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(SE/BTI-Cme) and Its Influence on Beer Haze Stability

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(SE/BTI-Cme) and Its Influence on Beer Haze Stability ÊRSIII An investigation of a barley protein (SE/BTI'CMe) and its influence on beer haze stability by Louise H. Robinson B.Sc. (Hons), Deakin University A thesis submitted to the University of Adelaide for the degree of Doctor of Philosophy School of Agriculture and'Wine Waite Campus University of Adelaide South Australia November,20O4 I Declaration I hereby declare that this work does not contain any material previously submitted for another degree or diploma in any University or other tertiary institution. To the best of my knowledge and belief, it does not contain any material previously published or written, except where due reference is made in the text. I give consent to this copy of my thesis, when deposited in the University library, being available for loan and photocopying. Louise H. Robinson 11 Acknowledgements I wish to thank my principal supervisor Dr Evan Evans, along with my other supervisors Dr Chris Ford and Dr Andrew Barr for their supervision, guidance and critical reading of this thesis. In particular, special thanks must go to Evan for all his help, enthusiasm, technical advice and support as well as his encouragement of me to travel and see the world as part of my Ph.D. studies. I would also like to thank Dr Silja Home and Dr Anu Kaukovirta-Norja for all their support and enthusiasm. I would like to thank the members of the South Australian Barley Improvement Program field team for providing barley samples used in these experiments along with the Waite Barley Quality Evaluation Laboratory for the use of equipment and brewing analysis. To the members of my lab both past and present (Stewart, Helen, Judy, Nigel and Michael), thanks for the encouragement and friendship. I would like to thank Dr Silja Home for the kind invitation to complete four months of this project with her group at VTT Biotechnology. Thanks must go to Mr Arvi Vilpola and Dr Anu Kaukovirta-Norja for all their assistance and advice in completing a large number of pilot brewing trials in a very short space of time. I would like to thank everyone at VTT Biotechnology who made my stay in Finland most enjoyable. Eero 'Wikström, Mattila, Vuokko Liukkonen, Sirkka Kanervo, Tarja Marke Holm, and Hannele Sweins are to be thanked for their technical assistance with the brewing trials. I would like to thank Dr Peter Aldred for his assistance and advice with pilot brewing trials, as well as the University of Ballarat for the provision of pilot brewing facilities. lll I would like to thank Tooheys Brewery and the South Australian Brewing Company for the use of equipment, assistance with brewing trials and brewing analysis. Thanks also must go to AusMalt, Kirin Australia, Polttimo Companies, Raisio Malt, Barrett Burston Malting Company and the International Malting Company for the provision of malt samples. I would like to thank Mr Jelle Lahnstein for his assistance and advice with the SE protein analysis and Dr Andrew Milligan and Dr Juan Juttner for their assistance and advice on the cloning and purification of BTI-CMe. I would like to thank Mr Lance Lusk and Dr David Ryder for inviting me to participate in the Miller brewing companies summer internship program in the suÍìmer of 2003. A big thanks to everyone at Miller who made my stay in Milwaukee most enjoyable. I wish to acknowledge the financial support of the Australian Research Council, AusMalt, Lion Nathan, VTT Biotechnology, the Finnish malting and brewing industry, and the Grains Research and Development Corporation. Thank you to all my family and friends for their support and encouragement during the course of my studies. Finally, I would like to acknowledge my fiancé Dave, thank you for your unconditional love, support and encouragement, thanks for everything. This thesis is dedicated in memory of my mother, Marie Robinson 194I-2001 1V List of Abbreviations and Acronyms ü, B beta v Vo 2-D two-dimensional AAL attenuation limit AFLP bp base BASI lase/subtilisin inhibitor BBBI Bowman-Birk inhibitor BSA bovine serum albumin BTI bar inhibitor BU bitterness units "C CHAPS cM centimorgan cm centimetre CMe proteins chloroform-methanol soluble proteins cv cultivar DE diatomaceous earth DH double haploid dNTP 2'deoxynucleoside 5'-triphosphate DNA acid DP diastatic DTT dithiothreitol EBC Convention EDTA lenediaminetetraacetic acid ELISA linked immunosorbent as EMBL Molecular EtOH ethanol F.A.N. Free Amino Ni FU formazin units o gram h hour HRP horseradish peroxidase HWE hot water extract IEF isoelectric focusing IPTG isopropyl kb kilobase kDa kilodalton KI Kolbach Index KJ kilo Joule L litre LB Luria bertani LSD least difference LTP transfer M molar v MM millimolar MAS marker assisted selection mg millieram