Skip to main content
SpringerLink
Log in

Partial characterization and ontogenetic development of pancreatic digestive enzymes in Japanese eel Anguilla japonica larvae

  • Published:
Fish Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

The pancreatic digestive enzymes, trypsin, chymotrypsin, lipase and amylase were partially characterized, and changes in their activities were examined during the initial ontogeny of Japanese eel Anguilla japonica larvae from 5 to 34 days post-hatching (dph). The pH optima of the eel larval enzymes were narrower than those other fish species; trypsin activity was highest at pH 9, chymotrypsin and amylase activities were highest at pH 7 and 8, and lipase activity was highest at pH 8 and 9. In an analysis of thermal profiles, the larval pancreatic enzymes had a high optimal temperature and high thermal stability, which are typical of fish from the tropics. At 12 and 13 dph, lipase activity and gene expression levels of trypsin (-a and -b), lipase and amylase decreased markedly, suggesting a marked change in larval metabolism at that time. These data could be useful in the development of artificial larval diets in Japanese eel.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Albro PW, Hall RD, Corbett JT, Schroeder J (1985) Activation of nonspecific lipase (EC 3.1.1.-) by bile salts. Biochim Biophys Acta 835:477–490

    Article  CAS  PubMed  Google Scholar 

  • Alvarez-González CA, Cervantes-Trujano M, Tovar-Ramírez D, Conklin D, Nolasco H, Gisbert E, Piedrahita R (2006) Development of digestive enzymes in California halibut Paralichthys californicus larvae. Fish Physiol Biochem 31:83–93

    Google Scholar 

  • Borlongan IG (1990) Studies on the digestive lipases of milkfish, Chanos chanos. Aquaculture 89:315–325

    Article  CAS  Google Scholar 

  • Cao M, Osatomi K, Suzuki M, Hara K, Tachibana K, Ishihara T (2000) Purification and characterization of two anionic trypsins from the hepatopancreas of carp. Fish Sci 66:1172–1179

    Article  CAS  Google Scholar 

  • Caruso G, Genovese L, Micale V, Spedicato MT, Mancuso M (2001) Preliminary investigation of the digestive enzymes in Pagellus erythrinus (Linneo 1758) larvae. Mar Fresh Behav Physiol 34:265–268

    Article  CAS  Google Scholar 

  • Castillo-Yáñez FJ, Pacheco-Aguilar R, Lugo-Sanchez ME, Garcia-Sanchez G, Quintero-Reyes IE (2009) Biochemical characterization of an isoform of chymotrypsin from the viscera of Monterey sardine (Sardinops sagax caerulea), and comparison with bovine chymotrypsin. Food Chem 112:634–639

    Article  Google Scholar 

  • Chen B, Qin J, Kumar M, Hutchinson W, Clarke S (2006) Ontogenetic development of digestive enzymes in yellowtail kingfish Seriola lalandi larvae. Aquaculture 260:264–271

    Article  CAS  Google Scholar 

  • Chiu S, Pan B (2002) Digestive protease activities of juvenile and adult eel (Anguilla japonica) fed with floating feed. Aquaculture 205:141–156

    Article  CAS  Google Scholar 

  • Darias MJ, Murray HM, Gallant JW, Astola A, Douglas SE, Yúfera M, Martínez-Rodríguez G (2006) Characterization of a partial alpha-amylase clone from red porgy (Pagrus pagrus): expression during larval development. Comp Biochem Physiol B 143:209–218

    Article  CAS  PubMed  Google Scholar 

  • de la Parra AM, Rosas A, Lazo JP, Viana MT (2007) Partial characterization of the digestive enzymes of Pacific bluefin tuna Thunnus orientalis under culture conditions. Fish Physiol Biochem 33:223–231

    Article  CAS  Google Scholar 

  • Erlanger BF, Kokowsky N, Cohen W (1961) The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys 95:271–278

