啤酒酵母FLO8基因敲除对酵母发酵性能的改良

利用PCR介导基因中断技术,以pUG6为模板,设计含有与flo8基因两侧序列同源的长引物,构建带有卡那抗性基因（KanMX）中断盒,转化啤酒酵母G-03,获得一株转化菌G-03/f8。对转化菌G-03/f8和原菌G-03进行生理生化性能和摇瓶发酵性能比较。该菌株在实验室常规发酵中,生理性能、发酵性能基本上与出发菌株保持一致。当用酵母提前絮凝（PYF,premature yeast flocculation）值高的麦芽糖化后的麦汁接种发酵时,G-03/f8的絮凝性能与常规发酵下的絮凝性能基本一致,此时明显优于出发菌株G-03。G-03/f8的酒精度、发酵度等低温发酵指标与常规发酵相比有小幅度的下降,但明显高于此时G-03的各项发酵指标。相对于出发菌而言,G-03/f8对高PYF值的麦汁不敏感,能够保持较好的发酵性能,因此在高PYF值麦芽的利用上有良好的应用前景。      

酵母发酵性能试验机对啤酒酵母性能的测定

酵母发酵性能试验机是由管式发酵器（发酵管）和配套恒温循环仪两部分组成。本文通过酵母发酵性能试验机对啤酒酵母进行发酵性能的测定，提出其过程中的注意事项，对酵母的性能作出有效评估。     

高浓高温对啤酒酵母发酵性能的影响

以啤酒酵母S-6为实验菌株,研究了主发酵温度和原麦汁浓度对啤酒发酵的残糖、酒精度、风味物质和絮凝性等性能指标的影响。结果表明,原麦汁浓度一定时,主发酵温度对高级醇和乙酸酯的含量影响较大,主发酵温度由10 ℃提高至16 ℃时,高级醇含量提高了10%～20%,乙酸酯含量提高了8%～16%,但CO2累积质量损失、残糖、酒精度和絮凝性基本不受温度的影响;主发酵温度一定时,原麦汁浓度对酵母絮凝性影响较大,原麦汁浓度越高,酵母絮凝性越低,将高浓（18 °Bx）发酵液稀释50%至常浓（12 °Bx）,残糖、酒精度和高级醇的含量与常浓发酵液基本相同。该研究为选育高温高浓发酵低产高级醇同时强絮凝性酵母菌株提供了重要依据。     

德国啤酒酵母和国内啤酒酵母发酵性能的研究

酵母菌种是啤酒酿造的关键，不同的菌种可用来酿造不同类型的啤酒。本试验对德国酵母和国内酵母的发酵性能进行了对比研究，总结出了二者之间的差异。     

发酵工业菌种改良中的基因敲除策略及应用

基因敲除是一种近年来新发展起来的分子生物学技术,该技术可通过多种策略使机体特定基因缺失或失活,从而对生物遗传物质有目的地进行改造,在发酵工业微生物菌种改良中发挥着重要作用.在此对基因敲除技术的常用策略加以总结和论述,对其在工业菌种改良中的应用进行了简要介绍,同时展望了这项技术的发展趋势.     

几株酒精酵母的发酵性能比较

酵母菌种的好坏直接影响产品质量、设备利用率和产品产量，从而影响企业的经济效益。对1308菌株、1300菌株、安琪超酒酵母、奥特奇酵母产酒精的发酵、耐乙酸、耐乳酸、耐受酒精能力、耐受温度、耐渗透压、耐pH进行了发酵实验比较。结果表明，1308菌株、1300菌株、安琪超酒酵母、奥特奇酵母各有优缺点；菌株1308及安琪超酒酵母生产性能良好。     

空间诱变优良上面啤酒酵母发酵性能研究

以中德啤酒技术中心303上面啤酒酵母为出发菌株,将经过"神舟九号"宇宙飞船搭载进行航天诱变的303上面啤酒酵母菌进行分离得到若干菌株,对其中3株酵母的酵母凝聚性、发酵度、双乙酰还原能力、外观糖度变化、高级醇和酯类物质进行检测,从而选出1株优良的上面啤酒酵母菌。     

