Compared to standard protein formulations like aqueous solutions, crystalline proteins may offer superior properties (e.g., higher purity and concentration, reduced storage costs, and enhanced shelf life). In this work, crystallization conditions for lysozyme from Gallus gallus and a lipase from Thermomyces lanuginosus were characterized in microbatch experiments. The previously described positive effects of water-soluble substituted alkylammonium-based ionic liquids as additives on the crystallization of these enzymes (e.g., faster crystal growth kinetics and the formation of larger, sturdier crystals) was confirmed. With the use of optimized conditions, the crystallization processes were transferred into parallel-operated stirred crystallizers on a 5 mL scale. A higher yield and faster crystal growth kinetics were observed when using additives. For lysozyme, a yield of 97% was obtained within 2 h. For lipase, a yield of 95% was obtained within 2 h by stepwise addition of 50 g L~(-1) PEG 10000. The crystallization processes were successfully scaled-up into geometrically similar stirred crystallizers on a 100 mL and 1 L scale, respectively. Favorable crystal morphologies and adequate crystal size distributions were obtained. Unfavorable substances were removed from the crystals by washing.
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机译：与诸如水溶液的标准蛋白质制剂相比，结晶蛋白质可以提供优越的性能（例如，更高的纯度和浓度，降低的存储成本和更长的保存期限）。在这项工作中，在微分批实验中表征了来自鸡鸡的溶菌酶和来自嗜热单胞菌的脂肪酶的结晶条件。确认了前述作为添加剂的水溶性取代的烷基铵基离子液体对这些酶的结晶的积极作用（例如，更快的晶体生长动力学和更大，更结实的晶体的形成）。通过使用优化的条件，将结晶过程转移到5 mL规模的平行操作的搅拌结晶器中。当使用添加剂时，观察到更高的产率和更快的晶体生长动力学。对于溶菌酶，在2小时内获得97％的产率。对于脂肪酶，通过逐步添加50 g L〜（-1）PEG 10000，在2小时内可获得95％的收率。成功地将结晶过程按比例放大至100 mL和1 L规模的几何相似的搅拌结晶器，分别。获得了有利的晶体形态和足够的晶体尺寸分布。通过洗涤从晶体中除去不利的物质。
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