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木瓜蛋白酶固定在异功能膜细菌纤维素上作为一种皮肤伤口清创的潜在策略。

Papain immobilization on heterofunctional membrane bacterial cellulose as a potential strategy for the debridement of skin wounds.

机构信息

Universidade Federal do Ceará (UFC), Departamento de Química Orgânica e Inorgânica, Bloco 935, 60455-760 Fortaleza, Ceará, Brazil.

Universidade Federal do Ceará (UFC), Central Analítica, Bloco 928, 60455-970 Fortaleza, Ceará, Brazil.

出版信息

Int J Biol Macromol. 2020 Dec 15;165(Pt B):3065-3077. doi: 10.1016/j.ijbiomac.2020.10.200. Epub 2020 Oct 28.

Abstract

We combined the chemical and physical methods of papain immobilization through the aldehyde groups available on oxidized bacterial cellulose (OxBC) to provide high proteolytic activity for future applications as bioactive dressing. Bacterial cellulose (BC) was obtained by the fermentation of Komagataeibacter hansenii in Hestrin-Schramm medium for 5 days, followed by purification and oxidation using NaIO. Surface response methodology was used to optimize papain immobilization (2%, w/v) for 24 h. The independent variables: pH (3-7) and temperature (5 to 45 °C) were investigated. The mathematically validated optimal conditions of 45 °C and pH 7 had a statistical effect on the immobilization yield (IY) of papain in OxBC (52.9%). These ideal conditions were also used for papain immobilization in BC (unoxidized). The IY of 9.1% was lower than that of OxBC. OxBC-Papain and BC-Papain were investigated using thermal analysis, confocal microscopy, and diffusion testing. The OxBC support exhibited a more interactive chemical structure than the BC support, and was capable of immobilizing papain by covalent bonds (-C-NHR) and adsorption (ion exchange), with 93.3% recovered activity, 49.4% immobilization efficiency, and better thermal stability. Papain immobilized to OxBC by adsorption displayed 53% widespread papain activity. The results indicate the potential of prolonged bioactivity in debrided chronic wounds.

摘要

我们通过氧化细菌纤维素 (OxBC) 上的醛基结合了木瓜蛋白酶的化学和物理固定化方法,为未来用作生物活性敷料提供了高蛋白酶活性。细菌纤维素 (BC) 通过在 Hestrin-Schramm 培养基中发酵 Komagataeibacter hansenii 获得,持续 5 天,然后使用 NaIO 进行纯化和氧化。表面响应方法用于优化木瓜蛋白酶固定化(2%,w/v)24 小时。研究了独立变量:pH(3-7)和温度(5 至 45°C)。经过数学验证的最佳条件为 45°C 和 pH 7,对 OxBC 中木瓜蛋白酶的固定化产率(IY)具有统计学影响(52.9%)。这些理想条件也用于未氧化的 BC 中的木瓜蛋白酶固定化。IY 为 9.1%,低于 OxBC。使用热分析、共聚焦显微镜和扩散测试研究了 OxBC-Papain 和 BC-Papain。OxBC 载体表现出比 BC 载体更具交互性的化学结构,能够通过共价键(-C-NHR)和吸附(离子交换)固定化木瓜蛋白酶,具有 93.3%的回收活性、49.4%的固定化效率和更好的热稳定性。通过吸附固定到 OxBC 的木瓜蛋白酶显示出 53%的广泛木瓜蛋白酶活性。结果表明在清创慢性伤口中具有延长生物活性的潜力。

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