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首页> 外文期刊>Biotechnology Progress >Mixed Reverse Micelles Facilitated Downstream Processing of Lipase Involving Water-Oil-Water Liquid Emulsion Membrane
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Mixed Reverse Micelles Facilitated Downstream Processing of Lipase Involving Water-Oil-Water Liquid Emulsion Membrane

机译：混合反向胶束促进脂肪酶下游加工涉及水-油-水液体乳化膜

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摘要

Our earlier work for the first time demonstrated that liquid emulsion membrane (LEM) containing reverse micelles could be successfully used for the downstream processing of lipase from Aspergillus niger. In the present work, we have attempted to increase the extraction and purification fold of lipase by using mixed reverse micelles (MRM) consisting of cat-ionic and nonionic surfactants in LEM. It was basically prepared by addition of the internal aqueous phase solution to the organic phase followed by the redispersion of the emulsion in the feed phase containing enzyme, which resulted in globules of water-oil-water (WOW) emulsion for the extraction of lipase. The optimum conditions for maximum lipase recovery (100%) and purification fold (17.0-fold) were CTAB concentration 0.075 M, Tween 80 concentration 0.012 M, at stirring speed of 500 rpm, contact time 15 min, internal aqueous phase pH 7, feed pH 9, KCl concentration 1 M, NaCl concentration 0.1 M, and ratio of membrane emulsion to feed volume 1:1. Incorporation of the nonionic surfactant (e.g., Tween 80) resulted in remarkable improvement in the purification fold (3.1-17.0) of the lipase. LEM containing a mixture of nonionic and cationic surfactants can be successfully used for the enhancement in the activity recovery and purification fold during downstream processing of enzymes/proteins.
机译：我们的早期工作首次证明了含有反胶束的液态乳状液膜（LEM）可成功用于黑曲霉脂肪酶的下游加工。在目前的工作中，我们尝试通过在LEM中使用由阳离子和非离子表面活性剂组成的混合反胶束（MRM）来增加脂肪酶的提取和纯化倍数。它的基本制备方法是将内部水相溶液添加到有机相中，然后将乳液重新分散在含有酶的进料相中，从而产生用于提取脂肪酶的水-油-水（WOW）乳液小球。最大脂肪酶回收率（100％）和纯化倍数（17.0倍）的最佳条件是CTAB浓度0.075 M，吐温80浓度0.012 M，搅拌速度为500 rpm，接触时间15分钟，内部水相pH 7，进料pH值为9，KCl浓度为1 M，NaCl浓度为0.1 M，膜乳液与进料量的比例为1：1。掺入非离子表面活性剂（例如吐温80）导致脂肪酶的提纯倍数（3.1-17.0）得到显着改善。包含非离子和阳离子表面活性剂混合物的LEM可成功用于增强酶/蛋白质下游加工过程中的活性回收率和纯化倍数。

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