改进酿酒酵母谷物发酵性能的诱变研究

酿酒酵母经诱变获得有效利用麦汁糖的优良性能。突变菌株经5d摇床培养使YNB基础培养基的麦芽三糖发酵度从32.4%提高到81.8%。薄层层析证明突变株麦芽三糖转运能力得到改善。模拟工业发酵实验证明突变菌株的麦汁糖发酵性能大大改善,而原有发酵优良性状未受到影响。说明该突变株益于工业啤酒生产和低热啤酒的生产。     

低浓度淡爽型啤酒的酿造

以改善低浓度淡爽型啤酒品质为目的 ,提出了一种新颖的低浓度淡爽型啤酒的酿造方法 .采用二次煮出二段式糖化法 ,用 70 %麦芽和 3 0 %大米的原料配比 ,提高麦芽汁中糖与非糖的比值 ,并在糖化过程中添加啤酒酵母提取物作啤酒发酵的补充氮源 .所酿造的啤酒口味纯正 ,泡沫洁白细腻 ,持久挂杯 .     

Attachment of MAL32-encoded maltase on the outside of yeast cells improves maltotriose utilization

The fermentation of maltotriose, the second most abundant fermentable sugar in wort, is often incomplete during high-gravity brewing. Poor maltotriose consumption is due to environmental stress conditions during high-gravity fermentation and especially to a low uptake of this sugar by some industrial strains. In this study we investigated whether the use of strains with an alpha-glucosidase attached to the outside of the cell might be a possible way to reduce residual maltotriose. To this end, the N-terminal leader sequence of Kre1 and the carboxy-terminal anchoring domain of either Cwp2 or Flo1 were used to target maltase encoded by MAL32 to the cell surface. We showed that Mal32 displayed on the cell surface of Saccharomyces cerevisiae laboratory strains was capable of hydrolysis of alpha-1,4-linkages, and that it increased the ability of a strain lacking a functional maltose permease to grow on maltotriose. Moreover, the enzyme was also expressed and found to be active in an industrial strain. These data show that expressing a suitable maltase on the cell surface might provide a means of modifying yeast for more complete maltotriose utilization in brewing and other fermentation applications.     

Carrier‐free,continuous primary beer fermentation

Developing a sustainable continuous fermentation reactor is one of the most ambitious tasks in brewing science, but it could bring great benefits regarding volumetric productivity to modern breweries. Immobilized cell technology is often applied to reach the large densities of yeast needed in a continuous fermentation process. However, the financial cost associated with the use of carriers for yeast immobilization is one of the major drawbacks in the technology. This work suggests that yeast flocculation could address biomass immobilization in a gas‐lift reactor for the continuous primary fermentation of beer. Nearly 25 g dry wt L^?1 of yeast was flocculated in the reactor before interruption of the fermentation. Stable sugar consumption and ethanol production (4.5% alcohol by volume) from an 11°P wort was evidenced. The key esters and higher alcohols measured in the young beer met the standards of a finished primary beer fermentation. Copyright © 2014 The Institute of Brewing & Distilling     

Sourdough derived strains of Saccharomyces cerevisiae and their potential for farmhouse ale brewing

The Finnish farmhouse ale sahti is unique in that it is fermented with baking, rather than brewing strains of Saccharomyces cerevisae. The custom of maintaining farmhouse yeast cultures is however no longer practiced in Finland, and much yeast derived diversity in sahti beers has presumably been lost as a consequence. Here, the brewing potential of a number of sourdough derived strains was tested with respect to a number of different fermentation traits. Seven strains originally isolated from Finnish or Italian sourdough cultures were used to ferment high gravity sahti wort (20°P), and fermentation performance together with production of volatile compounds were assessed and compared with a reference baking yeast. Strains differed in terms of fermentation rate, yield, yeast viability and beer flavour profile. All were maltotriose positive, but utilisation varied so that alcohol yield could be greater or lower than that of the reference strain, with values ranging from 6.6 to 7.9% (v/v). Production of aroma compounds was also variable so that it was possible to identify strains producing high levels of esters and those with lower production, which could be used to emphasise flavours originating from raw materials. All strains generated 4-vinyl guaiacol and so would be suitable for other beers where this is a part of the normal flavour profile. Results suggest that sourdough isolates of S. cerevisiae are suitable for sahti production, but could also be applied to other beer styles as a way to differentiate products. © 2020 The Authors. Journal of the Institute of Brewing published byJohn Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling     