mln minute mL millilitre mg,lg millierams per gram mglI' millisrams per litre me/mL millisrams per millilitre mm millimetre mPa.s millipascal second MW molecular weight N nitrogen NCBI National Centre for Biotechnology Information ng nanogram Ni-NTA Nitrilotriacetic acid nm nanometre nmole nanomole NTU nephelos turbidity units o oxygen OD optical Density "P degrees Plato PAGE lamide PBS buffered saline PCR polymerase chain reaction pI isoelectric point pmol picomole ppm parts per million PT protein/tannin PTH phenylthiohydantoin PU pasteurisation units PVPP polyvinylpolypyrolidone QTL quantitative trait locr RFLP restriction fragment length polymorphism RNase A ribonuclease RP-HPLC reversed phase-high pressure liquid chromatography rpm revolutions per minute sarkosyl N-lauroylsarcosine SE silica eluent sec second SDS sodium dodecyl sulphate SG specific gravity SP soluble protein SNPs nucleotide SSB small scale brewing SSR simple sequence repeats TA tannic acid TAE Tris-Acetate-EDTA Taq thermus aquaticus DNA Polymerase TBS tris buffered saline VI TE TrisÆDTA TEMED N -Tetra- I TFA trifluoroacetic acid TIA inhibitor TM trade mark TPBS tween buffed saline TP total protein Tris-HCl amino methane Tween 20 polyoxyethylenesorbitan monolaurate t[L microlitre Lls ws.lmI' millilitre pm micrometre U units UV ultraviolet V volt Vh volt hour vlv volume to volume ratio w watt WK Windisch-Kolbach w/v to volume ratio X times vrl Publications arising from this thesis Robinson, L.H., Evans, D.E., Kaukovirta-Norja, 4., Vilpola, 4., Aldred, P., and Home, S. (2004). The interaction between malt protein quality and brewing conditions and their impact on beer colloidal stability. Tech, Q. Master Brew. Assoc. Am., 4l:353-362. A copy of this paper is included at the back of this thesis. Robinson, L.H., Ford, C.F., Healy, P., Gibson, C.E., Barr, A.R., and Evans, D.E' (2004). The genetic basis of improved beer haze stability. Proc. International Barley Genetics Symp., 9th, Brno, pp. 493-498. nEvans, D.E., Robinson, L.H., Sheehan, M.C., Tolhurst, R.L., Hill, 4., Skerritt, J.S., and Barr, A.R. (2003). Application of immunological methods to differentiate between foam-positive and haze active proteins originating from malt. 'L Am. Soc. Brew. Chem.,6l(2):55-62. A copy of this paper is included at the back of this thesis. 'This publication does not constitute a major part of the work presented in this thesis. Robinson, L.H., Ford, C.F., Healy, P., Gibson, C'8., Barr, 4.R., and Evans, D.E. The genetic basis of a malt haze active protein. In preparation. Robinson, L.H., Ford, C.F., Healy, P., Gibson, C.E., Barr, 4.R., and Evans, D.E' Identification of a barley SE/BTI-CMe protein that influences beer haze stability. J. Cereal Sci. In preparation. Robinson, L., vilpola, 4., Kaukovirta-Norjâ, 4., Evans, E., and Home, S. (2003). The interaction between malt protein quality and brewing conditions and their impact on beer colloidal stability. Proc. Master Brew. Assoc. Am., Convention, Milwaukee, WL, Oral II (CD-ROM). vlll Robinson, L.H., Home, S., Pöyri, S., Aldred, P., Healy, P., Gooden, J', Barr, A' R', and Evans, D. E. (2003). The influence of the barley trypsin inhibitor CMe (BTI-CMe) on the potential colloidal stability of beer. In Proc. I lth Aust. Barley Tech. Symp', Adelaide, pp. 250-253. Robinson, L., Home, H., Pöyri, s., Aldred, P., Healy, P., Gooden, J., Barr, 4., and Evans, E. (2003). Influence of the barley trypsin inhibitor CMe (BTI-CMe) on the potential colloidal stability of beer. Am. Soc. Brew. Chem', Convention, Santa Ana Pueblo, NM, poster #5 http://www.asbcnet.org/ meetin gs/2003/abstracts/p- 5.htm Robinson, L., Sheehan, M., Healy, P., Gooden, J., Barr, A. R., Ford, C. M', and Evans, D. E. (2002). The influence of malt quality on the colloidal (haze) stability of beer during storage and transport. In: Proc. 2lh Conv. Inst. Brew. Asia- Pacific section, Adelaide, poster #5 (CD-Rom)' Robinson, L., Sheehan, M., Stuart, I. M., Barr, A. R., Ford, C. M., and Evans, D. E. (2001). The influence of malt quality on the colloidal stability of beer. In: Proc. 10th Aust. Barley Tech. Symp., Canberra, poster #30 (CD-Rom). 3b0f8 109 e23 c2f 4 c e8 c ade c6641 22#T opOfP age 1X Table of Contents Title Page I Declaration ii Acknowledgements iii List of Abbreviations and Acronyms v Publications arising from this thesis viii Table of Contents 1 Abstract 7 Chapter L - Literature Review 10 1.1 Introduction 10 l.2The brewing process and its impact on the potential haze stability of beer 11 1.2.1 Raw materials 12 l.2.Il Malting 12 l.2.2Brewing 13 1.2.3 Fermentation and Maturation 14 l.2. Ftltration and Packaging 15 1.2.5 Storage and Transport 15 l.2.6Beer Styles 16 1.3 Protein-polyphenolhaze measurement I7 l. Praze Active (HA) proteins 18 1.4.1 Hordein 20 1.5 HA polyphenols 23 1.6 Protein:Polyphenol interactions in colloidalhaze formation
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