Partial Purification of Ferulic Acid Esterase from Malted Barley
F.J. Humberstone 1, 2 and D.E. Briggs 1, 3, 4
1Birmingham Malting and Brewing Group, School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT
2Current address: Unipath Ltd, Bedford Priory Business Park, Bedford, MK44 3UP, UK
3Current address: 66 Sandhills Lane, Barnt Green, nr. Birmingham, B45 8NX, UK
4Corresponding author. E mail: d.e.briggs@bham.ac.uk
J. Inst. Brew. 108(4), 439-443, 2002 | VIEW ARTICLE
ABSTRACT
A partial purification of ferulic acid esterase, which degrades feruloyl glycerol, has been achieved from barley malt in small yields. Coloured and viscous materials were removed from the malt extract using batch-elution anion exchange chromatography. Further steps included gradient elution anion exchange chromatography and gel filtration chromatography. Estimations of the molecular weight varied greatly from 22KDa to 158 KDa, possibly because the protein interacted with the matrix of the gel exclusion chromatography column and because multiple forms of the enzyme were present. The partially purified feruloyl esterase had an apparent Km of 0.46% feruloyl glycerol. However more than one enzyme may be present and the substrate contains two isomers and so Michelis-Menten kinetics may not be appropriate.
Key words:
Barley malt, ferulic acid esterase, partial purification.
Publication no. G-2002-1230-001 ©2002 The Institute & Guild of Brewing