THE INFLUENCE OF MASHING CONDITIONS ON WORT AND BEER QUALITY

From a comparison of the performance of ale and lager malts mashed by constant temperature infusion, by temperature programmed infusion, and by decoction procedures it emerges that malt modification and the pattern of kilning are as important in determining the composition of wort as are the mashing conditions. Worts with appropriate levels of amino and total nitrogen and of fermentable sugars may be obtained by selecting lightly-dried well-modified malts and mashing with a constant temperature infusion rather than by using less modified malt in conjunction with more complex mashing programmes. Ale malts yield worts of lower pH which is reflected in a slight reduction in hop utilization. Head retention is improved by the use of undermodified malts but colloidal stability is improved when well-modified malts are used. In the case of well-modified malts a high final curing temperature is not a prerequisite for achieving good colloidal stability in beer.     

CHELATORS AND METAL BUFFERING IN FERMENTATION

Trace elements are important components of brewer's wort. The effect of these metals is dependent upon the metal binders, (chelators) present. These chelators are wort components with reactive groups such as carboxyl, sulphydryl, o-diphenol or amine. The properties of different types of complex are briefly reviewed together with the general equations for their stability. The stability of some of the complexes formed in wort is comparable with the stability of EDTA complexes. The distribution of metals can be calculated by means of the equation for complex formation. This is the distribution between free ionized form and chelated forms. The ratio between free and bound forms or between assimilable and not assimilable complexes can be regulated by addition of EDTA. The calculated levels of free metals are compared with the formation of isobutanol in experimental fermentations. The same low free metal levels obtained with EDTA in a synthetic medium may be present in an all malt wort.     

PRODUCTION OF BEER USING IMMOBILIZED YEAST ENCODING α-ACETOLACTATE DECARBOXYLASE

Genetically modified brewer's yeast encoding α-acetolactate decarboxylase (α-ALDC) was tested in immobilized yeast bioreactors for main fermentation of beer. The α-ALDC enzyme produced by the transformant catalyzes the direct conversion of α-acetolactate to acetoin without formation of diacetyl. The long lagering period required for beer maturation in conventional brewing can thus be shortened or even omitted. Three different packed bed bioreactors were employed, with volumes of 1.6 dm³, 5 dm³ and 25 dm³. The 5 dm³ column had a slightly conical geometry in contrast to the others which had cylindrical shapes. Sintered glass beads were chosen as the carrier material on the basis of experiments with the parent strain. The brewing performance of the transformant compared well with that of the parent strain in the immobilized system. Fermentation, utilization of amino acids (including isoleucine, valine and leucine) and flavour formation were practically identical with both strains, the only difference being a marked decrease in the formation of diacetyl by the transformant. Small differences were, however, observed in the long-term biochemical stability. By using yeast encoding α-ALDC in the immobilized yeast system the total (primary and secondary) fermentation time could be reduced to approximately 2–6 days, compared with 3–6 weeks in a conventional batch process.     

RAPID BEER PRODUCTION AND CONDITIONING USING A PLUG FERMENTOR

Beer has been produced very rapidly, with residence time of less than 1 hour, by passing wort continuously through a plug formed from a yeast/kieselguhr mixture. Using malt wort the total output from the fermentor is limited by gradual blockage of the plug; nevertheless output is high per unit of yeast used. Prolonged operation is possible using nitrogen-free sugar solutions. The properties of beer produced from normal wort vary over the period of operation of the fermentor and blending is required to give a constant product. The average pH and nitrogen content of the beer is high if conventional wort is used but can be reduced to normal levels by control of wort composition. Details of a procedure for controlling the content of diacetyl and 2,3-pentane-dione in the beer are described and the potential value of a yeast-plug unit for beer conditioning is discussed.     

α-GLUCOSIDASE, α-GLUCOSIDE PERMEASE,MALTOSE FERMENTATION AND LEAVENING ABILITY OF BAKER'S YEAST

α-Glucosidase and α-glucoside permease are synthesized simultaneously in two different strains of baker's yeast (Saccharomyces cerevisiae) when the cells are induced with maltose. α-Thioethyl D-glucopyranoside (α-TEG) inhibits the transport of maltose into the cells, and the transport of maltose and α-TEG appeared to be mediated by the same permease system. There is an obvious correlation between α-glucosidase, α-glucoside permease and maltose fermentation activities in the yeasts while no correlation between these and the leavening ability of the yeasts can be demonstrated. Apparently the glucose concentration in dough is high enough to inhibit the permease-mediated transport of maltose into the cells thus impairing leavening ability from the maltose fermenting system.     