    Article  CAS  PubMed  Google Scholar 

  • Galaviz M, Garcia-Gasca A, Drawbridge M, Alvarez-Gonzalez C, Lopez L (2011) Ontogeny of the digestive tract and enzymatic activity in white seabass, Atractoscion nobilis, larvae. Aquaculture 318:162–168

    Article  CAS  Google Scholar 

  • García-Gasca A, Galaviz M, Gutierréz J, García-Ortega A (2006) Development of the digestive tract, trypsin activity and gene expression in eggs and larvae of the bullseye puffer fish Sphoeroides annulatus. Aquaculture 251:366–376

    Article  Google Scholar 

  • Geiger R (1986) Chymotrypsin. VCH, Weinheim

    Google Scholar 

  • Gjellesvik DR, Lombardo D, Walther BT (1992) Pancreatic bile salt dependent lipase from cod (Gadus morhua): purification and properties. Biochim Biophys Acta 1124:123–134

    Article  CAS  PubMed  Google Scholar 

  • He T, Xiao Z, Liu Q, Ma D, Xu S, Xiao Y, Li J (2012) Ontogeny of the digestive tract and enzymes in rock bream Oplegnathus fasciatus (Temminck et Schlegel 1844) larvae. Fish Physiol Biochem 38:297–308

    Article  CAS  PubMed  Google Scholar 

  • Heu MS, Kim HR, Pyeun JH (1995) Comparison of trypsin and chymotrypsin from the viscera of anchovy, Engraulis japonica. Comp Biochem Physiol B 112:557–567

    Article  CAS  PubMed  Google Scholar 

  • Jimenez-Martinez LD, Alvarez-González CA, Tovar-Ramírez D, Gaxiola G, Sanchez-Zamora A, Moyano FJ, Alarcón FJ, Márquez-Couturier G, Gisbert E, Contreras-Sánchez WM, Perales-García N, Arias-Rodríguez L, Indy JR, Páramo-Delgadillo S, Palomino-Albarrán IG (2012) Digestive enzyme activities during early ontogeny in Common snook (Centropomus undecimalis). Fish Physiol Biochem 38:441–454

    Article  CAS  PubMed  Google Scholar 

  • Kagawa H, Tanaka H, Ohta H, Okuzawa K, Iinuma N (1997) Induced ovulation by injection of 17,20 beta-dihydroxy-4-pregnen-3-one in the artificially matured Japanese eel, with special reference to ovulation time. Fish Sci 63:365–367

    CAS  Google Scholar 

  • Kagawa H, Sakurai Y, Horiuchi R, Kazeto Y, Gen K, Imaizumi H, Masuda Y (2012) Mechanism of oocyte maturation and ovulation and its application to seed production in the Japanese eel. Fish Physiol Biochem. doi:10.1007/s10695-012-9607-3

    PubMed  Google Scholar 

  • Kanno G, Yamaguchi T, Kishimura H, Yamaha E, Saeki H (2010) Purification and characteristics of trypsin from masu salmon (Oncorhynchus masou) cultured in fresh-water. Fish Physiol Biochem 36:637–645

    Article  CAS  PubMed  Google Scholar 

  • Kishimura H, Hayashi K, Miyashita Y, Nonami Y (2006a) Characteristics of trypsins from the viscera of true sardine (Sardinops melanostictus) and the pyloric ceca of arabesque greenling (Pleuroprammus azonus). Food Chem 97:65–70

    Article  CAS  Google Scholar 

  • Kishimura H, Tokuda Y, Klomklao S, Benjakul S, Ando S (2006b) Comparative study of enzymatic characteristics of trypsins from the pyloric ceca of yellow tail (Seriola quinqueradiata) and brown hakeling (Physiculus japonicus). J Food Biochem 30:521–534

