Characterization of K-binding factor involved in water-soluble complex of menaquinone-7 produced by Bacillus subtilis natto

Abstract

Vitamin Ks are expected to contribute bone and cardiovascular health. Especially, menaquinone-7 has a higher bioavailability and a longer half-life than other vitamin Ks in the human body. However, their low water-solubility limits their application. On the other hand, Bacillus subtilis natto produces a water-soluble complex, which comprises menaquinone-7 and peptides. The peptide named K-binding factor (KBF) has been reported as the main component of the complex. In the present, the structural characteristics of KBF were studied. Mass spectrometry showed significant peaks at m/z = 1050, while the previous PAGE suggested that molecular weight of KBF was ~ 3k. Amino acid analysis revealed that the 1k peptides were the various combinations of nine amino acids, among which Asx, Glx, Val, Leu and Met were found to be the most abundant. The peptides could serve as detergent properties. The 1k peptides could be isolated by reverse-phase high performance liquid chromatography. The bundle of three 1k detergent-like peptides would participate to the micelle structure containing menqauinone-7 inside. In conclusion, a basic unit of KBF would be the ~ 1k peptides, and the three basic unit assemble to the ~ 3k bundle, then the bundle form a water-soluble micelle including menqauinone-7 inside.

Keywords: Bacillus subtilis natto; K-binding factor; menaquinone-7; solubilization; vitamin K.Abbreviations: CV, column volume; DLS, dynamic light scattering; FPLC, Fast protein liquid chromatography; KBF, K-binding factor; MK-7, menaquinone-7; natto-MK-7, a water-soluble complex of menaquinone-7; RH, hydrodynamic radius.