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Cold-adapted structural properties of trypsins from walleye pollock (Theragra chalcogramma) and Arctic cod (Boreogadus saida)

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Abstract

Complementary DNA clones encoding trypsins were isolated from pyloric ceca of cold-adapted fish, walleye pollock (Theragra chalcogramma) (WP-T) and Arctic cod (Boreogadus saida) (AC-T). The isolated full-length cDNA clones of WP-T and AC-T were 852 and 860 bp, respectively, and both cDNAs were contained an open reading frame of 726 bp. WP-T and AC-T seemed to be synthesized as preproenzyme that contains a signal peptide, an activation peptide, and a mature trypsin. Although the amino acid sequence identities of WP-T and AC-T to that of bovine trypsin were 64 and 63%, respectively, they completely conserved the structural features for catalytic function of trypsin. On the other hand, WP-T and AC-T possessed the four Met residues (Met135, Met145, Met175 and Met242) in their molecules and the deletion of Tyr151 and substitution of Pro152 for Gly in their autolysis loops when aligned with the sequences of tropical-zone fish and bovine trypsins. In addition, the contents of charged amino acid residues at the N-terminal regions (positions 20–50) of WP-T and AC-T were extremely higher than those of other fish and bovine trypsins. Moreover, one amino acid (Asn72) and two amino acids (Asn72 and Val75) coordinating with Ca2+ in bovine trypsin were exchanged for another amino acids in WP-T (His) and AC-T (His and Glu), respectively, and the contents of negative charged amino acids at their Ca2+-binding regions were lower than those of tropical-zone fish and bovine trypsins. Therefore, it was considered that these structural characteristics of WP-T and AC-T are closely related to their lower thermostability.

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Acknowledgments

We would like to thank crew and officers of the T/S Oshoro Maru, Hokkaido University, for in collecting Arctic cod. This research was partly supported by the grant from HOKUSUI Association.

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Authors and Affiliations

  1. Laboratory of Marine Products and Food Science, Research Faculty of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, 041-8611, Japan

    Gaku Kanno, Hideki Kishimura & Hiroki Saeki

  2. Field Science Center for Northern Biosphere, Hokkaido University, Hakodate, Hokkaido, 041-8611, Japan

    Jun Yamamoto

  3. Department of Natural Sciences, Faculty of Health and Welfare Science, Nayoro City University, Nayoro, Hokkaido, 096-8641, Japan

    Seiichi Ando

  4. Research and Development Department, Hokkaido Industrial Technology Center, Hakodate, Hokkaido, 041-0801, Japan

    Takeshi Shimizu

  5. Department of Food Technology, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand

    Soottawat Benjakul

  6. Department of Food Science and Technology, Faculty of Technology and Community Development, Thaksin University, Phattalung Campus, Phattalung, 93110, Thailand

    Sappasith Klomklao

  7. Faculty of Agro-Industry, King Mongkut’s Institute of Technology Ladkrabang, Choakhunthaharn Building, Choakhunthaharn Rd., Ladkrabang, Bangkok, 10520, Thailand

    Sitthipong Nalinanon

  8. Faculty of Food Science and Biotechnology, Pukyong National University, Busan, 608-737, Republic of Korea

    Byung-Soo Chun

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  1. Gaku Kanno
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  2. Hideki Kishimura
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  9. Byung-Soo Chun
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  10. Hiroki Saeki
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Correspondence to Hideki Kishimura.

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Kanno, G., Kishimura, H., Yamamoto, J. et al. Cold-adapted structural properties of trypsins from walleye pollock (Theragra chalcogramma) and Arctic cod (Boreogadus saida). Eur Food Res Technol 233, 963–972 (2011). https://doi.org/10.1007/s00217-011-1592-8

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  • DOI: https://doi.org/10.1007/s00217-011-1592-8

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