从发酵度指标差异入手探讨麦汁成分对酵母发酵性能的影响

本课题基于对数据的统计分析,从同样配方与工艺的酒发酵度出现悬殊差异现象入手,初步探讨了麦汁糖组分指标对酵母发酵性能的影响,并提出了减小真正发酵度指标波动的一项措施。     

压力对啤酒酵母生长及某些发酵性能的影响

以气体为加压介质,研究了不同压力下酵母细胞生长及压力对酵母发酵性能的影响.高压对酵母细胞的生长代谢有明显的影响:压力使酵母细胞的比生长速率和生物量降低,细胞的倍增时间延长;电子显微镜显示,压力使酵母菌细胞的形态发生明显的改变;压力也导致酵母的发酵速度、耗糖率、乙醇生成量、双乙酰的生成和还原速率下降.     

黄酒酵母HO基因的敲除及其对黄酒发酵的影响

为对黄酒酵母菌株的遗传学进行研究,对黄酒酵母利用基因敲除技术敲除HO基因,通过Mcclary产孢培养基于25?℃条件下培养5～7?d,得到a和α两种不同配型且配型不会发生转变的黄酒酵母单倍体菌株,通过群体杂交,成功获得了全敲除HO基因的二倍体酿酒酵母菌株黄酒酵母11-1-HOΔ,用于黄酒发酵实验。结果表明：通过基因工程手段敲除HO基因对黄酒发酵无显著影响,可用于工业生产中,且黄酒酵母11-1-HOΔ具有代表性,获得的单倍体是进一步研究黄酒酵母遗传基础和代谢机制的重要材料。     

Constructing an amylolytic brewing yeast Saccharomyces pastorianus suitable for accelerated brewing

An amylolytic brewing yeast Saccharomyces pastorianus, free of vector sequences and drug-resistance genes, was constructed by disrupting the alpha-acetolactate synthase gene and introducing the alpha-amylase gene as a selective marker. The resulting recombinant strain was able to utilize starch as the sole carbon source and its alpha-acetolactate synthase activity was lowered by 30%. Fermentation tests confirmed that the diacetyl concentration and the residual oligosaccharide were reduced by 70% and 25%, respectively, in fermented wort by the recombinant strain, while the brewing performance of the recombinant strain was retained.     

碱性和支链氨基酸对啤酒高浓酿造酵母发酵性能的影响

研究分别考察添加三种碱性氨基酸：精氨酸（Arg）、赖氨酸（Lys）、组氨酸（His）与支链氨基酸：缬氨酸（Val）、亮氨酸（Leu）、异亮氨酸（Ile）对24 °P高浓酿造过程中酵母发酵性能及啤酒风味的影响。结果表明,六种氨基酸的添加可促进酵母生长并提高活细胞率,显著提高麦汁发酵度及乙醇产量。其中,添加Val、Leu与Arg的高浓麦汁发酵性能较好,与对照组相比,发酵结束时总CO2失质量分别为92.4 g/L、92.7 g/L与91.2 g/L;发酵度极显著提高（P＜0.01）,分别为83.65%、82.95%和82.93%;乙醇产量极显著提高（P＜0.01）,分别为11.97%（V/V）、11.90%（V/V）和11.83%（V/V）。添加Arg可显著提高酵母总细胞数（1.64×108 cells/mL）（P＜0.05）,His、Val与Leu的添加对风味物质影响较大,可提高啤酒的醇酯比,其中,Val的添加可显著提高异丁醇含量（P＜0.05）,降低总酯含量;Leu的添加则可同时提高异戊醇及乙酸异戊酯的含量。     