响应面法优化修道院强烈艾尔啤酒酿造工艺的研究

以皮尔森麦芽、特种麦芽和酒花为原料,选用上面艾尔酵母BE-256经二次发酵,酿制高酒精度的修道院强烈艾尔啤酒。 以感 官评分为评价指标,通过单因素试验研究发酵温度、加糖量及酒花干投量对啤酒感官品质的影响,在此基础上以感官评分为响应值, 建立响应面模型优化啤酒酿造的工艺条件。 结果表明,修道院强烈艾尔啤酒的最佳酿造工艺条件为发酵温度20 ℃,加糖量3.04%,酒 花干投量0.42 g/L。 在此优化条件下,啤酒的感官评分为92.8分,外观呈琥珀色,泡沫丰富细腻,香气宜人,酒体醇厚,杀口力强。     

三种不同酵母对啤酒发酵及风味的影响

本实验采用Y1110、安琪、模式三种不同的啤酒酵母,在同种工艺条件下测定发酵过程中α-氨基氮(α-AN)、pH、双乙酰、高级醇等指标,并比较三种不同啤酒的风味物质含量。结果表明:Y1110增殖最快,α-AN和pH值下降最快,双乙酰还原较快,后酵结束双乙酰含量最低,啤酒样品含醇量较高,适于醇厚型啤酒酿造;安琪酵母增殖最慢,α-AN和pH值下降最慢,啤酒样品含酯量较高,适于淡爽型啤酒酿造;模式酵母酯类与醇类含量都很高,不适于实际生产。     

Maltotriose utilization in lager yeast strains: MTT1 encodes a maltotriose transporter

Maltotriose is the second most abundant fermentable sugar in wort and, due to incomplete fermentation, residual maltotriose in beer causes both quality and economic problems in the brewing industry. To identify genes that might improve utilization of maltotriose, we developed a library containing genomic DNA from four lager strains and a laboratory Saccharomyces cerevisiae strain and isolated transformants that could grow on YP/2% maltotriose in the presence of 3 mg/l of the respiratory inhibitor antimycin A. In this way we found a gene which shared 74% similarity with MPH2 and MPH3, 62% similarity with AGT1 and 91% similarity with MAL61 and MAL31, all encoding known maltose transporters. Moreover, the gene shared an even higher similarity (98%) with the uncharacterized Saccharomyces pastorianus mty1 gene (M. Salema-Oom, unpublished; NCBI Accession No. AJ491328). Therefore, we named the gene MTT1 (mty1-like transporter). We showed that the gene was present in four different lager strains but was absent from the laboratory strain CEN.PK113-7D. The ORF in the plasmid isolated from the library lacks 66 base pairs from the 3'-end of MTT1 but instead contains 54 bp of the vector. We named this ORF MTT1alt (NCBI Accession No. DQ010174). 14C-Maltose and repurified 14C-maltotriose were used to show that MTT1 and, especially, MTT1alt, encode maltose transporters for which the ratio between activities to maltotriose and maltose is higher than for most known maltose transporters. Introduction of MTT1 or MTT1alt into lager strain A15 raised maltotriose uptake by about 17% or 105%, respectively.     

锌离子在啤酒酿造中的作用与控制

啤酒中锌离子来源于麦芽、大米、酿造用水、酒花。Zn^2 在啤酒酿造过程中可起到催化荆作用，与氨基酸结合形成Zn-氨基酸螯合物。在啤酒酿造过程中，可激活酶提高酶的作用；促进糖化、发酵；促进蛋白质合成及其稳定性；缓解某金属离子的毒性作用，促进挥发物质的产生和双乙酰的还原，缩短发酵时间，提高啤酒质量；但含量过量会使啤酒非生物稳定性降低，影响啤酒质量。通过对糖化过程和发酵过程的控制，可降低醪液pH值。加入少量小麦芽，加入适量ZnCl2，ZnSO4及酵母营养盐等，可实现对Zn^2 的有效控制，达到最佳酿造浓度。     

