Salt‐induced phase separation of CHAPS and potential application for hydrophobic molecule extraction
Salt‐induced phase separation of CHAPS and potential application for hydrophobic molecule extraction
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CHAPS 的盐诱导相分离及其在疏水性分子萃取中的潜在应用
DOI：
10.1002/jctb.6016
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发表时间：
2019
影响因子：
3.4
通讯作者：
Liew Mervyn WO
中科院分区：
文献类型：
--
作者：
Lian Ai A;Tan Soo C;Nagamine Toshihiro;Liew Mervyn WO
背景3‐[（3‐胆酰胺丙基）二甲酰胺]‐1‐丙磺酸盐（CHAPS）是一种两性离子洗涤剂，通常用于基于蛋白质生物化学的应用。本研究首次证明了通过盐的加入，CHAPS的相分离是成功的。固溶液温度行为的阐明以及双节点曲线和系带线长度（TLL）的测定使基于CHAPS的相分离系统的后续开发成为可能。然后使用低分子量染料考马斯亮蓝（CBB）和整体膜蛋白细胞色素P450 (CYP) 2C19对系统进行评估。结果发现盐的阴离子特性至关重要，硫酸盐和磷酸盐在4°C下成功诱导CHAPS相分离。后来发现CHAPS和（NH4） 2so4混合物具有较高的溶质温度（UCST）。收集双节点数据并拟合到一个方程中，以确定形成相分离系统所需的CHAPS和（NH4） 2so4的最小量。在CHAPS/(NH4) 2so4体系中，CBB被广泛分配到富胶束相（MRP），提取率为83±0.32%，而CYP 2C19被成功回收，提取率为46±7.58%。结论低温下硫酸盐加成的CHAPS相分离可用于疏水分子的提取。本研究结果提出了CHAPS作为相分离系统组成部分的应用范围。©2019化学工业协会
BACKGROUND3‐[(3‐Cholamidopropyl)dimethylammonio]‐1‐propanesulfonate (CHAPS) is a zwitterionic detergent commonly utilized in protein biochemistry‐based applications. This study demonstrates for the first time the successful phase separation of CHAPS via salt addition. Elucidation of consolute solution temperature behavior and determination of binodal curve and tie‐line length (TLL) enabled the subsequent development of CHAPS‐based phase separation systems. The systems were then evaluated using a low‐molecular‐weight dye, Coomassie Brilliant Blue (CBB), and integral membrane protein, cytochrome P450 (CYP) 2C19.RESULTSThe anionic identity of salts was found to be critical, with sulfate and phosphate salts being successful in inducing phase separation of CHAPS at 4 °C. CHAPS and (NH4)2SO4mixtures were later found to have an upper consolute solution temperature (UCST). Binodal data were collected and fitted to an equation to allow determination of the minimum amount of CHAPS and (NH4)2SO4required for the formation of phase separation systems. In CHAPS/(NH4)2SO4systems, CBB was found to partition extensively to the micelle‐rich phase (MRP) with an extraction yield of 83 ± 0.32%, while CYP 2C19 was successfully recovered with an extraction yield of 46 ± 7.58%.CONCLUSIONPhase separation of CHAPS achieved at low temperature via sulfate salt addition can be exploited for the extraction of hydrophobic molecules. The outcomes of this study propound the scope for application of CHAPS as a component of phase separation systems. © 2019 Society of Chemical Industry