Acidic Chitinase-Chitin Complex Is Dissociated in a Competitive Manner by Acetic Acid: Purification of Natural Enzyme for Supplementation Purposes
Autor: | Hideaki Shimizu, Eri Tabata, Fumitaka Oyama, Vaclav Matoska, Yasutada Imamura, Masayoshi Sakaguchi, Yasusato Sugahara, Satoshi Wakita, Peter O. Bauer, Akinori Kashimura |
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Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Swine GlcNAc acetic acid acidic chitinase chitin chitin column competitive manner natural enzyme supplementation purposes therapeutic agents urea medicine.medical_treatment Chitin Catalysis Dissociation (chemistry) Article lcsh:Chemistry Inorganic Chemistry 03 medical and health sciences chemistry.chemical_compound Acetic acid medicine Animals Physical and Theoretical Chemistry lcsh:QH301-705.5 Molecular Biology Spectroscopy chemistry.chemical_classification Protease Chromatography biology Chemistry Organic Chemistry Chitinases Stomach General Medicine Computer Science Applications 030104 developmental biology Enzyme lcsh:Biology (General) lcsh:QD1-999 Chitinase Urea biology.protein Sodium acetate Chickens Protein Binding |
Zdroj: | International Journal of Molecular Sciences; Volume 19; Issue 2; Pages: 362 International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 19, Iss 2, p 362 (2018) |
ISSN: | 1422-0067 |
DOI: | 10.3390/ijms19020362 |
Popis: | 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. |
Databáze: | OpenAIRE |
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