NahK/GlmU fusion enzyme: characterization and one-step enzymatic synthesis of UDP-N-acetylglucosamine.
Biotechnol Lett
; 34(7): 1321-6, 2012 Jul.
Article
em En
| MEDLINE
| ID: mdl-22456903
ABSTRACT
The availability of uridine 5'-diphosphate N-acetylglucosamine (UDP-GlcNAc) is a prerequisite for the GlcNAc-transferase-catalyzed glycosylation reaction. UDP-GlcNAc has already been synthesized using an N-acetylhexosamine 1-kinase (NahK) and a GlcNAc-1-P uridyltransferase (truncated GlmU) and here, a fusion enzyme was constructed with truncated GlmU and NahK. After determination of the optimum catalytic condition (pH 8.0 at 40 °C), the fusion enzyme was used to synthesize UDP-GlcNAc in a single step with a yield of 88 % from GlcNAc, ATP and UTP. Furthermore, a simplified purification method was demonstrated using separation by gel filtration after by-product digestion with alkaline phosphatase. An overall yield of 77 % and a purity of over 90 % were achieved.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Uridina Difosfato N-Acetilglicosamina
/
Biotecnologia
/
Enzimas
Idioma:
En
Revista:
Biotechnol Lett
Ano de publicação:
2012
Tipo de documento:
Article