    Article  CAS  Google Scholar 

  • Kishimura H, Tokuda Y, Yabe M, Klomklao S, Benjakul S, Ando S (2007) Trypsins from the pyloric ceca of jacopever (Sebastes schlegelii) and elkhorn sculpin (Alcichthys alcicornis): Isolation and characterization. Food Chem 100:1490–1495

    Article  CAS  Google Scholar 

  • Kishimura H, Klomklao S, Benjakul S, Chun B (2008) Characteristics of trypsin from the pyloric ceca of walleye pollock (Theragra chalcogramma). Food Chem 106:194–199

    Article  CAS  Google Scholar 

  • Klomklao S, Benjakul S, Visessanguan W, Kishimura H, Simpson BK (2006a) Purification and characterization of trypsin from the spleen of tongol tuna (Thunnus tonggol). J Agric Food Chem 54:5617–5622

    Article  CAS  PubMed  Google Scholar 

  • Klomklao S, Benjakul S, Visessanguan W, Kishimura H, Simpson BK, Saeki H (2006b) Trypsins from yellowfin tuna (Thunnus albacores) spleen: purification and characterization. Comp Biochem Physiol B 144:47–56

    Article  PubMed  Google Scholar 

  • Klomklao S, Benjakul S, Visessanguan W, Kishimura H, Simpson BK (2007) Trypsin from the pyloric caeca of bluefish (Pomatomus saltatrix). Comp Biochem Physiol B 148:382–389

    Article  PubMed  Google Scholar 

  • Kolkovski S (2001) Digestive enzymes in fish larvae and juveniles—implications and applications to formulated diets. Aquaculture 200:181–201

    Article  CAS  Google Scholar 

  • Kortner TM, Overrein I, Oie G, Kjørsvik E, Bardal T, Wold PA, Arukwe A (2011) Molecular ontogenesis of digestive capability and associated endocrine control in Atlantic cod (Gadus morhua) larvae. Comp Biochem Physiol A 160:190–199

    Article  CAS  Google Scholar 

  • Kristjánsson MM, Nielsen HH (1992) Purification and characterization of two chymotrypsin-like proteases from the pyloric caeca of rainbow trout (Oncorhynchus mykiss). Comp Biochem Physiol B 101:247–253

    PubMed  Google Scholar 

  • Kurokawa T, Kagawa H, Ohta H, Tanaka H, Okuzawa K, Hirose K (1995) Development of digestive organs and feeding ability in larvae of Japanese eel (Anguilla japonica). Can J Fish Aquat Sci 52:1030–1036

    Article  Google Scholar 

  • Kurokawa T, Suzuki T, Ohta H, Kagawa H, Tanaka H, Unuma T (2002) Expression of pancreatic enzyme genes during the early larval stage of Japanese eel Anguilla japonica. Fish Sci 68:736–744

    Article  CAS  Google Scholar 

  • Kurokawa T, Koshio M, Kaiya H, Hashimoto H, Nomura K, Uji S, Awaji M, Gen K, Tanaka H (2011) Distribution of pepsinogen- and ghrelin-producing cells in the digestive tract of Japanese eel (Anguilla japonica) during metamorphosis and the adult stage. Gen Comp Endocrinol 173:475–482

    Article  CAS  PubMed  Google Scholar 

  • Lazo J, Holt G, Arnold C (2000) Ontogeny of pancreatic enzymes in larval red drum Sciaenops ocellatus. Aquac Nutr 6:183–192

    Article  CAS  Google Scholar 

  • Lazo J, Mendoza R, Holt G, Aguilera C, Arnold C (2007) Characterization of digestive enzymes during larval development of red drum (Sciaenops ocellatus). Aquaculture 265:194–205

    Article  CAS  Google Scholar 

  • Liu Z, Wang Z, Xu S, Xu L (2007) Two trypsin isoforms from the intestine of the grass carp (Ctenopharyngodon idellus). J Comp Physiol B 177:655–666

