Advanced Method for Measuring Proteinase A in Beer and Application to Brewing*

Proteinase A, excreted from yeast cells into beer during fermentation in the brewing process, has been shown to degrade foam-active proteins and to decrease foam stability. In order to improve the measurement of this enzyme in beer, a new fluorescent peptide, MOCAc-Ala-Pro-Ala-Lys-Phe-Phe-Arg-Leu-Lys (Dnp)-NH₂, was synthesised and applied to the accurate and rapid estimation of proteinase A in commercial beer and fermenting wort. This novel substrate is several hundred times more sensitive to proteinase A than other previously reported synthetic substrates or native protein substrates. The concentration of proteinase A in beer is closely related to foam stability and proteinase A activity was found to increase gradually during fermentation. The concentration of proteinase A excreted from yeast cells is also closely related to the vitality of pitching yeast cells. This new method was successfully applied to the evaluation of yeast vitality and the development of optimum yeast handling procedures.     

姜汁啤酒的研制

针对冬季啤酒市场,开发姜汁保健啤酒。采用生姜渣汁分离,再对姜汁进行处理,姜渣在糖化工段加入,姜汁在发酵工段加入,酿制出的产品质量好,口味独特,适应冬季饮用,具有防病健身,理气健胃的保健作用。     

影响啤酒风味物质简述

影响啤酒风味的物质可分为醇、酯、碳基化合物、酸、含硫化合物、胺（挥发性）和酚基化合物等。产生这种物质主要来源是麦芽、谷物辅料、酒花、酵母的发酵等。主要阐述了由于发酵作用而产生的影响啤酒风味的形成机理。双乙酰也是影响酒风味的重要原因之一。     

THE INFLUENCE OF ENTEROBACTER AGGLOMERANS ON BEER FLAVOUR

The influence of Enterobacter agglomerans (Erwinia herbicola) on the fermentation process and beer flavour was studied. The presence of E. agglomerans gave rise to increased levels of acetaldehyde, methyl acetate, diacetyl, 2,3-pentanedione and dimethyl sulphide in the final product. The concentration of these compounds was affected by the number of bacterial cells inoculated into the pitched wort.     

2μm DNA PLASMID IN BREWERY YEASTS

Using an agarose gel screening procedure, 2 μm DNA plasmid was detected in all of 10 brewing strains of Saccharomyces yeast examined and in both of two non-brewing, dextrin-utilising strains. Plasmid DNA was identified in yeast grown with access to air in MYGP medium or in hopped wort, and in yeast harvested from 3-day wort fermentations. The yeast plasmid is a suitable self-cloning vector for the genetic manipulation of brewing yeasts by transformation.     

MITOCHONDRIAL RELEVANCE TO YEAST FERMENTATIVE PERFORMANCE: A REVIEW

It is well known that dissolved oxygen fulfils critical roles in brewing yeast physiology and overall fermentative performance. The major and minor roles that have been identified are briefly discussed and another role, that of providing for minimal mitochondrial development and functionality, is suggested. The long accepted theory that mitochondria are irrelevant to fermentative performance is reviewed as to its basis and the evidence in support of it. However, minimal mitochondrial development is required to provide the cell with critical metabolic intermediates and components. These are identified and reviewed and finally, evidence is presented that mitochondria are critical to brewing yeast fermentative performance. The review concludes that when assessing the role of mitochondria, concern should be broader than simply for the energetic function of these organelles.     

DETERMINATION OF HIGH MOLECULAR WEIGHT β-GLUCAN IN WORT AND BEER USING A POST-COLUMN CALCOFLUOR FLOW-INJECTION-ANALYSIS (FIA) METHOD

To eliminate the influence of maltose, ethanol, low molecular weight β-glucan and an inhibitor of the calcofluor fluorescence reaction in wort and beer on the measured values of a calcofluor-FIA, a post-column calcofluor-FIA method has been developed using a short-size gel permeation chromatography column (6.0 × 50 mm). A column packed with polyhydroxymethacrylate gel (molecular weight exclusion limit, 100,000) was found to be the most appropriate for this system. This short column allowed rapid and specific measurement of high molecular weight β-glucan in a few minutes without the influence of the fluorescence inhibitor, maltose and ethanol which have molecular weights of less than 1000 daltons. Because the low molecular weight β-glucan responsible for the scatter caused by a slight difference in measuring conditions such as temperature, calcofluor concentrations, sample volumes, etc., was separated through the use of the column, the measured values by the post-column calcofluor-FIA method hardly fluctuated under different conditions. Though it has been recognized that dilution of a sample could affect a calcofluor FIA, the new system was not influenced. This also made it possible to measure the β-glucan content in dark-coloured samples (even over 100 EBC colour units). The measured values by the post-column FIA showed a high correlation (r² = 0.993) with those obtained by the enzymatic method (the McCleary method).     

APPLICATION OF THE POLYMERASE CHAIN REACTION TO THE RAPID ANALYSIS OF BREWERY YEAST STRAINS

The rapid discrimination of closely-related Saccharomyces cerevisiae strains can pose a significant problem to breweries, in particular where closely related strains are being used simultaneously to manufacture different products. In this study, two PCR approaches have been examined to assess their usefulness for the discrimination of brewery ale and lager yeast strains. PCR using arbitrary primers (RAPD PCR) was found unsuitable for such an application since the DNA profiles generated from brewery strains were generally found to be identical, due presumably to the close genetic relatedness of these yeasts. In contrast, PCR using δ sequence primers could rapidly differentiate between many ale and lager strains and characteristic profiles for these were generated. This method could also be applied directly to yeasts isolated from brewery worts or from active dried yeast preparations. Results of such analyses were available within the working day.     

外加酶法酿制低糖啤酒糖化工艺的研究

以普鲁兰酶Ｐｒｏｍｏｚｙｍｅ１２０Ｌ为重点，综合分析了外加酶糖化过程中影响麦汁总还原糖量和糖组成的各种因素，如各酶制剂的用量、糖化温度、料水比、物料比及各因素之间的相互作用等，确定了一套最优的糖化工艺方案。所得麦芽汁浸出率高、色度浅、粘度低、还原糖含量高；经过高效液相色谱分析（ＨＰＬＣ）表明，其糖组成合理；经过十天的发酵，发酵度达８２．２％，酒精分为６．４５５％（Ｗ／Ｗ）。