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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...

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Autores principales: Li, Yanhong, Yu, Hai, Chen, Yi, Lau, Kam, Cai, Li, Cao, Hongzhi, Tiwari, Vinod Kumar, Qu, Jingyao, Thon, Vireak, Wang, Peng George, Chen, Xi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2011
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.
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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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