    Article  CAS  PubMed  Google Scholar 

  • Lu B, Zhou L, Cai Q, Hara K, Maeda A, Su W, Cao M (2008) Purification and characterisation of trypsins from the pyloric caeca of mandarin fish (Siniperca chuatsi). Food Chem 110:352–360

    Article  CAS  Google Scholar 

  • Miura C, Ohta T, Ozaki Y, Tanaka H, Miura T (2009) Trypsin is a multifunctional factor in spermatogenesis. Proc Natl Acad Sci USA 106:20972–20977

    Article  CAS  PubMed  Google Scholar 

  • Moyano F, Díaz M, Alarcón F, Sarasquete M (1996) Characterization of digestive enzyme activity during larval development of gilthead seabream (Sparus aurata). Fish Physiol Biochem 15:121–130

    Article  CAS  Google Scholar 

  • Natalia Y, Hashim R, Ali A, Chong A (2004) Characterization of digestive enzymes in a carnivorous ornamental fish, the Asian bony tongue Scleropages formosus (Osteoglossidae). Aquaculture 233:305–320

    Article  CAS  Google Scholar 

  • Nolasco H, Moyano-López F, Vega-Villasante F (2011) Partial characterization of pyloric-duodenal lipase of gilthead seabream (Sparus aurata). Fish Physiol Biochem 37:43–52

    Article  CAS  PubMed  Google Scholar 

  • Ohta H, Kagawa H, Tanaka H, Okuzawa K, Hirose K (1996) Milt production in the Japanese eel Anguilla japonica induced by repeated injections of human chorionic gonadotropin. Fish Sci 62:44–49

    CAS  Google Scholar 

  • Ohta H, Kagawa H, Tanaka H, Okuzawa K, Iinuma N, Hirose K (1997) Artificial induction of maturation and fertilization in the Japanese eel, Anguilla japonica. Fish Physiol Biochem 17:163–169

    Article  CAS  Google Scholar 

  • Oozeki Y, Bailey K (1995) Ontogenetic development of digestive enzyme-activities in larval walleye pollock, Theragra chalcogramma. Mar Biol 122:177–186

    CAS  Google Scholar 

  • Ozaki Y, Tanaka H, Kagawa H, Ohta H, Adachi S, Yamauchi K (2006) Fine structure and differentiation of the alimentary canal in captive-bred Japanese eel Anguilla japonica preleptocephali. Fish Sci 72:13–19

    Article  CAS  Google Scholar 

  • Pedersen B, Ueberschär B, Kurokawa T (2003) Digestive response and rates of growth in pre-leptocephalus larvae of the Japanese eel Anguilla japonica reared on artificial diets. Aquaculture 215:321–338

    Article  Google Scholar 

  • Perez-Casanova J, Murray H, Gallant J, Ross N, Douglas S, Johnson S (2006) Development of the digestive capacity in larvae of haddock (Melanogrammus aeglefinus) and Atlantic cod (Gadus morhua). Aquaculture 251:377–401

    Article  Google Scholar 

  • Raae AJ, Walther BT (1989) Purification and characterization of chymotrypsin, trypsin and elastase like proteinases from cod (Gadus morhua L.). Comp Biochem Physiol B 93:317–324

    CAS  PubMed  Google Scholar 

  • Rønnestad I, Kamisaka Y, Conceição L, Morais S, Tonheim S (2007) Digestive physiology of marine fish larvae: Hormonal control and processing capacity for proteins, peptides and amino acids. Aquaculture 268:82–97

    Article  Google Scholar 

  • Sæle Ø, Nordgreen A, Olsvik PA, Hamre K (2010) Characterization and expression of digestive neutral lipases during ontogeny of Atlantic cod (Gadus morhua). Comp Biochem Physiol A 157:252–259

    Article  Google Scholar 

  • Smichi N, Fendri A, Chaâbouni R, Ben Rebah F, Gargouri Y, Miled N (2010) Purification and biochemical characterization of an acid-stable lipase from the pyloric caeca of sardine (Sardinella aurita). Appl Biochem Biotechnol 162:1483–1496

