Quantitation by enzyme immunoassay of spirosin from Lactobacillus reuteri and Escherichia coli

Purification of spirosin from Lactobacillus reuteri DSM 20016 and partial characterization based on its amino acid sequences.

“…1, the reactivity with the MAb (i.e., spirosin) was eluted at 0.18 M KCl and ADH activity at 0.26 M. The two peaks were clearly distinct from each other. Together with the previous findings that three strains of L. reuteri (DSM 20015, 20016 and 20053) exhibited significantly different levels of ADH activity whereas they produced almost the same amounts of spirosin (Yamato et al, 1997), the chromatographic results demonstrated that the spirosin of L. reuteri was not ADH at all. In addition, another protein which had the same molecular weight of 95 kDa and therefore superimposed on spirosin on SDS-PAGE (Fig.…”

“…Affinity chromatography by use of the MAb was not successful either. Spirosin did not bind to the antibody ligand presumably because it might have a polymeric structure in a solution, and therefore its antigenic determinant might be buried as suggested previously (Yamato et al, 1997). It was finally obtained by a preparative SDS-PAGE as described in a previous paper (Yamato et al, 1994).…”