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Identification and functional analysis of fructosyl amino acid‐binding protein from G ram‐positive bacterium A rthrobacter sp.
Author(s) -
SakaguchiMikami A.,
Ferri S.,
Katayama S.,
Tsugawa W.,
Sode K.
Publication year - 2013
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12152
Subject(s) - identification (biology) , bacteria , arthrobacter , gram , microbiology and biotechnology , biology , gram negative bacteria , bacterial protein , biochemistry , computational biology , escherichia coli , genetics , gene , ecology
Abstract Aim Fructosyl amino acid‐binding protein ( FABP ) is a substrate‐binding protein ( SBP ), which recognizes fructosyl amino acids ( FA s) 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. FV 1‐1, 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 SBP s 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 N α‐ FA ‐specific binding activity by intrinsic fluorescent measurement. Conclusion This study identified a novel FABP from a Gram‐positive bacterium, Arthrobacter sp., which exhibited N α‐ 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 N α‐ 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 SBP s 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 N α‐ FA ‐sensing molecules.