极地研究

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Methodological approach to the isolation of functionally active proteins from the tissues of marine hydrobionts: an example of Adamussium colbecki

Nataliia Raksha*, Dmytro Gladun, Oleksii Savchuk & Lydmila Ostapchenko   

  1. Educational and Scientific Center ‘Institute of Biology’, Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine
  • 出版日期:1965-12-30 发布日期:1965-12-30
  • 通讯作者: Nataliia Raksha

Methodological approach to the isolation of functionally active proteins from the tissues of marine hydrobionts: an example of Adamussium colbecki

Nataliia Raksha*, Dmytro Gladun, Oleksii Savchuk & Lydmila Ostapchenko   

  1. Educational and Scientific Center ‘Institute of Biology’, Taras Shevchenko National University of Kyiv, 64/13, Volodymyrska Str., Kyiv 01601, Ukraine
  • Online:1965-12-30 Published:1965-12-30
  • Contact: Nataliia Raksha

摘要: Enzymes from cold-adapted organisms have significant application potential. Because of their unique properties they have been found to be useful in various industries. Despite indisputable practical interest, cold active enzymes also represent a valuable model for fundamental research into protein folding and catalysis. Many investigators have focused their attention on marine hydrobionts, which are growing in importance as a promising source of enzymes. The nature of the source not only determines the availability and the cost of biomolecules of interest but also determines the choice of method for their extraction.
A simple and convenient methodological approach of two-stage extraction of proteins has been tested on the Antarctic marine hydrobiont—Adamussium colbecki. This method extracts enough effective protein directly from primary raw materials, as well as when using leftover crude precipitates. The electrophoretic pattern of proteins showed the presence of molecules in a wide range of molecular weights in the samples of A. colbecki after the first and the second stage of extraction. The general proteolytic activity in the first and the second extracts were examined using a zymogram technique. Our experiments revealed that the second extract of A. colbecki contained thermo stable protease exhibiting a molecular weight of 95 kDa in a gelatin zymogram. Further biochemical assays, using different substrates, were conducted to partially identify the types of hydrolases present in the first and the second extracts. Our results revealed the presence of enzymes with collagenolytic and some amylolytic activities preserved in the second extracts. But no esterase or amidase trypsin-like activities were found in the second extract, in contrast to the first extract where this type of activity was significant.

关键词: Antarctic marine hydrobiont, protein extraction, enzyme activity determination

Abstract: Enzymes from cold-adapted organisms have significant application potential. Because of their unique properties they have been found to be useful in various industries. Despite indisputable practical interest, cold active enzymes also represent a valuable model for fundamental research into protein folding and catalysis. Many investigators have focused their attention on marine hydrobionts, which are growing in importance as a promising source of enzymes. The nature of the source not only determines the availability and the cost of biomolecules of interest but also determines the choice of method for their extraction.
A simple and convenient methodological approach of two-stage extraction of proteins has been tested on the Antarctic marine hydrobiont—Adamussium colbecki. This method extracts enough effective protein directly from primary raw materials, as well as when using leftover crude precipitates. The electrophoretic pattern of proteins showed the presence of molecules in a wide range of molecular weights in the samples of A. colbecki after the first and the second stage of extraction. The general proteolytic activity in the first and the second extracts were examined using a zymogram technique. Our experiments revealed that the second extract of A. colbecki contained thermo stable protease exhibiting a molecular weight of 95 kDa in a gelatin zymogram. Further biochemical assays, using different substrates, were conducted to partially identify the types of hydrolases present in the first and the second extracts. Our results revealed the presence of enzymes with collagenolytic and some amylolytic activities preserved in the second extracts. But no esterase or amidase trypsin-like activities were found in the second extract, in contrast to the first extract where this type of activity was significant.

Key words: Antarctic marine hydrobiont, protein extraction, enzyme activity determination