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Int J Mol Sci. 2018 Jan 25;19(2). pii: E362. doi: 10.3390/ijms19020362.

Acidic Chitinase-Chitin Complex Is Dissociated in a Competitive Manner by Acetic Acid: Purification of Natural Enzyme for Supplementation Purposes.

Author information

1
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan. bm16024@ns.kogakuin.ac.jp.
2
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan. bu41265@ns.kogakuin.ac.jp.
3
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan. bd15003@ns.kogakuin.ac.jp.
4
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan. bt11532@ns.kogakuin.ac.jp.
5
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan. bt79310@ns.kogakuin.ac.jp.
6
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan. bt40522@ns.kogakuin.ac.jp.
7
RIKEN Center for Life Science Technologies, Tsurumi, Yokohama 230-0045, Japan. hideaki.shimizu@riken.jp.
8
Department of Clinical Biochemistry, Hematology and Immunology, Homolka Hospital, 142 20 Prague, Czech Republic. vaclav.matoska@homolka.cz.
9
Department of Clinical Biochemistry, Hematology and Immunology, Homolka Hospital, 142 20 Prague, Czech Republic. peter.bauer@bioinova.cz.
10
Bioinova Ltd., 142 20 Prague, Czech Republic. peter.bauer@bioinova.cz.
11
Department of Chemistry and Life Science, Kogakuin University, Hachioji, Tokyo 192-0015, Japan. f-oyama@cc.kogakuin.ac.jp.

Abstract

Acidic chitinase (Chia) has been implicated in asthma, allergic inflammations, and food processing. We have purified Chia enzymes with striking acid stability and protease resistance from chicken and pig stomach tissues using a chitin column and 8 M urea (urea-Chia). Here, we report that acetic acid is a suitable agent for native Chia purification from the stomach tissues using a chitin column (acetic acid-Chia). Chia protein can be eluted from a chitin column using 0.1 M acetic acid (pH 2.8), but not by using Gly-HCl (pH 2.5) or sodium acetate (pH 4.0 or 5.5). The melting temperatures of Chia are not affected substantially in the elution buffers, as assessed by differential scanning fluorimetry. Interestingly, acetic acid appears to be more effective for Chia-chitin dissociation than do other organic acids with similar structures. We propose a novel concept of this dissociation based on competitive interaction between chitin and acetic acid rather than on acid denaturation. Acetic acid-Chia also showed similar chitinolytic activity to urea-Chia, indicating that Chia is extremely stable against acid, proteases, and denaturing agents. Both acetic acid- and urea-Chia seem to have good potential for supplementation or compensatory purposes in agriculture or even biomedicine.

KEYWORDS:

GlcNAc; acetic acid; acidic chitinase; chitin; chitin column; competitive manner; natural enzyme; supplementation purposes; therapeutic agents; urea

PMID:
29370114
PMCID:
PMC5855584
DOI:
10.3390/ijms19020362
[Indexed for MEDLINE]
Free PMC Article

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