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Substrate Promiscuity of N-Acetylhexosamine 1-Kinases
N-Acetylhexosamine 1-kinase (NahK) catalyzes the direct addition of a phosphate from adenosine 5'-triphosphate (ATP) to the anomeric position of N-acetylhexosamine and shows similar activity towards N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc). Herein we report the cloning, c...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6264712/ https://www.ncbi.nlm.nih.gov/pubmed/21799473 http://dx.doi.org/10.3390/molecules16086396 |
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author | Li, Yanhong Yu, Hai Chen, Yi Lau, Kam Cai, Li Cao, Hongzhi Tiwari, Vinod Kumar Qu, Jingyao Thon, Vireak Wang, Peng George Chen, Xi |
author_facet | Li, Yanhong Yu, Hai Chen, Yi Lau, Kam Cai, Li Cao, Hongzhi Tiwari, Vinod Kumar Qu, Jingyao Thon, Vireak Wang, Peng George Chen, Xi |
author_sort | Li, Yanhong |
collection | PubMed |
description | N-Acetylhexosamine 1-kinase (NahK) catalyzes the direct addition of a phosphate from adenosine 5'-triphosphate (ATP) to the anomeric position of N-acetylhexosamine and shows similar activity towards N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc). Herein we report the cloning, characterization, and substrate specificity studies of two NahKs from Bifidobacterium infantis ATCC15697 and Bifidobacterium longum ATCC55813, respectively. A new capillary electrophoresis assay method has been developed for enzyme activity assays. Both enzymes have a good expression level in E. coli (180–185 mg/L culture) and can tolerate diverse modifications at C2 of GlcNAc and GalNAc. Various GlcNAc derivatives with C6, both C2 and C6, as well as both C2 and C3 modifications are tolerable substrates for the newly cloned NahKs. Quite interestingly, despite of their low activities toward glucose and galactose, the activities of both NahKs are much higher for mannose and some of its C2, C4, and C6 derivatives. These NahKs are excellent catalysts for enzymatic and chemoenzymatic synthesis of carbohydrates. |
format | Online Article Text |
id | pubmed-6264712 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-62647122018-12-10 Substrate Promiscuity of N-Acetylhexosamine 1-Kinases Li, Yanhong Yu, Hai Chen, Yi Lau, Kam Cai, Li Cao, Hongzhi Tiwari, Vinod Kumar Qu, Jingyao Thon, Vireak Wang, Peng George Chen, Xi Molecules Article N-Acetylhexosamine 1-kinase (NahK) catalyzes the direct addition of a phosphate from adenosine 5'-triphosphate (ATP) to the anomeric position of N-acetylhexosamine and shows similar activity towards N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc). Herein we report the cloning, characterization, and substrate specificity studies of two NahKs from Bifidobacterium infantis ATCC15697 and Bifidobacterium longum ATCC55813, respectively. A new capillary electrophoresis assay method has been developed for enzyme activity assays. Both enzymes have a good expression level in E. coli (180–185 mg/L culture) and can tolerate diverse modifications at C2 of GlcNAc and GalNAc. Various GlcNAc derivatives with C6, both C2 and C6, as well as both C2 and C3 modifications are tolerable substrates for the newly cloned NahKs. Quite interestingly, despite of their low activities toward glucose and galactose, the activities of both NahKs are much higher for mannose and some of its C2, C4, and C6 derivatives. These NahKs are excellent catalysts for enzymatic and chemoenzymatic synthesis of carbohydrates. MDPI 2011-07-28 /pmc/articles/PMC6264712/ /pubmed/21799473 http://dx.doi.org/10.3390/molecules16086396 Text en © 2011 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 license (http://creativecommons.org/licenses/by/3.0/). |
spellingShingle | Article Li, Yanhong Yu, Hai Chen, Yi Lau, Kam Cai, Li Cao, Hongzhi Tiwari, Vinod Kumar Qu, Jingyao Thon, Vireak Wang, Peng George Chen, Xi Substrate Promiscuity of N-Acetylhexosamine 1-Kinases |
title | Substrate Promiscuity of N-Acetylhexosamine 1-Kinases |
title_full | Substrate Promiscuity of N-Acetylhexosamine 1-Kinases |
title_fullStr | Substrate Promiscuity of N-Acetylhexosamine 1-Kinases |
title_full_unstemmed | Substrate Promiscuity of N-Acetylhexosamine 1-Kinases |
title_short | Substrate Promiscuity of N-Acetylhexosamine 1-Kinases |
title_sort | substrate promiscuity of n-acetylhexosamine 1-kinases |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6264712/ https://www.ncbi.nlm.nih.gov/pubmed/21799473 http://dx.doi.org/10.3390/molecules16086396 |
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