Skip to main content
SpringerLink
Log in

Partial characterization of the digestive enzymes of Pacific bluefin tuna Thunnus orientalis under culture conditions

Fish Physiology and Biochemistry volume 33, pages 223–231 (2007)Cite this article

Abstract

The digestive enzyme activities of Pacific bluefin tuna Thunnus orientalis were evaluated for specific activity and characterized for pH and temperature optima in crude extracts of stomach, caecal mass, and proximal, middle and distal intestine. A higher level of alkaline proteolytic activity was detected in the caecal mass than in the proximal intestine. Total alkaline proteases, trypsin, chymotrypsin and leucine aminopeptidase (LAP) were tested. The temperature and pH analyses showed that proteolytic activity as well as lipase were maximal in the alkaline range, with a maximum at pH 9.0 and at temperatures between 35 and 60°C, except for the pepsin, which showed maximum activity at the same temperatures but in the acid range (pH 3.0). The α-amylase activity showed a broader range in activity, both for pH and temperature, with higher activity over the alkaline pH values and higher temperature. The lipase activity seems to be nondependent on bile salts under our assay conditions, resulting in a significant activity reduction in the presence of bile salts. This knowledge will allow the development of a gastrointestinal model (everted intestine) where food or feed will be hydrolysed with the fish’s own enzymes, a project that is being undertaken in our laboratory as a contribution to the development of novel diets for tuna fish.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price includes VAT (Canada)

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  • Appel W (1974) Aminopeptidase and amino acid acrylamidase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic, New York, pp 950–978

    Google Scholar 

  • Bernfeld P (1955) Amylase α and β: colorimetric assay method. In: Colowich SP, Kaplan NO (eds) Methods in enzymology. Academic, New York, pp 149–157

    Chapter  Google Scholar 

  • Borgström B, Erlanson CH (1973) Pancreatic lipase and co-lipase: interactions and effects of bile salts and other detergents. Eur J Biochem 37:60–68

    Article  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Castillo-Yáñez FJ, Pacheco-Aguilar R, Garcia-Carreño FL, Navarrete-Del Toro MA (2005) Isolation and characterization of trypsin from pyloric caeca of Monterey sardine Sardinops sagax caerulea. Comp Biochem Physiol 140B:91–98

    Google Scholar 

  • Chakrabarti I, Gani MdA, Chaki KK, Sur R, Misra KK (1995) Digestive enzymes in 11 freshwater teleost fish species in relation to food habit and niche segregation. Comp Biochem Physiol 112A(1):167–177

    Article  CAS  Google Scholar 

  • Clark J, MacDonald NL, Stark JR (1987) Leucine aminopeptidase in the digestive tract of Dover sole (Solea solea L.). Aquaculture 61:231–239

    Article  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Fernandez I, Moyano FJ, Diaz M, Martinez T (2001) Characterization of α-amylase activity in five species of Mediterranean sparid fishes (Sparidae, Teleostei). J Exp Mar Biol Ecol 262:1–12

    Article  CAS  Google Scholar 

  • Gildberg A (1988) Aspartic proteinases in fish and aquatic invertebrates. Comp Biochem Physiol 91B:425–435

    CAS  Google Scholar 

  • Gildberg A, Raa J (1983) Purification and characterization of pepsins from the Arctic fish capelin (Mallotus villosus). Comp Biochem Physiol 75A(3):337–342

    Article  CAS  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

    PubMed  CAS  Google Scholar 

  • Harpaz S, Uni Z (1999) Activity of intestinal mucosal brush border membrane enzymes in relation to the feeding habits of three aquaculture fish species. Comp Biochem Physiol 124A:155–160

    CAS  Google Scholar 

  • Hidalgo MC, Urea E, Sanz A (1999) Comparative study of digestive enzymes in fish with different nutritional habits: proteolytic and amylase activities. Aquaculture 170:267–283

    Article  CAS  Google Scholar 

  • Hummel BCW (1959) A modified spectrophotometric determination of chymotrypsin, trypsin and thrombin. Can J Biochem Physiol 37:1393–1399

    PubMed  CAS  Google Scholar 

  • Kitagawa T, Kimura S, Nakata H, Yamada H (2006) Thermal adaptation of Pacific bluefin tuna Thunnus orientalis to temperate waters. Fish Sci 72:149–156

    Article  CAS  Google Scholar 

  • Kishimura H, Hayashi K, Miyashita Y, Yosiyuki N (2005) Characteristics of two trypsin isozymes from the viscera of Japanese anchovy (Engraulis japonica). J Food Biochem 29:459–469

    Article  CAS  Google Scholar 

  • Kishimura H, Tokuda Y, Klomklao S, Benjakul S, Ando S (2006) 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 

  • Klomklao S, Benjakul S, Visessanguan W (2004) Comparative studies on proteolytic activity of splenic extract from three tuna species commonly used in Thailand. J Food Biochem 28:355–372

    Article  Google Scholar 

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

    CAS  Google Scholar 

  • Klomklao S, Benjakul S, Visessanguan W, Kishimura H, Simpson BK (2007) Purification and characterization of trypsins from the spleen of skipjack tuna (Katsuwonus pelamis). Food Chem 100:1580–1589

    Article  CAS  Google Scholar 

  • Krogdahl A, Bakke-McKellep A (2005) Fasting and refeeding cause rapid changes in intestinal tissue mass and digestive enzyme capacities of Atlantic salmon (Salmo salar L.). Comp Biochem Physiol 141A:450–460

    CAS  Google Scholar 

  • Kuz’mina VV (1985) Distribution of carbohydrase activity along the intestine of several species of fresh water fish. J Ichthyol 25(4):137–143

