EXTRACTION AND CHARACTERIZATION OF SEA CUCUMBER Isostichopus fuscus PROTEASES, COLLECTED AT THE GULF OF CALIFORNIA, MEXICO

  • A.C. Hernandez-Sámano Universidad Autónoma Metropolitana (Iztapalapa).
Keywords: sea cucumber, Isostichopus fuscus, ventral muscle, proteases, enzyme characterization

Abstract

Enzyme extracts obtained from I. fuscus ventral muscle were studied. Peak activity was observed at pH 2, 6, and 8, and 50-60°C, whereas maximum enzymatic stability occurred at pH 2 to 6 (except pH 3) and 0 to 40°C. More than 74% residual activity was retained after heating at 70°C and 80°C for 1 h, confirming the presence of heat-resistant enzymes. The extracts also retained 59% initial activity after 21-day storage at 4-5°C. Hg2+ increased the proteolytic activity in 16%, whereas Cu2+ and Mn2+ caused partial inhibition, suggesting the presence of cysteine- and metallo- proteases. Extracts were inhibited by pepstatin A, β-mercaptoethanol and EDTA, confirming the predominance of aspartyl-, cystein-, and metallo-, proteases. Four proteins were identified by SDS-PAGE (180.6, 114.5, 91.7 and 52.9 kDa). Zymograms on casein confirmed the presence of proteolytic enzymes. After purification by anion exchange chromatography a 49 kDa protease, possible metalloprotease, was detected. Mass spectrometry of the partially purified protein showed homology with trypsin and chain E, leech-derived tryptase inhibitor trypsin complex (LDTI). Due that no homology was found with any previously described metalloproteases, the presence of a novel enzyme is suggested.

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Published
2020-01-17
How to Cite
Hernandez-Sámano, A. (2020). EXTRACTION AND CHARACTERIZATION OF SEA CUCUMBER Isostichopus fuscus PROTEASES, COLLECTED AT THE GULF OF CALIFORNIA, MEXICO. Revista Mexicana De Ingeniería Química, 14(1), 35-47. Retrieved from http://www.rmiq.org/ojs311/index.php/rmiq/article/view/1181
Section
Biotechnology