    Article  CAS  PubMed  Google Scholar 

  • Srivastava AS, Kurokawa T, Suzuki T (2002) mRNA expression of pancreatic enzyme precursors and estimation of protein digestibility in first feeding larvae of the Japanese flounder, Paralichthys olivaceus. Comp Biochem Physiol A 132:629–635

    Article  Google Scholar 

  • Stevens ED, McLeese JM (1984) Why bluefin tuna have warm tummies: temperature effect on trypsin and chymotrypsin. Am J Physiol 246:R487–R494

    CAS  PubMed  Google Scholar 

  • Tanaka H, Kagawa H, Ohta H (2001) Production of leptocephali of Japanese eel (Anguilla japonica) in captivity. Aquaculture 201:51–60

    Article  Google Scholar 

  • Tanaka H, Kagawa H, Ohta H, Unuma T, Nomura K (2003) The first production of glass eel in captivity: fish reproductive physiology facilitates great progress in aquaculture. Fish Physiol Biochem 28:493–497

    Article  Google Scholar 

  • Tsukamoto K, Chow S, Otake T, Kurogi H, Mochioka N, Miller MJ, Aoyama J, Kimura S, Watanabe S, Yoshinaga T, Shinoda A, Kuroki M, Oya M, Watanabe T, Hata K, Ijiri S, Kazeto Y, Nomura K, Tanaka H (2011) Oceanic spawning ecology of freshwater eels in the western North Pacific. Nat Commun 2:179

    Article  PubMed  Google Scholar 

  • Yang F, Su W, Lu B, Wu T, Sun L, Hara K, Cao M (2009) Purification and characterization of chymotrypsins from the hepatopancreas of crucian carp (Carassius auratus). Food Chem 116:860–866

    Article  CAS  Google Scholar 

  • Yoshinaka R, Sato M, Suzuki T, Ikeda S (1985) Purification and some properties of two anionic trypsins from the eel (Anguilla japonica). Comp Biochem Physiol B 80:5–9

    Article  CAS  PubMed  Google Scholar 

  • Zhou LZ, Ruan MM, Cai QF, Liu GM, Sun LC, Su WJ, Cao MJ (2012) Purification, characterization and cDNA cloning of a trypsin from the hepatopancreas of snakehead (Channa argus). Comp Biochem Physiol B 161:247–254

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to Dr. Y. Masuda, Mr. H. Imaizumi, Mr. T. Jinbo (Sibushi Laboratory, NRIA) and Mr. H. Hashimoto (Yaeyama Laboratory, Seikai National Fisheries Research Institute) for providing the Japanese eel larvae. This study was supported by a grant-in-aid for “Development of Sustainable Aquaculture Technology Independent of Wild Fishery Resources” from the Ministry of Agriculture, Forestry and Fisheries, Government of Japan.

Author information

Authors and Affiliations

  1. National Research Institute of Aquaculture, Fisheries Research Agency, 224-1, Hiruta, Tamaki, Mie, 519-0423, Japan

    Koji Murashita, Hirofumi Furuita, Hiroyuki Matsunari, Takeshi Yamamoto, Masahiko Awaji, Kazuharu Nomura, Jiro Nagao & Hideki Tanaka

Authors
  1. Koji Murashita
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hirofumi Furuita
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiroyuki Matsunari
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takeshi Yamamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Masahiko Awaji
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kazuharu Nomura
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiro Nagao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hideki Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Koji Murashita.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Murashita, K., Furuita, H., Matsunari, H. et al. Partial characterization and ontogenetic development of pancreatic digestive enzymes in Japanese eel Anguilla japonica larvae. Fish Physiol Biochem 39, 895–905 (2013). https://doi.org/10.1007/s10695-012-9749-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10695-012-9749-3

Keywords

Search

Navigation