    Google Scholar 

  • Kuz’mina V, Glatman L, Drabkin V, Gelman A (2003) Amylolytic activity in fish intestinal mucosa: temperature effects. Comp Biochem Physiol 134B:529–534

    CAS  Google Scholar 

  • Liddle RA (1997) Cholecystokinin cells. Annu Rev Physiol 59:221–242

    Article  PubMed  CAS  Google Scholar 

  • Lie Ø, Lambertsen G (1985) Digestive enzymes in cod (Gadus morhua): fatty acid specificity. Comp Biochem Physiol 80B:447–450

    CAS  Google Scholar 

  • Lundstedt LM, Bibiano JF, Moraes G (2004) Digestive enzymes and metabolic profile of Pseudoplatystoma corruscans (Teleostei: Siluriformes) in response to diet composition. Comp Biochem Physiol 137B:331–339

    CAS  Google Scholar 

  • Martinez A, Serra JL (1989) Proteolytic activities in the digestive tract of anchovy (Engraulis encrasicholus). Comp Biochem Physiol 93B:61–66

    CAS  Google Scholar 

  • Morgan RG, Hoffman NE (1971) The interaction of lipase, lipase cofactor and bile salts in triglyceride hydrolysis. Biochim Biophys Acta 248(1):143–148

    PubMed  CAS  Google Scholar 

  • Munilla-Morán R, Saborido-Rey F (1996) Digestive enzymes in marine species. II. Amylase activities in gut from seabream (Sparus aurata), turbot (Scophthalmus maximus) and redfish (Sebastes mentella). Comp Biochem Physiol 113B(4):827–834

    Google Scholar 

  • Patton JS, Warner TG, Benson AA (1977) Partial characterization of the bile salt-dependent triacylcglycerol lipase from the leopard shark pancreas. Biochim Biophys Acta 486:322–330

    PubMed  CAS  Google Scholar 

  • Rust MB (2002) Nutritional physiology. In: Halver JE, Hardy R (eds) Fish nutrition. Academic, New York, pp 367–452

    Google Scholar 

  • Sarath G, De la Motte RS, Wagner FW (1989) Protease assay methods. In: Beynon R, Bond J (eds) Proteolytic enzymes: a practical approach. IRL, Oxford, pp 25–56

    Google Scholar 

  • SigmaStat (2003) Systat® Inc. software, Richmond, California, USA

    Google Scholar 

  • Simpson BK, Haard NF (1984) Purification and characterization of trypsin from the Greenland cod Gadus ogac. I. Kinetic and thermodynamic characteristics. Can J Biochem Cell Biol 62:894–900

    Article  CAS  Google Scholar 

  • Stevens ED, McLeese JM (1984) Why bluefin tuna have warm tummies: temperature effect on trypsin and chymotrypsin. Am J Physiol 246:486–494

    Google Scholar 

  • Stevens ED, Kanwisher JW, Carey FG (2000) Muscle temperature in free-swimming giant Atlantic bluefin tuna (Thunnus thynnus L.). J Thermal Biol 25:419–423

    Article  Google Scholar 

  • Tanji M, Kageyama T, Takahashi K (1988) Tuna pepsinogens and pepsins. Eur J Biochem 177:251–259

    Article  PubMed  CAS  Google Scholar 

  • Tičina V, Grubišić L, Katavić I (2004) Sampling and tagging of live bluefin tuna in growth-out floating cages. Aquaculture Res 35:307–310

    Article  Google Scholar 

  • Uys W, Hecht T (1987) Assays on the digestive enzymes of sharptooth catfish, Clarias gariepinus (Pisces: Clariidae). Aquaculture 63:301–313

    Article  CAS  Google Scholar 

  • Wang HY, Wang YJ, Wang QY, Xue CH, Sun M (2006) Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.). Fish Physiol Biochem 32:179–188

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grant SEP-2004-CO1–45785. The authors thank Emyr Peña for his technical assistance. Grateful acknowledgement is also made to Maricultura del Norte, S.A. de C.V. for donations of personnel and practical local support to obtain samples. We thank Nancy Boston for her editorial work.

Author information

Authors and Affiliations

  1. Departamento de Ecología y Biología Animal, Facultad de Ciencias, Universidad de Vigo, 36310, Vigo, Pontevedra, Spain

    Ana Matus de la Parra

  2. Programa de Maestría y Doctorado en Oceanografía Costera, Facultad de Ciencias Marinas, Universidad Autónoma de Baja California (UABC), Ensenada, BC, Mexico

    Antonio Rosas

  3. Centro de Investigaciones Científicas de Educación Superior de Ensenada (CICESE), Km 107 carretera Tij-Eda, 22860 , Ensenada, BC, Mexico

    Juan Pablo Lazo

  4. Instituto de Investigaciones Oceanológicas, Universidad Autónoma de Baja California, Km 107 carretera Tij-Eda, 22860, Ensenada, BC, Mexico

    Maria Teresa Viana

Authors
  1. Ana Matus de la Parra
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Antonio Rosas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Juan Pablo Lazo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maria Teresa Viana
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Maria Teresa Viana.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

de la Parra, A.M., Rosas, A., Lazo, J.P. et al. Partial characterization of the digestive enzymes of Pacific bluefin tuna Thunnus orientalis under culture conditions. Fish Physiol Biochem 33, 223–231 (2007). https://doi.org/10.1007/s10695-007-9134-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10695-007-9134-9

Keywords

  • Bile salts
  • Digestive enzymes
  • Pacific bluefin tuna
  • Pancreatic lipase
  • Thunnus orientalis

Search

Navigation