Identification and functional analysis of fructosyl amino acid-binding protein from Gram-positive bacterium Arthrobacter sp.
Article first published online: 18 FEB 2013
© 2013 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 114, Issue 5, pages 1449–1456, May 2013
How to Cite
Sakaguchi-Mikami, A., Ferri, S., Katayama, S., Tsugawa, W. and Sode, K. (2013), Identification and functional analysis of fructosyl amino acid-binding protein from Gram-positive bacterium Arthrobacter sp. Journal of Applied Microbiology, 114: 1449–1456. doi: 10.1111/jam.12152
- Issue published online: 15 APR 2013
- Article first published online: 18 FEB 2013
- Accepted manuscript online: 30 JAN 2013 04:30AM EST
- Manuscript Accepted: 20 JAN 2013
- Manuscript Revised: 26 DEC 2012
- Manuscript Received: 30 SEP 2012
- Arthrobacter ;
- fructosyl amino acid-binding protein;
- fructosyl amino acid;
- Gram-positive bacteria;
- Substrate-binding proteins
Fructosyl amino acid-binding protein (FABP) is a substrate-binding protein (SBP), which recognizes fructosyl amino acids (FAs) as its ligands. Although FABP has been shown as a molecular recognition tool of biosensing for glycated proteins, the availability of FABP is still limited and no FABP was reported from Gram-positive bacteria. In this study, a novel FABP from Gram-positive bacteria, Arthrobacter spp., was reported.
Method and Results
BLAST analysis revealed that FABP homologues exist in some of Arthrobacter species genomes. An FABP homologue cloned from Arthrobacter sp. FV1-1, FvcA, contained a putative lipoprotein signal sequence, suggesting that it is a lipoprotein anchored to the bacterial cytoplasmic membrane, which is a typical characteristic for SBPs from Gram-positive bacteria. In contrast, FvcA also exhibits high amino acid sequence similarity to a known Gram-negative bacterial FABP, which exists as a free periplasmic protein. FvcA, without the N-terminal anchoring region, was then recombinantly produced as soluble protein and was found to exhibit Nα-FA-specific binding activity by intrinsic fluorescent measurement.
This study identified a novel FABP from a Gram-positive bacterium, Arthrobacter sp., which exhibited Nα-FA-specific binding ability. This is the first report concerning an FABP from a Gram-positive bacterium, suggesting that FABP-dependent FA catabolism system is also present in Gram-positive bacteria.
Significance and Impact of the Study
The novel FABP exhibits the ability to specifically bind to Nα-FA with a high affinity. This selectivity is beneficial for applying FABP in HbA1c sensing. The successful preparation of water-soluble, functionally expressed Gram-negative bacterial FABP may make way for future applications for a variety of SBPs from Gram-positive bacteria employing the same expression strategy. The results obtained here enhance our understanding of bacterial FA catabolism and contribute to the improved development of FABP as Nα-FA-sensing molecules.