Skip Navigation


Journal of Biochemistry Advance Access originally published online on November 4, 2007
Journal of Biochemistry 2008 143(2):187-197; doi:10.1093/jb/mvm208
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
143/2/187    most recent
mvm208v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Tsukamoto, H.
Right arrow Articles by Yamamoto, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tsukamoto, H.
Right arrow Articles by Yamamoto, T.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2007 The Japanese Biochemical Society.

Photobacterium sp. JT-ISH-224 Produces Two Sialyltransferases, {alpha}-/β-Galactoside {alpha}2,3-Sialyltransferase and β-Galactoside {alpha}2,6-Sialyltransferase

Hiroshi Tsukamoto{dagger}, Yoshimitsu Takakura, Toshiki Mine and Takeshi Yamamoto*

Glycotechnology Business Unit, Japan Tobacco Inc., Higashibara, Iwata, Shizuoka 438-0802, Japan

*To whom correspondence should be addressed. Tel: +81-538-32-7389, Fax: +81-538-33-6046, E-mail: takeshi.yamamoto{at}ims.jti.co.jp.

{dagger}Correspondence may also be addressed: Tel: +81-538-32-7337, Fax: +81-538-33-6046, E-mail: hiroshi.tsukamoto{at}ims.jti.co.jp

Received August 21, 2007; Accepted October 22, 2007


  Abstract

A novel bacterium, Photobacterium sp. JT-ISH-224, that produces {alpha}-/β-galactoside {alpha}2,3-sialyltransferase and β-galactoside {alpha}2,6-sialyltransferase, was isolated from the gut of a Japanese barracuda. The genes that encode the enzymes were cloned from the genomic library of the bacterium using the genes encoding {alpha}-/β-galactoside {alpha}2,3-sialyltransferase from P. phosphoreum and β-galactoside {alpha}2,6-sialyltransferase from P. damselae as probes. The nucleotide sequences were determined, and open reading frames of 1,230 and 1,545 bp for encoding an {alpha}2,3-sialyltransferase and an {alpha}2,6-sialyltransferase of 409- and 514-amino acid residues, respectively, were identified. The {alpha}2,3-sialyltransferase had 92% amino acid sequence identity with the P. phosphoreum {alpha}2,3-sialyltransferase, whereas the {alpha}2,6-sialyltransferase had 54% amino acid sequence identity with the P. damselae {alpha}2,6-sialyltransferase. For both enzymes, the DNA fragments that encoded the full-length protein and its truncated form lacking the putative signal peptide sequence were amplified by a polymerase chain reaction and cloned into an expression vector. Each gene was expressed in Escherichia coli, and the lysate from each strain had enzymatic activity. The {alpha}2,3-sialyltransferase catalysed the transfer of N-acetylneuraminic acid (NeuAc) from CMP-NeuAc to lactose, {alpha}-methyl-galactopyranoside and β-methyl-galactopyranoside with low apparent Km and the {alpha}2,6-sialyltransferase catalysed the transfer of NeuAc from CMP-NeuAc to lactose with low apparent Km.

Key Words: bacterial sialyltransferase, glycosyltransferase family 80, photobacterium, sialic acid

Abbreviations: bis-Tris, 2-[bis(2-hydroxyethyl)amino]-2(hydroxymethyl)-1,3-propanediol; Bn, benzyl; bp, base pairs; CMP-NeuAc, cytidine monophosphate N-acetylneuraminic acid; Fuc, fucose; Gal, galactose; Glc, glucose; HPLC, high performance liquid chromatography; IPTG, isopropyl-1-thio-β-D-galactopyranoside; Me, methyl; [M-H]–, deprotonated molecular ion; MALDI-TOF MS, matrix-assisted laser desorption/ionization-time of flight mass spectrometry; NAc, N-acetyl; NaCl, sodium chloride; NeuAc, N-acetylneuraminic acid; NMR, nuclear magnetic resonance; PAGE, polyacrylamide gel electorophoresis; PCR, polymerase chain reaction; ph, phenyl


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.