| Abstract: | Grape seed procyanidins were fractionated through different degrees of polymerisation, and human saliva was purified and separated into two fractions: one was mostly α‐amylase and the other was essentially proline‐rich proteins (PRPs). The interaction of these proteins with the procyanidin compounds was assayed using nephelometry, and the influence of several factors was investigated, such as degree of polymerisation, pH and concentrations of both protein and tannin. The same experiments were performed with bovine serum albumin (BSA). The amount of insoluble aggregates, resulting from the formation of polyphenol–protein aggregates, increased quickly up to a maximum value which thereafter remained practically unchanged. pH was set at 5.0 for all further assays, since it was the nearest value to that encountered in human saliva (pH 5.6–7.9), where proteins were stable and had a maximum ability to bind and precipitate procyanidin oligomers. These proteins were shown to have a strong affinity for procyanidin oligomers and were unable to resolubilise the polyphenol–protein aggregates when present in excess. PRPs required a much lower content to bind all the tannins (400 µg of procyanidin oligomers) than BSA and especially α‐amylase (48, 60 and 132 µg respectively). The procyanidin's ability to bind PRPs, BSA and α‐amylase increased with its average molecular weight. This ability increased regularly for PRPs up to 4500 Da, whereas the ability to bind the globular proteins decreased beyond 3400 Da. © 2001 Society of Chemical Industry |
