Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan Oligosaccharides
Partially acetylated chitosan oligosaccharides (COS), which consists of N-acetylglucosamine (GlcNAc) and glucosamine (GlcN) residues, is a structurally complex biopolymer with a variety of biological activities. Therefore, it is challenging to elucidate acetylation patterns and the molecular structu...
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| Format: | Article |
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MDPI AG
2019-01-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/17/2/74 |
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| author | Xian-Yu Zhu Yong Zhao Huai-Dong Zhang Wen-Xia Wang Hai-Hua Cong Heng Yin |
| author_facet | Xian-Yu Zhu Yong Zhao Huai-Dong Zhang Wen-Xia Wang Hai-Hua Cong Heng Yin |
| author_sort | Xian-Yu Zhu |
| collection | DOAJ |
| description | Partially acetylated chitosan oligosaccharides (COS), which consists of N-acetylglucosamine (GlcNAc) and glucosamine (GlcN) residues, is a structurally complex biopolymer with a variety of biological activities. Therefore, it is challenging to elucidate acetylation patterns and the molecular structure-function relationship of COS. Herein, the detailed deacetylation pattern of chitin deacetylase from <i>Saccharomyces cerevisiae</i>, <i>Sc</i>CDA<sub>2</sub>, was studied. Which solves the randomization of acetylation patterns during COS produced by chemical. <i>Sc</i>CDA<sub>2</sub> also exhibits about 8% and 20% deacetylation activity on crystalline chitin and colloid chitin, respectively. Besides, a method for separating and detecting partially acetylated chitosan oligosaccharides by high performance liquid chromatography and electrospray ionization mass spectrometry (HPLC-ESI-MS) system has been developed, which is fast and convenient, and can be monitored online. Mass spectrometry sequencing revealed that <i>Sc</i>CDA<sub>2</sub> produced COS with specific acetylation patterns of DAAA, ADAA, AADA, DDAA, DADA, ADDA and DDDA, respectively. <i>Sc</i>CDA<sub>2</sub> does not deacetylate the GlcNAc unit that is closest to the reducing end of the oligomer furthermore <i>Sc</i>CDA<sub>2</sub> has a multiple-attack deacetylation mechanism on chitin oligosaccharides. This specific mode of action significantly enriches the existing limited library of chitin deacetylase deacetylation patterns. This fully defined COS may be used in the study of COS structure and function. |
| first_indexed | 2024-04-13T00:46:21Z |
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| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-04-13T00:46:21Z |
| publishDate | 2019-01-01 |
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| series | Marine Drugs |
| spelling | doaj.art-23842d1dc82f4d81a3bd4bcb32deeed82022-12-22T03:10:00ZengMDPI AGMarine Drugs1660-33972019-01-011727410.3390/md17020074md17020074Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan OligosaccharidesXian-Yu Zhu0Yong Zhao1Huai-Dong Zhang2Wen-Xia Wang3Hai-Hua Cong4Heng Yin5Liaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, ChinaLiaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, ChinaLiaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, ChinaLiaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, ChinaCollege of Food Science and Engineering, Dalian Ocean University, Dalian 116023, ChinaLiaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, ChinaPartially acetylated chitosan oligosaccharides (COS), which consists of N-acetylglucosamine (GlcNAc) and glucosamine (GlcN) residues, is a structurally complex biopolymer with a variety of biological activities. Therefore, it is challenging to elucidate acetylation patterns and the molecular structure-function relationship of COS. Herein, the detailed deacetylation pattern of chitin deacetylase from <i>Saccharomyces cerevisiae</i>, <i>Sc</i>CDA<sub>2</sub>, was studied. Which solves the randomization of acetylation patterns during COS produced by chemical. <i>Sc</i>CDA<sub>2</sub> also exhibits about 8% and 20% deacetylation activity on crystalline chitin and colloid chitin, respectively. Besides, a method for separating and detecting partially acetylated chitosan oligosaccharides by high performance liquid chromatography and electrospray ionization mass spectrometry (HPLC-ESI-MS) system has been developed, which is fast and convenient, and can be monitored online. Mass spectrometry sequencing revealed that <i>Sc</i>CDA<sub>2</sub> produced COS with specific acetylation patterns of DAAA, ADAA, AADA, DDAA, DADA, ADDA and DDDA, respectively. <i>Sc</i>CDA<sub>2</sub> does not deacetylate the GlcNAc unit that is closest to the reducing end of the oligomer furthermore <i>Sc</i>CDA<sub>2</sub> has a multiple-attack deacetylation mechanism on chitin oligosaccharides. This specific mode of action significantly enriches the existing limited library of chitin deacetylase deacetylation patterns. This fully defined COS may be used in the study of COS structure and function.https://www.mdpi.com/1660-3397/17/2/74chitin deacetylasedeacetylation patternschitooligosaccharidesseparatingdetecting |
| spellingShingle | Xian-Yu Zhu Yong Zhao Huai-Dong Zhang Wen-Xia Wang Hai-Hua Cong Heng Yin Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan Oligosaccharides Marine Drugs chitin deacetylase deacetylation patterns chitooligosaccharides separating detecting |
| title | Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan Oligosaccharides |
| title_full | Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan Oligosaccharides |
| title_fullStr | Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan Oligosaccharides |
| title_full_unstemmed | Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan Oligosaccharides |
| title_short | Characterization of the Specific Mode of Action of a Chitin Deacetylase and Separation of the Partially Acetylated Chitosan Oligosaccharides |
| title_sort | characterization of the specific mode of action of a chitin deacetylase and separation of the partially acetylated chitosan oligosaccharides |
| topic | chitin deacetylase deacetylation patterns chitooligosaccharides separating detecting |
| url | https://www.mdpi.com/1660-3397/17/2/74 |
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