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Novel Chymotrypsin Purified and Biochemically Characterized from Digestive Organs of Bigfin Reef Squid ( Sepioteuthis lessoniana )

Author

Listed:
  • Jirapan Satjarak

    (Aquatic Science and Innovative Management Division, Faculty of Natural Resources, Prince of Songkla University, Songkhla 90112, Thailand)

  • Sappasith Klomklao

    (Department of Food Science and Technology, Faculty of Agro and Bio Industry, Thaksin University, Phattalung Campus, Phattalung 93110, Thailand)

  • Yi Zhang

    (Department of Food Science, College of Agricultural Sciences, The Pennsylvania State University, University Park, PA 16802, USA)

  • Karun Thongprajukaew

    (Division of Health and Applied Sciences, Faculty of Science, Prince of Songkla University, Songkhla 90112, Thailand)

Abstract

Chymotrypsin from the digestive organs of bigfin reef squid, a major commercial squid species in Thailand, was purified to 41-fold with 5.7% yield by a sequential purification process including ammonium sulfate precipitation, size exclusion, and ion exchange chromatography. According to SDS-PAGE, the molecular weight of purified chymotrypsin was 43 kDa. Native-PAGE analysis revealed a single band for this purified enzyme. The optimum pH and temperature for chymotrypsin activity of the purified enzyme were a pH of 7.0 with a temperature of 55 °C. The purified chymotrypsin remained stable throughout a wide range of pH levels (6–11) and at relatively high temperature (55 °C). It was significantly inhibited by PMSF and TPCK. The values of the kinetic constants K m and K cat were found to be 1.33 mM and 31.46 s −1 , respectively. The purified chymotrypsin has the N -terminal amino acid IVGGQEATPGEWPWQAALQV. This study provided new information about the biochemical properties of pure chymotrypsin from bigfin reef squid, which will be useful in the future investigation, aquaculture, and application of bigfin reef squid.

Suggested Citation

  • Jirapan Satjarak & Sappasith Klomklao & Yi Zhang & Karun Thongprajukaew, 2024. "Novel Chymotrypsin Purified and Biochemically Characterized from Digestive Organs of Bigfin Reef Squid ( Sepioteuthis lessoniana )," Resources, MDPI, vol. 13(5), pages 1-12, May.
    • Handle: RePEc:gam:jresou:v:13:y:2024:i:5:p:67-:d:1394434
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