Protein Selectivity Controlled by Polymer Charge Density and Protein Yield: Carboxylated Polysaccharides versus Sulfated Polysaccharides

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• 作者：Xingfei Li ; Yufei Hua ; Yeming Chen ; Xiangzhen Kong ; Caimeng Zhang
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2016
• 出版时间：November 30, 2016
• 年：2016
• 卷：64
• 期：47
• 页码：9054-9062
• 全文大小：516K
• ISSN：1520-5118

文摘

The effect of polymer charge density on protein selectivity in the presence of carboxylated polysaccharides (CPS) and sulfated polysaccharides (SPS) was investigated for Kunitz trypsin inhibitor/Bowman–Birk protease inhibitor (KTI/BBI, KBM). To determine the conditions for coacervation or precipitation as a function of polymer charge densities, turbidimetric titrations and Tricine–SDS-PAGE were used. Polymer charge density as well as chain flexibility greatly influenced the strength of interactions and protein recovery. Although charge compensation must occur for CPS–KBM complexes, SPS–KBM systems did not require conservation of charge neutrality. Despite their similar isoelectric points, KTI bound preferentially to CPS and SPS due to its higher affinity compared to BBI. Complexation of KBM with the polysaccharide with the lowest charge density, arabic gum, expectedly cannot realize the purification of BBI under conditions where binding to more highly charged polysaccharides occurs. This work will be beneficial to selective purification of target proteins through control of protein–polysaccharide complexation.