SUPPRESSION OF α-ACETOLACTATE FORMATION IN BREWING WITH IMMOBILIZED YEAST

Immobilized yeast cells extensively produced the diacetyl precursor, α-acetolactate, during alcohol fermentation. The activity of acetohydroxy acid synthetase, which is responsible for the formation of α-acetolactate from pyruvic acid, was high in cell-free extracts of immobilized yeast cells compared with that of free yeast cells. It was suggested that the expression of AHA synthase of immobilized yeast cells was increased during growth in the carrier as compared with free yeast cells. When the initial immobilizing yeast cell concentration was changed from 1.0 × 10⁶ cells/ml to 1.0 × 10⁹ cells/ml, production of α-acetolactate was reduced from 0.94 mg/l to 0.30 mg/l. Furthermore, during continuous fermentation for 10 d, the concentration of α-acetolactate in beer was 0.30 mg/l.     

A NOVEL AND PRACTICAL YEAST VITALITY METHOD BASED ON MAGNESIUM ION RELEASE

Using a commercial lager brewing yeast, the immediate release of magnesium, potassium and phosphate ions by cells when inoculated into wort was evaluated to be directly related to its subsequent fermentation performance. Yeast which released appreciable amounts of these ions immediately after inoculation mediated improved fermentations as evidenced by better growth, higher ethanol concentrations and lower diacetyl levels at the end of fermentation. Conversely, yeast that absorbed these ions or released them at very low concentrations performed poorly throughout fermentation producing beers with lower ethanol concentrations and higher diacetyl levels. These observations led to the identification and development of a rapid, practical and highly sensitive method to measure Mg⁺⁺ released or absorbed by yeast as an indicator of its vitality and a predictor of its subsequent fermentative performance. Full method details of the Magnesium Release Test (MRT) are given.     

发酵大麦提取物对3T3-L1前脂肪细胞分化及脂代谢的影响

研究乳酸菌发酵大麦提取物(LFBE)对3T3-L1前脂肪细胞分化及其脂代谢的影响与作用途径。利用植物乳杆菌(Lactobacillus plantarum dy-1)发酵大麦获得LFBE,采用Folin-Ciocalteu法测定总酚含量;凯氏定氮法测定蛋白质含量与苯酚硫酸法测定多糖含量;采用CCK-8法检测细胞增殖能力和油红O染色法分析前脂肪细胞的分化程度;采用Realtime PCR法检测LFBE对脂代谢关键基因转录水平的调控作用。结果表明,与未发酵大麦提取物相比,LFBE中总酚含量增加了20.88%;LFBE中蛋白质含量从发酵前的13.93%增加至34.94%;多糖含量从未发酵时的64.94%下降至35.43%。与未发酵大麦提取物组相比,LFBE能够有效地抑制3T3-L1前脂肪细胞的生长,其IC_(50)约为560μg/mL;与模型组和未发酵大麦提取物组相比,400μg/mL LFBE能够显著抑制3T3-L1前脂肪细胞分化,抑制率达到58.85%。与模型组相比,LFBE能显著抑制脂代谢关键基因C/EBPα、PPAR-γ、SREBP-1c、PTP1B和aP2的mRNA表达水平,上调GLUT4的mRNA表达水平(p0.05)。LFBE可有效抑制3T3-L1前脂肪细胞的增殖,并通过调节脂代谢相关基因的转录水平抑制其分化,表明其可能具有预防肥胖的作用。     

生物催化合成1,3-丙二醇工艺的研究

以甘油为底物,利用克雷伯氏菌发酵生产1,3-丙二醇的实验中,考察了初始甘油浓度、温度、pH、通气策略等发酵条件对1,3-PDO产量的影响。实验结果表明：积累1,3-PDO的适合条件为：甘油的初始浓度为40g/L、发酵温度37℃、pH7.0、0.5V/V·min的通气量,发酵30h,反应液中PDO的产量可达57.63g/L。     

SUGAR UTILIZATION BY A BREWING YEAST IN RELATION TO THE GROWTH AND MAINTENANCE PHASES OF METABOLISM

In batch fermentations, carried out using a range of conditions, the specific rate of sugar utilization is shown to decline throughout. When yeast mass is increasing, the yield coefficient relating carbohydrate utilization to mass production remains approximately constant and the declining specific fermentation rate parallels declining specific growth rate. When mass production ceases, maintenance of the yeast population requires only a low rate of sugar utilization. Attenuation associated with maintenance phase activity is relatively insignificant compared with that associated with growth.     

