yonezawa, M., Sakuda, S., Yoshimura, E. and Suzuki, M. (2016) Molecular Cloning and Functional Analysis of Chitinases in the Fresh Water Snail, Lymnaeastagnalis. Journal of Structural Biology, 196, 107-118.

[22]   Li, H., Wang, D., Denq, Z., Huang, G., Fan, S., Zhou, D., Liu, B., Zhang, B. and Yu, D. (2017) Molecular Characterization and Expression Analysis of Chitinase from the Pearl Oyster Pinctadafucata. Comparative Biochemistry and Physiology Part B: Biochemistry & Molecular Biology, 203, 141-148.

[23]   Badariotti, F., Lelong, C., Dubos, M.P. and Favrel, P. (2007) Characterization of Chitinase-Like Proteins (Cg-Clp1 and Cg-Clp2) Involved in Immune Defence of the Mollusk Crassostreagigas. The FEBS Journal, 274, 3646-3654.
https://doi.org/10.1111/j.1742-4658.2007.05898.x

[24]   Badariotti, F., Thuau, R., Lelong, C., Dubos, M.P. and Favrel, P. (2007) Characterization of an Atypical Family 18 Chitinase from the Oyster Crassostreagigas: Evidence for a Role in Early Development and Immunity. Developmental & Comparative Immunology, 31, 559-570.

[25]   Gao, L., Xu, G. J., Su, H., Gao, X.G., Li, Y.F., Bao, X.B., Liu, W.D. and He, C.B. (2014) Identification and Expression Analysis of cDNA Encoding Chitinase-Like Protein (CLP) Gene in Japanese Scallop Mizuhopectenyessoensis. Genetics and Molecular Research, 13, 10727-10740.
https://doi.org/10.4238/2014.December.18.14

[26]   Matsunaga, G., Karasuda, S., Nishino, R., Fukushima, H. and Matsumiya, M. (2016) Molecular Cloning of a Chitinase Gene from the Ovotestis of Kuroda’s Sea Hare Aplysiakurodai. Advances in Bioscience and Biotechnology, 7, 38-46.
https://doi.org/10.4236/abb.2016.71005

[27]   Yang, B., Zhang, M., Li, L., Pu, F., You, W. and Ke, C. (2015) Molecular Analysis of Atypical Family 18 Chitinase from Fujian Oyster Crassostreaangulata and Its Physiological Role in the Digestive System. PLoS ONE, 10, e0129261.
https://doi.org/10.1371/journal.pone.0129261

[28]   Matsumiya, M. and Mochizuki, A. (1997) Purification and Characterization of Chitinase from the Liver of Japanese Common Squid Todarodespacificus. Fisheries Science, 63, 409-413.
https://doi.org/10.2331/fishsci.63.409

[29]   Matsumiya, M., Miyauchi, K. and Mochizuki, A. (2002) Characterization of 38kDa and 42kDa Chitinase Isozymes from the Liver of Japanese Common Squid Todarodespacificus. Fisheries Science, 68, 603-609.
https://doi.org/10.1046/j.1444-2906.2002.00467.x

[30]   Matsumiya, M., Miyauchi, K. and Mochizuki, A. (2003) Purification and Some Properties of a Chitinase Isozyme from the Liver of Japanese Common Squid Todarodespacificus. Fisheries Science, 69, 427-429.
https://doi.org/10.1046/j.1444-2906.2003.00640.x

[31]   Nishino, R., Suyama, A., Ikeda, M., Kakizaki, H. and Matsumiya, M. (2014) Purification and Characterization of a Liver Chitinase from Golden Cuttlefish, Sepia esculenta. Journal of Chitin and Chitosan Science, 2, 238-243.
https://doi.org/10.1166/jcc.2014.1065

[32]   Kondo, H., Morita, T., Ikeda, M., Kurosaka, C., Shitara, A., Honda, Y., Nozaki, R., Aoki, T. and Hirono, I. (2010) Identification of Enzyme Genes in the Liver of the Bleeker’s Squid Loligobleekeri by Expressed Sequence Tag Analysis. Fisheries Science, 76, 161-165.
https://doi.org/10.1007/s12562-009-0182-1

[33]   Fry, B.G., Roelants, K. and Norman, J.A. (2009) Tentacles of Venom: Toxic Protein Convergence in the Kingdom Animalia. Journal of Molecular Evolution, 68, 311-321.
https://doi.org/10.1007/s00239-009-9223-8

[34]   Ruder, T., Sunagar, K., Undheim, E.A., Ali, S.A., Wai, T.C., Low, D.H., Jackson, T.N., King, G.F., Antunes, A. and Fry, B.G. (2013) Molecular Phylogeny and Evolution of the Proteins Encoded by Coleoid (Cuttlefish, Octopus, Squid) Posterior Venom Glands. Journal of Molecular Evolution, 76, 192-204.
https://doi.org/10.1007/s00239-013-9552-5

[35]   Kremer, N., Philipp, E.E., Carpentier, M.C., Brennan, C.A., Kraemer, L., Altura, M.A., Augustin, R., Häsler, R., Heath-Heckman, E.A., Peyer, S.M., Schwartzman, J., Rader, B.A., Ruby, E.G., Rosenstiel, P. and McFall-Ngai, M.J. (2013) Initial Symbiont Contact Orchestrates Host-Organ-Wide Transcriptional Changes that Prime Tissue Colonization. Cell Host & Microbe, 14, 183-194.

[36]   Alves, D.M., Cristo, M., Sendao, J. and Borges, T.C. (2006) Diet of the Cuttlefish Sepia officinalis (Cephalopoda: Seplidae) off the South Coast of Portugal (Eastern Algarve). Journal of the Marine Biological Association of the United Kingdom, 86, 429-436.
https://doi.org/10.1017/S0025315406013312

[37]   Matsumiya, M., Miyauchi, K. and Mochizuki, A. (1998) Distribution of Chitinase and β-N-Acetylhexosaminidase in the Organs of a Few Squid and a Cuttlefish. Fisheries Science, 64, 166-167.
https://doi.org/10.2331/fishsci.64.166

[38]   Ohtakara, A. (1988) Chitinase and β-N-acetylhexosaminidase from Pycnoporuscinnabarinus. Method in Enzymology, 161, 462-470.

[39]   Olsen, J.V., Ong, S.E. and Mann, M. (2004) Trypsin Cleaves Exclusively C-Terminal to Arginine and Lysine Residues. Molecular & Cellular Proteomics, 3, 608-614.
https://doi.org/10.1074/mcp.T400003-MCP200

 
 
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