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首页> 外文期刊>RSC Advances >Solid liquid liquid extraction of porcine gastric mucins from homogenized animal material
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Solid liquid liquid extraction of porcine gastric mucins from homogenized animal material

机译：从均质动物材料中固液提取猪胃粘蛋白

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Mucins purified from porcine stomachs have gained importance in biomedical applications as they exhibit unique features such as hydrogel formation, lubricity and antivirality. Because commercially available porcine gastric mucins (PGM) are neither gel forming, nor do they reduce friction, a robust purification process for functional mucins from porcine tissue (mainly Muc5AC) is necessary. Based on our former investigations (V. J. Sch?mig, B. T. K?sdorf, C. Scholz, K. Bidmon, O. Lieleg and S. Berensmeier, RSC Adv., 2016, 6, 44932–44943), we further optimized the established purification process in terms of productivity and overall yield as well as ease of scalability. We therefore introduced a novel extraction method – solid liquid liquid extraction (SLLE) – as an early capture step from homogenized porcine stomach, which combines conventional solid liquid extraction with a second, immiscible solvent, to simultaneously delipidate the tissue and extract hydrophilic proteins into the polar phase. Using Design of Experiments (DoE) the parameters incubation time and ratio of solvent phases (hexane/water) were identified to 3 h and 1/15 for SLLE, respectively. PGM was collected in the polar phase and further purified by size exclusion chromatography and diafiltration. With the homogenization of porcine stomach and the introduction of the SLLE, up to 3570 mg mucin per stomach can be purified, resulting in 55 times more PGM compared to the former published reference process. The productivity of the process was up to 25 mg mucin per stomach and per h, representing an improved process productivity by a factor of 4. Lubricity and gel formation of the purified mucin were retained with the optimized extraction protocol.
机译：从猪胃中纯化的粘蛋白具有独特的功能，例如水凝胶形成，润滑性和抗病毒性，因此在生物医学应用中变得越来越重要。由于市售的猪胃粘蛋白（PGM）既不会形成凝胶，也不会减少摩擦，因此必须有一个强大的从猪组织（主要是Muc5AC）中提取功能性粘蛋白的方法。基于我们之前的调查（VJ Sch？mig，BT K？sdorf，C。Scholz，K。Bidmon，O。Lieleg和S. Berensmeier， RSC Adv。，2016，6，44932–44943 ），我们在生产率，总产量以及可扩展性方面进一步优化了既定的纯化工艺。因此，我们引入了一种新颖的提取方法-固液提取（SLLE）-作为从均质猪胃中捕获的早期步骤，该方法将常规固液提取与不混溶的第二种溶剂相结合，同时使组织脱脂并从亲水蛋白中提取亲水蛋白。极性相。使用实验设计（DoE），SLLE的参数孵育时间和溶剂相（己烷/水）比例分别确定为3 h和1/15。在极性相中收集PGM，并通过尺寸排阻色谱法和渗滤进一步纯化。随着猪胃的均质化和SLLE的引入，每个胃中最多可纯化3570 mg粘蛋白，与以前公布的参考方法相比，其PGM增长55倍。该方法的生产率为每胃每小时每胃高达25 mg粘蛋白，这表明工艺生产率提高了4倍。纯化的粘蛋白的润滑性和凝胶形成通过优化的提取方案得以保留。

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