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The identification of a barley haze active protein that influences beer haze stability: Cloning and characterisation of the barley SE protein as a barley trypsin inhibitor of the chloroform/methanol type

journal contribution
posted on 2023-05-16, 17:06 authored by Robinson, LH, Juttner, J, Milligan, A, Lahnstein, J, Eglinton, JK, Evans, E
Previous pilot brewing trials have demonstrated that in the absence of a molecular weight (MW) ~12,000 (barley silica eluate (SE) protein), the beer brewed from the malt of these SE -ve varieties formed less haze after accelerated ageing than beers brewed using SE +ve malt varieties. The previously described SE protein was characterised using comparative two-dimensional (2-D) gel electrophoresis immunoblots of barley seed extracts from both SE +ve and SE -ve varieties. The SE protein spot identified was excised and its partial sequence determined, after in-gel cleavage using trypsin and separation of the resulting fragments by reversed-phase high performance liquid chromatography (HPLC). N-terminal sequence analysis of the tryptic peptides from SE +ve and SE -ve varieties identified the SE protein as the barley trypsin inhibitor-CMe precursor (BTI-CMe). The mature BTI-CMe protein is 13.3 kDa and its functional gene is located on chromosome 3H. Cloning of the BTI-CMe protein demonstrated that both SE -ve and SE +ve barley varieties contain a BTI-CMe protein family member that is similar but has a consistently different sequence, primarily in the last 30 amino acid residues of their C-termini.

Funding

Australian Research Council

AusMalt Pty Ltd

History

Publication title

Journal of Cereal Science

Volume

45

Pagination

343-352

ISSN

0733-5210

Department/School

Tasmanian Institute of Agriculture (TIA)

Publisher

Academic Press

Place of publication

United Kingdom

Repository Status

  • Restricted

Socio-economic Objectives

Barley

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    University Of Tasmania

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