Enzyme-assisted extraction of anticoagulant polysaccharide from Liparis tessellatus eggs.

Autor: Ticar BF; Department of Seafood Science and Technology, The Institute of Marine Industry, Gyeongsang National University, Tongyeong 650-160, Republic of Korea; Western Visayas College of Science and Technology, College of Arts and Sciences, Burgos St., La Paz, Iloilo City, Iloilo, Philippines. Electronic address: berna2423@gmail.com., Rohmah Z; Department of Seafood Science and Technology, The Institute of Marine Industry, Gyeongsang National University, Tongyeong 650-160, Republic of Korea; Faculty of Biology, Universitas Gadjah Mada, Yogyakarta 55281, Indonesia., Ambut CV; Western Visayas College of Science and Technology, College of Arts and Sciences, Burgos St., La Paz, Iloilo City, Iloilo, Philippines., Choi YJ; Department of Seafood Science and Technology, The Institute of Marine Industry, Gyeongsang National University, Tongyeong 650-160, Republic of Korea., Mussatto SI; Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC Delft, The Netherlands., Choi BD; Department of Seafood Science and Technology, The Institute of Marine Industry, Gyeongsang National University, Tongyeong 650-160, Republic of Korea. Electronic address: bdchoi@gnu.ac.kr.
Jazyk: angličtina
Zdroj: International journal of biological macromolecules [Int J Biol Macromol] 2015 Mar; Vol. 74, pp. 601-7. Date of Electronic Publication: 2015 Jan 09.
DOI: 10.1016/j.ijbiomac.2015.01.002
Abstrakt: This study aimed to recover a heparin-like anticoagulant polysaccharide from Liparis tessellatus eggs (PLE) by using enzyme-assisted extraction technique. Extraction experiments were carried out using three different enzymes (Alcalase®2.4 L, Flavourzyme®500 MG, and Protamex®) under different conditions of temperature (45, 50, and 55°C), pH (6.5, 7.0, and 7.5), incubation time (24, 36, and 48 h), and enzyme to substrate ratio (E/S=0.5, 1.0, and 1.5%, w/w), which were combined according to a D-optimal design. Statistical analysis of extraction results allowed identifying the variables with greater influence on the extraction yield, and selecting the conditions that maximize the PLE extraction. The best extraction results were achieved when using the Protamex® enzyme in an E/S ratio of 1.34% (w/w), pH 6.60, 47.40°C, during 26.50 h. Under these conditions, a polysaccharide yield of 2.10% (w/w) was obtained. Clotting time measurements, activated partial thromboplastin time, and prothrombin time for evaluation of the anticoagulant properties of PLE were determined and showed increasing activities in correlation with the concentrations used. In the final step, the heparin-like nature of PLE was confirmed by digestion with heparinases I, II, and III, which showed ΔDiHS-0S, ΔDiHS-6S, ΔDiHS-diS1, and ΔDiHS-diS2 at compositions of 0.04, 0.03, 0.35, and 0.24 mol/g, respectively.
(Copyright © 2015 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: