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Donor substrate promiscuity of the N-acetylglucosaminyltransferase activities of Pasteurella multocida heparosan synthase 2 (PmHS2) and Escherichia coli K5 KfiA.
Li, Yanhong; Yu, Hai; Thon, Vireak; Chen, Yi; Muthana, Musleh M; Qu, Jingyao; Hie, Liana; Chen, Xi.
Afiliação
  • Li Y; Department of Chemistry, University of California-Davis, One Shields Avenue, Davis, CA, 95616, USA.
Appl Microbiol Biotechnol ; 98(3): 1127-34, 2014 Feb.
Article em En | MEDLINE | ID: mdl-23661084
ABSTRACT
The biological activities of heparan sulfate (HS) and heparin (HP) are closely related to their molecular structures. Both Pasteurella multocida heparosan synthase 2 (PmHS2) and Escherichia coli K5 KfiA have been used for enzymatic and chemoenzymatic synthesis of HS and HP oligosaccharides and their derivatives. We show here that cloning using the pET15b vector and expressing PmHS2 as an N-His6-tagged fusion protein improve its expression level in E. coli. Investigation of the donor substrate specificity of the N-acetylglucosaminyltransferase activities of P. multocida heparosan synthase 2 (PmHS2) and E. coli K5 KfiA indicates the substrate promiscuities of PmHS2 and KfiA. Overall, both PmHS2 and KfiA can use uridine 5'-diphosphate-N-acetylglucosamine (UDP-GlcNAc) and some of its C2'- and C6'-derivatives as donor substrates for their α1-4-GlcNAcT activities. Nevertheless, PmHS2 has a broader tolerance towards substrate modifications. Other than the UDP-sugars that can be used by KfiA, additional C6'-derivatives of UDP-GlcNAc, UDP-glucose, and UDP-N-acetylgalactosamine (UDP-GalNAc) are tolerable substrates for the α1-4-GlcNAcT activity of PmHS2. The substrate promiscuities of PmHS2 and KfiA will allow efficient chemoenzymatic synthesis of diverse HS and HP oligosaccharide derivatives which may have improved or altered activities compared to their natural counterparts.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pasteurella multocida / Glicosiltransferases / N-Acetilglucosaminiltransferases / Proteínas de Escherichia coli / Escherichia coli Idioma: En Revista: Appl Microbiol Biotechnol Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pasteurella multocida / Glicosiltransferases / N-Acetilglucosaminiltransferases / Proteínas de Escherichia coli / Escherichia coli Idioma: En Revista: Appl Microbiol Biotechnol Ano de publicação: 2014 Tipo de documento: Article País de afiliação: Estados Unidos