水相体系中固定化Acetobacter sp.CCTCC M209061细胞催化(S)-3-氯苯丙醇不对称合成

光学纯的3-氯苯丙醇是合成抗抑郁类药物的重要中间体。本论文报道了水相体系中固定化Acetobacter sp.CCTCC M209061细胞高效、高选择性地催化3-氯苯丙酮不对称还原为(S)-3-氯苯丙醇,采用聚乙烯醇与海藻酸钠的混合载体对醋酸杆菌细胞进行固定化,所得的固定化细胞的稳定性(热稳定性、p H稳定性及操作稳定性)均明显优于游离细胞。同时,固定化后的微生物细胞具有较好的重复利用性,连续反应3个批次后固定化细胞仍保留了80.0%以上的活性,而游离细胞的相对活性则小于20%。在所研究的体系中,葡萄糖为该反应的最佳辅底物,其最适浓度为50.0 mmol/L;该反应体系中的最适缓冲液p H值、反应温度、底物浓度分别为5.5、30℃和3.0 mmol/L。在此条件下,反应的初速度、产率以及产物的e.e.值依次为1.77 m M/h,88.9%和99.0%以上。     

YEAST AND THE VICTORIAN BREWERS: INCIDENTS AND PERSONALITIES IN THE SEARCH FOR THE TRUE FERMENT*

Events in the elucidation of the role of yeast in fermentation up to the end of the nineteenth century are described. It is concluded that, contrary to the popular view, the more enlightenend brewers had deduced the true nature of yeast prior to the time of Pasteur and were in advance of the dogma of the chemical establishment as represented by Liebig. The activities of scientists of the Victorian period employed in the Burton-on-Trent breweries are placed within this context.     

DIACETYL—A REVIEW: PART I—ANALYTICAL AND BIOCHEMICAL CONSIDERATIONS: PART II—BREWING EXPERIENCE

Diacetyl and 2,3-pentanedione are normal products of yeast metabolism and are formed in every brewery fermentation. The desired level in the final beer depends on the particular flavour aimed for but, in all types of beer, flavour defects are caused by excessive concentrations of diacetyl and many brewers might be happy to have no diacetyl in the beer. Recent improvements in analytical techniques show that many of the problems associated with diacetyl are due to the occurrence of compounds which can give rise to diacetyl in the finished beer. These compounds include the so-called “precursor,” acetolactic acid, but possibly other compounds such as the bisulphite addition compound of diacetyl are also involved. Study of the factors affecting diacetyl formation and removal by yeast shows how the concentration of diacetyl in beer can be controlled, and the processes at present used to regulate the diacetyl concentration in beer are described. The yeast strain used, the condition of the pitching yeast, the wort composition, the detailed management of the fermentation and the treatment of the beer during packaging and storage can all affect the diacetyl content of the beer.     

Relationships of Sensory Bitterness in Lager Beers to Iso‐α‐Acid Contents

Iso‐α‐acids and their chemically modified variants play a large role in evoking the bitter sensory attributes of lager character, but individual consumers may vary in their perception of bitterness. Sixteen lagers were scored in rank‐rating for bitterness by 14 trained assessors and the concentrations of the six bitter components in these beers were determined by high performance liquid chromatography. Relationships between bitterness intensity and the bitter components were modelled well using partial least square regression with a correlation value of 0.92. When 8 assessors carried out time‐intensity scoring of bitterness, profiles for single products were very different. However, single assessor profiles for multiple products showed qualitative similarities but quantitative differences. That individual assessors perceived bitter characters differently in relation to time has implications for new product development.     

