Abstract In this work, enzymatically active polysaccharide-based hydrogels and aerogels have been developed. To this end, a thermostable β-galactosidase (TmLac) enzyme from Thermotoga maritima embedded in nanoflowers’ format was used to evaluate the capacity of the hydrogel matrices to preserve the hydrolytic activity of the enzyme and the reusability of the hydrogels formed. Commercial agar, unpurified agar and agarose were compared as supporting materials. Although the developed hydrogel capsules can be used at high temperature (75 °C) and reused for the digestion of lactose to a greater extent than the free nanoflowers, loaded hydrogel capsules behaved differently depending on the type of polysaccharide used. Commercial agar was the most promising one since these hydrogel capsules could be reused, maintaining the structural integrity and reaching higher enzymatic activity (after seven cycles at 75 °C) than the free TmLac-Ca2+ nanoflowers. To facilitate handling and storage, aerogels were developed by freeze-drying the hydrogel capsules. Aerogels of agarose and unpurified agar underwent structural changes during freeze-drying that adversely affected their subsequent use, losing their integrity after being rehydrated. However, commercial agar aerogels were successfully developed and reused thanks to the existing interactions with TmLac-Ca2+ nanoflowers (confirmed by FTIR), which resulted in better capsule integrity and enzyme protection. The hydrolytic activity of enzymatically active aerogels based on commercial agar was in the same range of the free TmLac and TmLac-Ca2+ nanoflowers, being significantly higher to their counterparts in the hydrated form (hydrogels based on commercial agar).

中文翻译：

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生物聚合物基质对酶活性水凝胶和载有 β-半乳糖苷酶纳米花的气凝胶水解乳糖的影响

摘要 在这项工作中，已开发出具有酶活性的多糖基水凝胶和气凝胶。为此，使用嵌入纳米花形式的海栖热袍菌中的热稳定 β-半乳糖苷酶 (TmLac) 来评估水凝胶基质的能力，以保持酶的水解活性和形成的水凝胶的可重复使用性。将商品琼脂、未纯化琼脂和琼脂糖作为支持材料进行比较。尽管开发的水凝胶胶囊可以在高温 (75 °C) 下使用，并且比游离的纳米花更大程度地用于消化乳糖，但加载的水凝胶胶囊根据所使用的多糖类型而表现不同。商业琼脂是最有前途的一种，因为这些水凝胶胶囊可以重复使用，保持结构完整性并达到比游离 TmLac-Ca2+ 纳米花更高的酶活性（在 75°C 下七个循环后）。为了便于处理和储存，通过冷冻干燥水凝胶胶囊来开发气凝胶。琼脂糖和未纯化琼脂的气凝胶在冷冻干燥过程中发生了结构变化，对其后续使用产生了不利影响，在再水化后失去了完整性。然而，由于与 TmLac-Ca2+ 纳米花的现有相互作用（由 FTIR 确认），商业琼脂气凝胶被成功开发和重复使用，从而导致更好的胶囊完整性和酶保护。基于商业琼脂的酶活性气凝胶的水解活性与游离 TmLac 和 TmLac-Ca2+ 纳米花的水解活性相同，