Abstract

Chitooligosaccharides (COS) have a great potential to be used by pharmaceutical industry due to their many biological activities. The use of enzymes to produce them is very advantageous, however it still faces many challenges, such as discovering new strains capable to produce enzymes that are able to generate bioactive oligosaccharides. In the present study a purification protein protocol was performed to purify chitosanases produced by Bacillus toyonensis CCT 7899 for further chitosan hydrolysis. The produced chitooligosaccharides were characterized by mass spectroscopy (MS) and their antiedematogenic effect was investigated through carrageenan-induced paw edema model. The animals were treated previously to inflammation by intragastric route with COS at 30, 300 and 600 mg/kg. The purification protocol showed a good performance for the chitosanases purification using 0.20 M NaCl solution to elute it, with a 9.54-fold purification factor. The treatment with COS promoted a decrease of paw edema at all evaluated times and the AUC0–4h, proving that COS produced showed activity in acute inflammation like commercial anti-inflammatory Dexamethasone (corticosteroid). Therefore, the strategy used to purification was successfully applied and it was possible to generate bioactive oligosaccharides with potential pharmacological use.

摘要

壳寡糖 (COS) 因其多种生物活性而具有巨大的医药工业应用潜力。使用酶来生产它们是非常有利的，但是它仍然面临许多挑战，例如发现能够产生能够产生生物活性寡糖的酶的新菌株。在本研究中，进行了纯化蛋白方案以纯化丰盈芽孢杆菌产生的壳聚糖酶CCT 7899 用于进一步的壳聚糖水解。所产生的壳寡糖通过质谱（MS）进行表征，并通过角叉菜胶诱导的爪水肿模型研究其抗水肿作用。先前通过胃内途径用 30、300 和 600 mg/kg 的 COS 对动物进行炎症治疗。纯化方案显示使用 0.20 M NaCl 溶液洗脱壳聚糖酶具有良好的性能，纯化因子为 9.54 倍。COS 治疗促进了所有评估时间和 AUC0-4h 的爪子水肿减少，证明 COS 产生的急性炎症表现出活性，如商业抗炎地塞米松（皮质类固醇）。所以，