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Mar. Drugs 2019, 17(1), 68; https://doi.org/10.3390/md17010068

Design and Synthesis of a Chitodisaccharide-Based Affinity Resin for Chitosanases Purification

1
Department of Pharmacology, College of basic Medicine, Qingdao University, Qingdao 266071, China
2
Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
*
Authors to whom correspondence should be addressed.
Received: 22 December 2018 / Revised: 14 January 2019 / Accepted: 15 January 2019 / Published: 21 January 2019
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Abstract

Chitooligosaccharides (CHOS) have gained increasing attention because of their important biological activities. Enhancing the efficiency of CHOS production essentially requires screening of novel chitosanase with unique characteristics. Therefore, a rapid and efficient one-step affinity purification procedure plays important roles in screening native chitosanases. In this study, we report the design and synthesis of affinity resin for efficient purification of native chitosanases without any tags, using chitodisaccharides (CHDS) as an affinity ligand, to couple with Sepharose 6B via a spacer, cyanuric chloride. Based on the CHDS-modified affinity resin, a one-step affinity purification method was developed and optimized, and then applied to purify three typical glycoside hydrolase (GH) families: 46, 75, and 80 chitosanase. The three purified chitosanases were homogeneous with purities of greater than 95% and bioactivity recovery of more than 40%. Moreover, we also developed a rapid and efficient affinity purification procedure, in which tag-free chitosanase could be directly purified from supernatant of bacterial culture. The purified chitosanases samples using such a procedure had apparent homogeneity, with more than 90% purity and 10–50% yield. The novel purification methods established in this work can be applied to purify native chitosanases in various scales, such as laboratory and industrial scales. View Full-Text
Keywords: chitosanases; adsorption analysis; affinity purification chitosanases; adsorption analysis; affinity purification
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Li, S.; Wang, L.; Chen, X.; Sun, M.; Han, Y. Design and Synthesis of a Chitodisaccharide-Based Affinity Resin for Chitosanases Purification. Mar. Drugs 2019, 17, 68.

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