METABOLIC QUOTIENTS OF RESPIRATION-DEFICIENT MUTANTS OF BREWER'S YEAST AND THE APPEARANCE OF A NEGATIVE PASTEUR EFFECT

Four industrial strains of bottom brewer's yeast and a group of their spontaneous respiration deficient (RD) mutants were tested for rates of metabolism of glucose and maltose under aerobic and anaerobic conditions. Q_o2 of all the RD mutants tested (26 isolates) ranged from 1·9 to 6·8 μl/mg yeast dry weight on glucose and was lowered to about one-half on maltose although the original strains had the same Q_o2 values on both sugars. No isolate showed any increase in glucose fermentation in aerobic conditions as compared with the original strains and the decrease of Pasteur effect found in certain isolates was always accompanied by a strong decrease of glycolysis in both aerobic and anaerobic conditions. Two mutants showed a strongly negative Pasteur effect, for their fermentation rates were higher in aerobic conditions than in anaerobic ones. Two other mutants showed a strong negative Pasteur effect only on maltose. The ratio of Q^N_2CO2 on glucose in most mutants was significantly lower than in their parental strains.     

SCREENING YEAST STRAINS FOR THEIR ABILITY TO PRODUCE PHENOLIC OFF-FLAVOURS: A SIMPLE METHOD FOR DETERMINING PHENOLS IN WORT AND BEER

Amounts of several phenolic compounds in wort and beer were measured by gas-chromatography following extraction with a small volume of chloroform. The procedure described is not specific for phenols and the limit of sensitivity is 0·05–0·1 μg ‘phenol’/ml. However, the precision is reasonable and this relatively rapid method has proved useful for screening yeast strains for their ability to produce phenolic off-flavours. Phenolic-tasting beers contained significant amounts of 4-vinyl guaiacol; whilst this compound may not be the sole cause of the undesirable flavour, its production is symptomatic of the unsuitability of a yeast strain for brewing.     

ANALYSIS OF HOPS AND HOP PRODUCTS FOR α-ACIDS OR ISO-α-ACIDS

Two methods based on the resolution of mixtures of hop compounds by chromatography on Sephadex columns have been adopted by E.B.C. and A.S.B.C. as ‘International Methods’.     

啤酒酵母中β-(1→3)-D葡聚糖的研究

叙述了啤酒酵母中碱不溶性β-(1→3)-D葡聚糖的结构特点和生物活性,对β-(→3)-D葡聚糖的提取工艺以及衍生化方法进行比较,并论述了β-(1→3)-D葡聚糖在水产养殖中的应用前景。     

FUNDAMENTALS OF IMMOBILISED YEAST CELLS FOR CONTINUOUS BEER FERMENTATION: A REVIEW

Recent fundamental research conducted on immobilised cells with a focus on continuous primary beer fermentation is presented in this review. The knowledge of whole-cell immobilisation, continuous fermentation, yeast biochemistry associated with beer flavour production, and bioreactor engineering design is required to apply immobilised yeast cells for industrial scale beer production. Understanding how immobilisation and continuous bioreactor operation affect yeast cell metabolism and viability will provide the groundwork for optimising beer quality. The latest studies on immobilised cell carriers, viability, vitality, mass transfer characteristics and bioreactor design indicate that an industrial scale immobilised cell system for primary beer fermentation may soon be a reality in the modern brewery .     

The α-aminoadipate pathway for lysine biosynthesis is widely distributed among Thermus strains

We previously reported that lysine is synthesized through the α-aminoadipate pathway in Thermus thermophilus HB27 (T. Kosuge and T. Hoshino, FEMS Microbiol. Lett., 169, 361–367, 1998), which was the first report demonstrating the synthesis of lysine through the α-aminoadipate pathway in Bacteria. LYS20 and LYS4, which respectively encode homocitrate synthase and homoaconitate hydratase have already been identified as the lysine biosynthetic genes in T. thermophilus HB27. In the present work, we examined eight other Thermus strains for the existence of genes belonging to the α-aminoadipate pathway. BamHI- or BglII-digested total DNAs from the eight strains were analyzed by Southern hybridization using LYS20 or LYS4 as a DNA probe. DNA fragments that hybridized with one or both of the genes were detected in seven of the Thermus strains but not in T. ruber. The sizes of the fragments that hybridized with the LYS20 and LYS4 probes were the same among T. thermophilus HB27, T. thermophilus HB8, “T. caldophilus” GK24, and four “T. flavus” strains. For example, a similar 4.3-kb fragment was detected in each of the above seven strains. In this fragment, four open reading frames were found downstream of the LYS4 gene in T. thermophilus HB27. Gene disruption experiments revealed that three open reading frames are involved in lysine biosynthesis in T. thermophilus HB27. These results strongly suggest that the lysine biosynthetic gene cluster for the α-aminoadipate pathway is widely distributed in the genus Thermus.