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Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14
Glycosaminoglycan (GAG) lyase is an effective tool for the structural and functional studies of glycosaminoglycans and preparation of functional oligosaccharides. A new GAG lyase from Microbacterium sp. H14 was cloned, expressed, purified, and characterized, with a molecular weight of approximately...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6950261/ https://www.ncbi.nlm.nih.gov/pubmed/31810166 http://dx.doi.org/10.3390/md17120681 |
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author | Sun, Junhao Han, Xu Song, Guanrui Gong, Qianhong Yu, Wengong |
author_facet | Sun, Junhao Han, Xu Song, Guanrui Gong, Qianhong Yu, Wengong |
author_sort | Sun, Junhao |
collection | PubMed |
description | Glycosaminoglycan (GAG) lyase is an effective tool for the structural and functional studies of glycosaminoglycans and preparation of functional oligosaccharides. A new GAG lyase from Microbacterium sp. H14 was cloned, expressed, purified, and characterized, with a molecular weight of approximately 85.9 kDa. The deduced lyase HCLaseM belonged to the polysaccharide lyase (PL) family 8. Based on the phylogenetic tree, HCLaseM could not be classified into the existing three subfamilies of this family. HCLaseM showed almost the same enzyme activity towards hyaluronan (HA), chondroitin sulfate A (CS-A), CS-B, CS-C, and CS-D, which was different from reported GAG lyases. HCLaseM exhibited the highest activities to both HA and CS-A at its optimal temperature (35 °C) and pH (pH 7.0). HCLaseM was stable in the range of pH 5.0–8.0 and temperature below 30 °C. The enzyme activity was independent of divalent metal ions and was not obviously affected by most metal ions. HCLaseM is an endo-type enzyme yielding unsaturated disaccharides as the end products. The facilitated diffusion effect of HCLaseM is dose-dependent in animal experiments. These properties make it a candidate for further basic research and application. |
format | Online Article Text |
id | pubmed-6950261 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-69502612020-01-16 Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14 Sun, Junhao Han, Xu Song, Guanrui Gong, Qianhong Yu, Wengong Mar Drugs Article Glycosaminoglycan (GAG) lyase is an effective tool for the structural and functional studies of glycosaminoglycans and preparation of functional oligosaccharides. A new GAG lyase from Microbacterium sp. H14 was cloned, expressed, purified, and characterized, with a molecular weight of approximately 85.9 kDa. The deduced lyase HCLaseM belonged to the polysaccharide lyase (PL) family 8. Based on the phylogenetic tree, HCLaseM could not be classified into the existing three subfamilies of this family. HCLaseM showed almost the same enzyme activity towards hyaluronan (HA), chondroitin sulfate A (CS-A), CS-B, CS-C, and CS-D, which was different from reported GAG lyases. HCLaseM exhibited the highest activities to both HA and CS-A at its optimal temperature (35 °C) and pH (pH 7.0). HCLaseM was stable in the range of pH 5.0–8.0 and temperature below 30 °C. The enzyme activity was independent of divalent metal ions and was not obviously affected by most metal ions. HCLaseM is an endo-type enzyme yielding unsaturated disaccharides as the end products. The facilitated diffusion effect of HCLaseM is dose-dependent in animal experiments. These properties make it a candidate for further basic research and application. MDPI 2019-12-02 /pmc/articles/PMC6950261/ /pubmed/31810166 http://dx.doi.org/10.3390/md17120681 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Sun, Junhao Han, Xu Song, Guanrui Gong, Qianhong Yu, Wengong Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14 |
title | Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14 |
title_full | Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14 |
title_fullStr | Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14 |
title_full_unstemmed | Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14 |
title_short | Cloning, Expression, and Characterization of a New Glycosaminoglycan Lyase from Microbacterium sp. H14 |
title_sort | cloning, expression, and characterization of a new glycosaminoglycan lyase from microbacterium sp. h14 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6950261/ https://www.ncbi.nlm.nih.gov/pubmed/31810166 http://dx.doi.org/10.3390/md17120681 |
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