The Horace Brown Medal Lecture: Forty Years of Brewing Research

Horace Brown spent fifty years conducting brewing research in Burton‐on‐Trent, Dublin and London. His contributions were remarkable and his focus was to solve practical brewing problems by employing and developing fundamental scientific principles. He studied all aspects of the brewing process including raw materials, wort preparation, fermentation, yeast and beer stability. As a number of previous presenters of the Horace Brown Lecture have discussed Brown's achievements in detail, the focus of this paper is a review of the brewing research that has been conducted by the author and his colleagues during the past forty years. Similar to Horace Brown, fundamental research has been employed to solve brewing problems. Research studies that are discussed in this review paper include reasons for premature flocculation of ale strains resulting in wort underattenuation including mechanisms of co‐flocculation and pure strain flocculation, storage procedures for yeast cultures prior to propagation, studies on the genetic manipulation of brewer's yeast strains with an emphasis on the FLO1 gene, spheroplast fusion and the respiratory deficient (petite) mutation, the uptake and metabolism of wort sugars and amino acids, the influence of wort density on fermentation characteristics and beer flavour and stability, and finally, the contribution that high gravity brewing has on brewing capacity, fermentation efficiency and beer quality and stability.     

啤酒生产中酵母自溶及其预防措施

啤酒酵母自溶,影响啤酒质量。酵母自溶的原因有：①啤酒酵母品种和不正当地使用酵母;②糖化发酵不当;③过滤不好;④环境卫生不好。预防措施及解决办法有：①选用健壮、性能优良的茵种;②加强管理,调整糖化工艺;③搞好发酵环境卫生。     

Beer brewing using a fusant between a sake yeast and a brewer's yeast

Beer brewing using a fusant between a sake yeast (a lysine auxotrophic mutant of sake yeast K-14) and a brewer's yeast (a respiratory-deficient mutant of the top fermentation yeast NCYC1333) was performed to take advantage of the beneficial characteristics of sake yeasts, i.e., the high productivity of esters, high tolerance to ethanol, and high osmotolerance. The fusant (F-32) obtained was different from the parental yeasts regarding, for example, the assimilation of carbon sources and tolerance to ethanol. A brewing trial with the fusant was carried out using a 100-l pilot-scale plant. The fusant fermented wort more rapidly than the parental brewer's yeast. However, the sedimentation capacity of the fusant was relatively low. The beer brewed using the fusant contained more ethanol and esters compared to that brewed using the parental brewer's yeast. The fusant also obtained osmotolerance in the fermentation of maltose and fermented high-gravity wort well.     

啤酒酵母乙醛代谢关键酶相关性研究

乙醛是啤酒中的主要风味物质,其代谢主要来自酵母细胞。酵母中乙醇脱氢酶及乙醛脱氢酶是乙醛代谢的关键酶,对乙醛变化起着重要作用。跟踪啤酒酵母发酵过程中相对酶活力及乙醛变化,发现两种乙醇脱氢酶和乙醛脱氢酶的相对酶活力与发酵过程乙醛含量变化具有一定相关性。同时对低产乙醛啤酒酿酒酵母kb2-4与出发菌株啤酒酵母kb进行发酵试验,跟踪检测相对酶活力及乙醛含量,其乙醇脱氢酶Ⅰ和乙醇脱氢酶Ⅱ及乙醛脱氢酶相对酶活力均高于出发菌株,平均增幅分别为15.5%,11.6%和5%。3种酶活性的变化协同作用可以使乙醛含量降幅最大为33.8%。     

酿造水中的锌离子对啤酒酿造的影响

啤酒中的锌离子来源于麦芽、大米、酿造用水、酒花。实验表明在啤酒酿造过程中,锌离子可激活酶的活性、提高酶的催化作用,促进糖化、发酵;提高麦汁中糖、氨基酸的含量;促进双乙酰的还原,降低双乙酰的含量;激活乙醇脱氢酶,降低乙醛,提高酵母活力,降低酵母死亡率;提高发酵度,缩短发酵时间。     

高级醇对啤酒风味的影响及其在啤酒生产中的控制措施

高级醇是构成啤酒酒体的重要物质 ,是啤酒酿造过程中不可避免的副产物。高级醇赋予啤酒醇厚感、泡沫细腻 ,使啤酒丰满 ,但含量太高会破坏啤酒酒体及风味。影响和控制啤酒酿造过程中高级醇含量的因素有啤酒酵母、麦芽质量、麦汁成分和发酵工艺 (如发酵温度、发酵方法、发酵度 )等。     

