Barley malt endoproteases (EC.3.4.21) develop as multiple isoforms mainly during grain germination and pass through kilning almost intact. Thermostability, under simulated mashing conditions, varied from low to high depending on the substrate used in the assay. This suggests that individual enzymes respond differently to heat exposure and to protein substrates. The optimal pH with haemoglobin was pH 3.5, with hordein pH 4 and with glutelin pH 5. The optimal temperature with hordein was 40 o C, with glutelin 50 o C and with haemoglobin 60 o C. These differences suggest that it is not possible to comprehensively characterise all malt endoproteases under one set of assay conditions. In brewing, most of the barley protein degradation (> 70 %) occurs during malting. But some proteinases remain active during mashing and contribute to wort soluble proteins and free amino nitrogen. Their contribution to all malt EBC mash total free amino nitrogen was 25 % in Schooner (Australian) and 30 % in Morex (USA). The importance of proteolytic activity during mashing and the possibility that the levels may not be adequate, at high solid adjunct ratios, are acknowledged.

Barley and malt proteins, of infusion (IoB) and decoction (EBC) mashing worts as well as commercial wort and beer, obtained from the Castlemaine Perkins brewery, Brisbane, were gel filtered, with or without further treatments. A general, similar pattern of protein and peptide profiles emerged from barley malt and beer. This confirmed the widely assumed fact that beer proteins descend from barley, some transformed and others perhaps mostly unchanged by processing. In the gel-filtrate profiles, a maximum of 8 or 9 fractions were discerned. These fractions were collected and quantified for protein contents and amino acid compositions. The first four fractions contained the proteins and polypeptides of molecular weight higher than 14,000. Consequently, the remaining fractions contain the smaller peptides (<14,000), that were completely removed by dialysis. The effects of processing on proteins and peptides varied contingent upon the type of processing step considered and the pre-chromatographic treatment. Malting was the most effective process remarkably increasing the soluble protein contents, especially the smaller peptide fractions and the colour development. This is the first report, as far as we are aware of, on the gel filtration profiles of wort and beer low molecular weight peptides including those of barley wort. The importance of the smaller peptides in foam formation and retention cannot be overemphasised. The amino acid composition of the fractions revealed much more diversity than was observed in the comparison of the profiles. Proline content of fraction 1 resembled that of barley soluble proteins while fractions F2, F3 and F4 that of glutelin and only fraction 8 that of hordein. The latter, suggests that hordeins or, at least the peptide products rich in proline, are likely to be completely digested to amino acids, during malting.