Fungal hydrophobins have been shown to induce gushing of beer. In order to study the occurrence and fate of hydrophobins at different stages of the production chain of beer, barley samples artificially infected in the field with Fusarium culmorum, F. graminearum and F. poae were collected during the growing period as well as during various stages of the malting process. In addition, naturally infected malt was brewed in pilot scale and samples were collected throughout the process. The samples were assayed for hydrophobin content using an ELISA method. The results showed that fungi produced hydrophobins that accumulated during barley grain development in the field, but that production was more pronounced during malting. Prolonged storage of barley tended to reduce the ability of fungi to produce hydrophobins in malting. Studies on the fate of hydrophobins during the brewing process revealed that mashing released hydrophobins from the malt into the wort. Some loss of hydrophobins occurred throughout the brewing process with spent grains, cold break (wort boiling) and surplus yeast. In addition, the beer filtration step reduced hydrophobin levels. Despite the substantial loss of hydrophobins during brewing, the level was high enough to induce the gushing detected in the final beer.

Lactobacillus plantarum VTT E-78076 (E76) and Pediococcus pentosaceus VTT E-90390 (E390) starter cultures were added to the steeping water of normal malting barley in order to balance the microbial community and to enhance malt processability. In this study, we also investigated the effects of lactic acid-acidified MRS-spent medium (MRS-LA) on malting performance. Malting trials with five different two-row barley varieties were carried out in 25 kg pilot scale. The starter cultures promoted yeast growth during malting and restricted the growth of harmful bacteria and Fusarium fungi. Furthermore, they had positive effects on malt characteristics. Reduction in wort viscosity and beta-glucan content and enhanced xylanase and microbial beta-glucanase activities were observed. Starter cultures notably improved lautering performance. Some of the beneficial effects were due to the lactic acid and low pH, as similar effects were obtained with MRS-LA. Starter cultures offer a tool for tailoring of malt properties.

The optimal pitching rate in high gravity worts (12-16°P) was about 0.3 g/l wet weight (2.3 x 106 counted cells/ml) and per one percent of original wort gravity. In very high gravity worts (20-23°P) the corresponding figure was 0.4 g/l (2.9 x 10* cells/ml). Higher amounts of yeast did not improve the fermentation rate.With increased original wort gravity, flocculation of the yeast weakened and the amount of cropped yeast decreased. The viability of the crop yeast was good.In the conditions used, excessive production of acetate esters occurred only with pitching rates lower than the recommended rate. As the original wort gravity increased, more fermentable extract was metabolized to ethanol rather than utilized for yeast growth. The highest ethanol yield obtained was 10.9% (v/v).

Four different types of endo-fl-l,4-glucanase active bottom-fermenting brewer's yeast strains were constructed using recombinant DNA technology. To study the effects of different promoters, copy numbers and integration sites, the eoll gene of the filamentous fungus Trichoderma reesei was inserted between the promoter and terminator regions of either the PGK1 or ADH1 gene of yeast. The eoll gene was transferred to the industrial brewer's yeast on a multicopy plasmid or alternatively integrated into the LEU2, PGK1 or A D H 1 locus of the yeast. Integration into the PGK1 or A D H 1 locus did not affect the brewing properties of the yeast or the quality of the finished beer. Integration into the LEU2 locus, however, decreased the metabolic activity of yeast and prolonged fermentation was needed. In pilot brewing conditions the PGK1 promoter was stronger than that of ADH1. Even a single copy of the eoH gene in the PGK1 integrant strains gave rise to sufficient enzyme activity for the hydrolysis even of unusually high total amounts of fl-glucans in worts.

The steep water is generally aerated in industrial barley malting. However, it is questionable whether oxygen actually reaches the embryo, which remains entrapped under the husk, testa, and pericarp until chitting occurs. The aim of our study was to investigate whether barley embryos experience oxygen deficiency during steeping, and whether various steeping conditions affect the oxygen deficiency. Alcohol dehydrogenase Adh2 was induced in all steeping conditions studied. Therefore, oxygen deficiency occurred regardless of the steeping conditions. However, steeping conditions affected the rate of recovery from oxygen deficiency, germination rate, and onset of alpha-amylase production. When barley was subjected to oxygen deficiency by applying N(2) gas during steeping, the timing of the treatment determined its effects. The importance of aeration increased as the process proceeded. Oxygen deprivation at the beginning of the process had little effect on malt quality. Therefore, the timing of aeration is important in the optimization of germination during the steeping stage of malting.