混合微生物发酵生产啤酒醋及饮料开发

以大麦芽为主要原料,进行啤酒醋发酵,调配制备麦香啤酒醋饮料。优化后的啤酒醋发酵工艺条件为：麦芽糖化后接种啤酒酵母进行酒精发酵,24 h后接种生香酵母,继续发酵4 d,啤酒酵母和生香酵母的添加比例为4∶1;醋酸发酵阶段醋酸菌接种量为10%,30 ℃条件下发酵,发酵结束时啤酒醋酸度为（3.21±0.09） g/100 mL,乙醇含量＜0.05%vol。开发麦香啤酒醋饮料,最佳配方为啤酒醋12%,麦芽汁70%,白砂糖3%,橙色素0.04‰,最终产品的酸度为（0.38±0.07） g/100 mL,糖度为（84.32±0.05） g/L,酸糖比为1∶22。感官评定表明,该方法制备的麦香啤酒醋饮料色泽金黄、澄清透明、酸甜爽口,醋香和麦芽香气浓郁。     

Optimisation of High Gravity and Diet Beer Production in a German Brewery by Flow Cytometry

Increasing the quantity of beer production without diminishing the quality of the product is a key concern of the beer producing industry. Modifications to the brewery's equipment and settings are the most commonly used methods to improve the brewing process, while the supreme importance of the physiological state of the beer producing organisms, the yeast cells, for the productivity of the brewing process is often poorly recognised. The work described here was designed to optimise two processes: the inoculation regime used to produce high gravity bottom-fermenting beer, and the production of high quality diet beer. To achieve these aims, flow cytometry was used to follow changes in the distribution of DNA, neutral lipid and 3β-hydroxsterol contents in Saccharomyces carlsbergensis strains during inoculation, fermentation and storage. This allowed potential time-saving alterations in the process to be identified. Double staining techniques proved that vigorous fermentative activity and long-term survival capacity during main and secondary fermentation requires intense multiplication of the yeast cells during inoculation. The production of high gravity beer was then enhanced by altering the schedule of the wort additions, and thus increasing the yeast's activities related to multiplication. To produce diet beer, oligosaccharides that remain after the standard brewing process are degraded by adding small amounts of wort, usually during secondary fermentation. However, during this period of fermentation the physiological activity of the yeast cells is hampered by low carbon and high ethanol concentrations. Adding small batches of wort at carefully defined time points and in optimised amounts, even during the main fermentation, improves the physiological state of the yeast cells and rapidly decreases the carbon concentration within the fermentation tank. Both of these factors help to promote quick fermentation to a high quality diet beer. Thus, the flow cytometric investigations provided a reliable basis for identifying effective means of improving the process regime for brewing both of these products.     

啤酒发酵过程中酸类物质的变化与酵母活力关系的研究

研究了发酵过程中部分有机酸、脂肪酸含量的变化趋势与酵母活力的关系，以及使用不同代数的酵母对发酵液有机酸和脂肪酸含量、组成的影响；并且对比了不同使用代数酵母泥离心液和发酵液相应有机酸、脂肪酸含量的差值，以及差值与酵母活力的关系。     

浅论镁离子对啤酒发酵的影响

讨论了麦汁中镁离子含量对啤酒酵母发酵的影响。镁离子是酵母代谢过程中许多酶的重要辅助因子,它的浓度直接影响啤酒的发酵。提高镁离子的含量,均会提高初始发酵率,提高酵母活性,促进酵母对麦芽三糖及麦芽糖的吸收,增加酒精生成率和生成量,从而提高啤酒发酵度。     

THE EFFECTS OF GLUCOSE ADJUNCT IN HIGH GRAVITY FERMENTATION BY SACCHAROMYCES CEREVISIAE 2036

Laboratory fermentations of 16°Plato glucose adjunct worts by Saccharomyces cerevisiae 2036 demonstrated the absence of “glucose repression” of maltose and maltotriose uptake. However, when compared to worts in which maltose syrup was employed as an adjunct, residual glucose was present at the end of fermentation, maltose and maltotriose uptake rates were enhanced, fructose uptake was blocked and the sequence of sugar uptake was changed. These findings partially explain residual glucose and fructose that sporadically appear in commercial beers. Further research suggests that the physiological quality of the yeast is of prime importance in carbohydrate metabolism, and that critical concentrations of glucose vary with different physiological conditions for this brewing strain in 16°P wort .