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从蓝蟹内脏中制备粗壳聚糖酶及其在虾壳壳聚糖生物活性寡糖生产中的应用。

Preparation of a crude chitosanase from blue crab viscera as well as its application in the production of biologically active chito-oligosaccharides from shrimp shells chitosan.

机构信息

Laboratory of Enzyme Engineering and Microbiology, National School of Engineering of Sfax (ENIS), University of Sfax, P.O. Box 1173, Sfax 3038, Tunisia.

Department of Chemistry in Pharmaceutical Science, Faculty of Pharmacy, Pluridisciplinar Institute, Complutense University of Madrid, 28040 Madrid, Spain.

出版信息

Int J Biol Macromol. 2019 Oct 15;139:558-569. doi: 10.1016/j.ijbiomac.2019.07.116. Epub 2019 Jul 21.

Abstract

In this study, a digestive chitosanase from blue crab (Portunus segnis) viscera was extracted, characterized and applied. The crude chitosanase showed optimum activity at pH 4.0 and 60 °C and retained >80% of its activity over a pH range from 3.0 to 10.0. Subsequently, the crude chitosanase was applied to produce bioactive varying molecular weight (M) and acetylation degree chitosan-depolymerization products (CDP) with specially sequences composition determined by MALDI-TOF MS owing to an endo-cleavage mode. This hydrolysis process allowed to the preparation, after 24 h of incubation at 40 °C, of a low Mw water soluble CDP (H <4.4 kDa) with DP up to 6 and a high Mw CDP (C 142.19 kDa). Following their physicochemical characterization, the functional properties, antioxidant and antimicrobial activities of CDP were investigated. Interestingly, as compared to the native chitosan, CDP, especially low M derivatives (H ) exhibited potent antioxidant activities, while high M derivatives, especially C , markedly inhibited the growth of all tested bacteria and fungi. These results may provide novel insights into the efficiency of chitosan depolymerisation using the Portunus segnis digestive crude chitosanase as a simple, inexpensive and easily method to produce bioactive chitosan-derivatives and that this bioactivity depends highly on their attractive characteristics.

摘要

在这项研究中,从蓝蟹(Portunus segnis)内脏中提取、表征和应用了一种消化甲壳质酶。粗甲壳质酶在 pH 4.0 和 60°C 下显示出最佳活性,并在 pH 3.0 至 10.0 范围内保持>80%的活性。随后,将粗甲壳质酶应用于生产具有特殊序列组成的生物活性、不同分子量(Mw)和乙酰化度的壳聚糖解聚产物(CDP),这归因于内切裂解模式。由于这种水解过程,在 40°C 孵育 24 小时后,可以制备Mw 低的水溶性 CDP(H <4.4 kDa),其 DP 高达 6,以及 Mw 高的 CDP(C 142.19 kDa)。在对其理化特性进行表征后,研究了 CDP 的功能特性、抗氧化和抗菌活性。有趣的是,与天然壳聚糖相比,CDP,尤其是低 M 衍生物(H )表现出很强的抗氧化活性,而高 M 衍生物,特别是 C ,显著抑制了所有测试细菌和真菌的生长。这些结果可能为利用蓝蟹消化粗甲壳质酶作为一种简单、廉价和容易的方法生产生物活性壳聚糖衍生物提供新的见解,并且这种生物活性高度依赖于它们吸引人的特性。

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