Expression and degradation patterns of chitinase purified from Xuehuali (Pyrus bretschneiderilia) pollen
| Autor: | Woo-Jin Jung, Jeong-An Cho, Chaeyeong Moon, Sang-Hyun Lee, Dong-Jun Seo, Yong-Su Song |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Chitin Biology 01 natural sciences Biochemistry Isozyme Gene Expression Regulation Enzymologic Substrate Specificity Pyrus 03 medical and health sciences chemistry.chemical_compound Hydrolysis Affinity chromatography Structural Biology Amino Acid Sequence Cloning Molecular Molecular Biology Peptide sequence Incubation chemistry.chemical_classification Chitinases General Medicine Oligosaccharide Isoenzymes carbohydrates (lipids) 030104 developmental biology chemistry Chitinase biology.protein Pollen 010606 plant biology & botany |
| Zdroj: | International Journal of Biological Macromolecules. 107:446-452 |
| ISSN: | 0141-8130 |
| DOI: | 10.1016/j.ijbiomac.2017.09.010 |
| Popis: | The present study investigated the expression pattern of chitinase in Xuehuali (Pyrus bretschneiderilia) pollen, as well as its subsequent degradation. The chitinase was purified and collected using chitin affinity column chromatography with regenerated chitin. After purification, four additional chitinase isozymes (chiA, chiB, chiC, and chiD) and chitinase (Chi II) were clearly expressed on SDS-PAGE gels that contained 0.01% glycol chitin. The chitinase reaction products were examined using GlcNAc, (GlcNAc)2, (GlcNAc)3, (GlcNAc)4, (GlcNAc)5, and (GlcNAc)6 as substrates at 2 and 24h after reaction via TLC and HPLC. The (GlcNAc)4 oligosaccharide was slightly degraded to (GlcNAc)2 after 24h of reaction with Xuehuali pollen chitinase on TLC. Meanwhile, (GlcNAc)5 was degraded to (GlcNAc)2-4, and 2300ppm (GlcNAc)6 was degraded to 246ppm (GlcNAc)2, 208ppm (GlcNAc)3, 572ppm (GlcNAc)4, and 336ppm (GlcNAc)5 on HPLC. With regard to temperature, the strongest Xuehuali pollen chitinase activity (0.69 unit/mL) was observed at 37°C after 3h of incubation, and with regard to pH, the strongest activity (0.72unit/mL) was observed at pH 3 after 3h of incubation. The main chitin oligomers degraded from (GlcNAc)6 were (GlcNAc)2 and (GlcNAc)4. |
| Databáze: | OpenAIRE |
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