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KMID : 1094720070120050542
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Biotechnology and Bioprocess Engineering
2007 Volume.12 No. 5 p.542 ~ p.547
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Solubilization conditions for hydrophobic membrane protein, oleosin, in soybeans
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Kim Hyun
Kim So-Young
Han Nam-Soo
Tao Bernard Y.
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Abstract
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Oleosins are amphipathic proteins found in soybeans that give stability to oil-bodies by surrounding them. Furthermore, oleosins have broad biotechnological applications in protein purification and in immobilization as a tagging parther. In this study we provide the proper combinations of detergents and chaotropes for solubilization of oleosins. The solubilization effects of each detergent were compared by the results of SDS-PAGE analysis, from the density measurement of each band using an image analysis program. Among the twelve chemicals tested, 3-(decyldimethylammonio) propanesulfonale inner salt (SB 3?10), nonaethylene glycol octylphenyl ether (NP-40), and n-dodecyl-β-d-maltoside (DM) were determined to be effective for oleosin solubilization. In addition, urea, thiourea, and reducing agents played roles in enhancing the effects of the detergents. The best results were obtained with 2.5% of each detergent in combination with 4 M urea and 2 M thiourea at room temperature. The zwitterionic detergent, SB 3?10, is recommended as the best non-denaturant for the solubilization of soybean oleosins with the advantage of its easy removal by dialysis due to its low molecular weight and a high critical micelle concentration.
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KEYWORD
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hydrophobic protein, solubilization, detergents, SDS-PAGE, oleosin
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