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Purification of α‐amylase from human saliva by superparamagnetic particles
Author(s) -
Lin Jenshinn,
Lin YeongShenn,
Kuo ShoTin,
Jiang ChiiMing,
Wu MingChang
Publication year - 2009
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.3450
Subject(s) - amylase , starch , epichlorohydrin , chromatography , chemistry , superparamagnetism , saliva , polyacrylamide , gel electrophoresis , polyacrylamide gel electrophoresis , sodium dodecyl sulfate , enzyme , biochemistry , organic chemistry , polymer chemistry , magnetic field , physics , magnetization , quantum mechanics
BACKGROUND: Several studies of magnetic carrier technology have focused on the application of separation technology, because the magnetic support can separate the target from the reaction medium by application of a magnetic field and because the magnetic carrier can be easily recovered. In the present study, superparamagnetic iron oxide (SPIO) modified by epichlorohydrin was employed as a support whose surface could be coated with starch. The starch‐coated support was used for isolating human salivary amylases. RESULTS: The results showed that the starch‐SPIO support could isolate amylases from human saliva with 91.1% recovery and 3.5‐fold purification to high specific activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the purifed amylase comprised two isoamylases with estimated molecular weights of 55 and 59 kDa. The activities of crude and purified amylases showed optimal pH values of 6–7 and 7 and optimal temperatures of 40 and 30 °C respectively. The thermal stability range for both crude and purified amylases was between 20 and 40 °C. CONCLUSION: The attachment of substrates to SPIO could offer a novel and efficient method for purifying enzymes either as an initial step prior to further purification or as a final step for diagnostic usage. Copyright © 2009 Society of Chemical Industry