Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Isolation and immunological characterization of three highly purified serine proteinases from Antarctic krill (Euphausia superba)


Three individual serine proteinases (I, II, III) originating from Antarctic krill (E. superba) were separated and highly purified using a combination of affinity and high resolution ion exchange chromatography. Each enzyme showed a single protein band (30 000 Daltons) in sodium dodecyl sulphate polyacrylamide gradient gel electrophoresis indicating high purity and identical molecular weights. Moreover, each enzyme demonstrated one immunoprecipitate on crossed immunoelectrophoresis (two-dimensional agarose gel electrophoresis) using polyclonal rabbit antibodies which confirmed the high purity of the individual enzymes. A mixture of the three enzymes (I, II, III) revealed two immunoprecipitates, not one or three which should have been the case for identical or non-identical immunological properties. Double immunodiffusion test according to Ouchterlony exhibited immunological identity between enzyme II and III. Enzyme I showed only partial identity with II/III. These findings correlated well with biochemical data on the three serine proteinases. Enzyme I is able to liberate free amino acids from polypeptides in comparison with enzyme II and III (classical true endopeptidases), which do not. We suggest that these unique biochemical properties also have their immunological counterpart expressed as other antigenic determinants of the molecular structure.

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


  1. Axelsen NH, Krøll J, Weeke B (1973) A manual of quantitative immunoelectrophoresis: Methods and applications. Scand J Immunol [Suppl 1] 2

  2. Bucht A, Karlstam B (1986) Immunological characterization of three highly purified trypsin-like enzymes from Antarctic krill (Euphausia superba). Biol Chem Hoppe-Seyler 367:366

  3. Folk JE, Schirmer EW (1963) The porcine pancreatic carboxypeptidase A system. J Biol Chem 238:3884–3894

  4. Folk JE, Piez KA, Carroll WR, Gladner JA (1960) Carboxypeptidase B: purification and characterization of the porcine enzyme. J Biol Chem 235:2272–2277

  5. Harboe N, Ingild A (1973) Immunization, isolation of immunoglobulins, estimation of antibody titre. Scand J Immunol [Suppl 1] 2:161–164

  6. Kimoto K, Kusuma S, Murakami K (1983) Purification and characterization of serine proteinases from Euphausia superba. Agric Biol Chem 47:529–534

  7. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

  8. Norrman A, Karlstam B (1985) Immunochemical characterization of proteins in Antarctic krill (Euphausia superba). Abstract and Poster at 13th Int Congr Biochem, Amsterdam, p 94

  9. Osnes KK, Mohr V (1985) On the purification and characterization of three anionic serine-type peptide hydrolases from Antarctic krill, Euphausia superba. Comp Biochem Physiol 82B:607–619

  10. Rick W (1974) In: Bergmeyer HU (eds) Methods of enzymatic analysis, vol 2. Academic Press, New York, pp 1013–1023

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Bucht, A., Karlstam, B. Isolation and immunological characterization of three highly purified serine proteinases from Antarctic krill (Euphausia superba). Polar Biol 11, 495–500 (1991).

Download citation


  • Serine Proteinase
  • Free Amino Acid
  • Sodium Dodecyl Sulphate
  • Endopeptidase
  • Antarctic Krill