This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Sakurada, K Articles by Hasegawa, M Search for Related Content PubMed PubMed Citation Articles by Sakurada, K Articles by Hasegawa, M

research-article

Cloning, expression, and characterization of the Micromonospora viridifaciens neuraminidase gene in Streptomyces lividans.

Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Japan.

ABSTRACT

We have cloned the Micromonospora viridifaciens neuraminidase (EC 3.2.1.18) gene (nedA) in Streptomyces lividans. This was accomplished by using the vector pIJ702 and BglII-BclI libraries of M. viridifaciens chromosomal inserts created in S. lividans. The libraries were screened for the expression of neuraminidase by monitoring the cleavage of the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-alpha-D-N-acetyl-neuraminic acid. Positive clones (BG6, BG7, BC4, and BC8) contained the identical 2-kb BclI-BglII fragment and expressed neuraminidase efficiently and constitutively using its own promoter in the heterologous host. From the nucleotide sequence analysis, an open reading frame of 1,941 bp which encodes a polypeptide with an M(r) of 68,840 was detected. The deduced amino acid sequence has five Asp boxes, -Ser-X-Asp-X-Gly-X-Thr-Trp, showing great similarity to other bacterial and viral neuraminidases. We have also identified the catalytic domain by using truncated proteins produced in S. lividans.

This article has been cited by other articles:

• Warwas, M. L., Watson, J. N., Bennet, A. J., Moore, M. M. (2007). Structure and role of sialic acids on the surface of Aspergillus fumigatus conidiospores. Glycobiology 17: 401-410 [Abstract] [Full Text]  
• Jermyn, W. S., Boyd, E. F. (2002). Characterization of a novel Vibrio pathogenicity island (VPI-2) encoding neuraminidase (nanH) among toxigenic Vibrio cholerae isolates. Microbiology 148: 3681-3693 [Abstract] [Full Text]  
• Jost, B. H., Songer, J. G., Billington, S. J. (2002). Identification of a Second Arcanobacterium pyogenes Neuraminidase and Involvement of Neuraminidase Activity in Host Cell Adhesion. Infect. Immun. 70: 1106-1112 [Abstract] [Full Text]  
• Jost, B. H., Songer, J. G., Billington, S. J. (2001). Cloning, Expression, and Characterization of a Neuraminidase Gene from Arcanobacterium pyogenes. Infect. Immun. 69: 4430-4437 [Abstract] [Full Text]  
• Mizan, S., Henk, A., Stallings, A., Maier, M., Lee, M. D. (2000). Cloning and Characterization of Sialidases with 2-6' and 2-3' Sialyl Lactose Specificity from Pasteurella multocida. J. Bacteriol. 182: 6874-6883 [Abstract] [Full Text]  
• Monti, E., Preti, A., Nesti, C., Ballabio, A., Borsani, G. (1999). Expression of a novel human sialidaseencoded by the NEU2 gene. Glycobiology 9: 1313-1321 [Abstract] [Full Text]  
• Chou, M.-Y., Li, S.-C., Li, Y.-T. (1996). Cloning and Expression of Sialidase L, a NeuAcalpha 2right-arrow 3Gal-specific Sialidase from the Leech, Macrobdella decora. J. Biol. Chem. 271: 19219-19224 [Abstract] [Full Text]