Abstract
Bacillus subtilis is one of the few strains that can secrete synthetic menaquinone-7 (MK-7) to the outside of the cell, and its purpose and mechanism have not been clearly studied. As an amphiphilic protein naturally synthesized by Bacillus subtilis, the BslA protein may be involved in the inversion of extracellular vitamin K2 solubility. The protein structure in UniProt was used to search for the possible binding sites of MK-7, and the analysis of the higher ranking results of the genetic algorithm showed that the ASP166 residue was likely to be the binding site. They could form a stable hydrogen bond connection through ASP166, and approximately 7 proteins formed the conformation of a fixed naphthoquinone ring. We isolated and obtained the BslA protein by Ni-NTA affinity chromatography. Then, MK-7 was modified by BslA in vitro. A series of experiments, such as SEM, XPS, and WCA, showed that MK-7 and BslA proteins can realize self-assembly and transform from fat-soluble to water-soluble complexes. When the bslA protein in Bacillus subtilis natto was overexpressed, its MK-7 synthesis ability was further improved, especially the extracellular MK-7 content, which increased by 16%. This finding suggests that the BslA protein in Bacillus subtilis is likely to be involved in the extracellular secretion of MK-7 as a receptor.
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