Alpha-amylase/ subtilisin inhibitor levels in Australian barleys

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Jarrett, SJ, Marschke, RJ, Symons, MH, Gibson, CE, Fox, GP & Henry, RJ 1997, 'Alpha-amylase/ subtilisin inhibitor levels in Australian barleys', Journal of Cereal Science, vol. 25, no. 3, pp. 261-266.

Journal of Cereal Science home page available at http://www.elsevier.com/locate/jcs
Publisher's version of article available at http://dx.doi.org/10.1006/jcrs.1996.0084

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An enzyme-linked immunosorbent assay (ELISA) was used to determine the bifunctional alpha-amylase/subtilisin inhibitor (BASI) content of barley grain from 11 cultivars grown in six diverse locations in Australia. The inhibitor ranged from 119 to 254 μg/g in 57 barley samples. Genotype had a significant (PPP<0·05) with alpha-amylase activity in corresponding malts. The ELISA used monoclonal and polyclonal antibodies raised against purified BASI. In immunoblot analysis the monoclonal antibody showed high specificity for the inhibitor in barley and also detected the inhibitor in wheat. Low levels of inhibitor (mean 3·2 μg/g) were found in 12 Australian wheat cultivars using the ELISA developed for barley. The assay had a linear working range of 5–50 ng/mL with a detection limit of 2 ng/mL. Reproducibility between assays was good (CV=4·9%) but mean recoveries were high, ranging from 116–129% when purified inhibitor was added to barley extracts. The ELISA may have useful applications in brewing research and barley breeding programmes.