酿酒酵母发酵性能的比较

以从民间自制黄酒酒药、酒厂的酒曲和窖泥中分离筛选得到的4株酵母(1#、2#、3#、4#为出发菌株,对其进行耐受酒精能力、耐受酸能力、发酵速率及发酵液中尿素含量、残糖、酒精度、总酸及氨基酸态氮等生理性能综合比较,其结果显示为4#酵母较其他3株发酵力强,耐受酒精浓度为14％vol,耐酸浓度为2.5％,产尿素含量为37.23mg/L,且其产尿素含量比2#酵母少56.7％,残糖浓度为12.57g/L,产酒精度达到16.1％vol,总酸含量为2.99g/L,氨基酸态氮含量为0.48g/L,具有进一步开发利用的价值.     

高浓酿造啤酒发酵周期的研究进展

高浓酿造是采用高浓麦汁发酵,之后进行稀释的啤酒酿造技术,该方法可显著提高生产效率,被广泛应用于啤酒生产中。虽然高浓酿造技术基本发展成熟,但仍存在发酵周期长以及后稀释造成风味不平衡的问题。由于高浓麦汁的特性,会导致发酵不彻底、发酵缓慢以至于延长发酵周期。该文分别从原料、发酵条件、菌株质量方面对高浓酿造啤酒发酵周期影响因素作了综合阐述,并对解决发酵周期长这一问题进行了讨论,提供可行思路。     

一株耐高浓度乙醇酒精酵母的筛选及发酵特性的研究

为获得乙醇耐受性较高的酵母菌,通过富集培养,从白酒窖池的酒糟中分离筛选出一株耐18%vol乙醇浓度的菌株A2。其最适生长温度为32℃,最适pH值为4.5。通过正交试验得出A2的最优发酵条件为葡萄糖浓度20%(w/v),温度34℃,pH值为4.5。发酵72h后,发酵液酒精浓度达到9.5%vol。     

Genetic and physiological characterization of yeast isolated from ripe fruit and analysis of fermentation and brewing potential

‘Wild’ and spontaneously fermented beers are growing in popularity in the craft beer industry. Most of these beers are fermented by the use of either pure cultures of unconventional yeast and bacteria or spontaneous fermentation using mixed local microflora. This study examined the potential of using pure strains of new isolates of wild yeast in the fermentation of a unique beer. The microbial communities from the fruit of pindo palm, loquat, hackberry and blackberry were collected in liquid culture, then plated for isolation. Ten isolates were selected for further analysis. Strains were identified by restriction fragment length polymorphism (RFLP) analysis and analysed for growth in a simple liquid media, fermentation in a complex media, alcohol tolerance and acid tolerance. Despite identification of some strains as the same species, they displayed a wide range of physiological properties. All strains were tolerant of pH values as low as 2.4, but none were tolerant of pH 1.9. Alcohol tolerance of different strains varied from 6 to 12%. Several strains had properties that suggest potential as primary fermenters, including the alcohol fermentation of a beer wort. Organoleptic properties of beers fermented with several of the strains demonstrated potential for commercial brewing. Copyright © 2014 The Institute of Brewing & Distilling     

High gravity primary continuous beer fermentation using flocculent yeast biomass

The current work assessed a new immobilized cell reactor system throughout a long‐term (54 days) continuous primary fermentation of lager‐type wort of high specific gravity. The experiment was performed in a 4 L airlift bioreactor and immobilization of biomass was attained solely by flocculation. Despite the constant liquid agitation and washout of biomass, up to 53 g dry wt/L of yeast remained immobilized in the system. Two types of beer were produced without interrupting the reactor, based on two types of wort: a Pilsener type with high specific gravity of 15.6 ± 0.3°P; and a dark lager wort with specific gravity of 14.4 ± 0.03°P. Even during the inlet of high gravity wort, the desired attenuation was achieved without the need for either recirculation or an auxiliary second stage bioreactor. The specific saccharide consumption rate was kept around 7.9 ± 0.4 g/L/h and ethanol productivity oscillated at 3.36 ± 0.2 g/L/h for nearly a month. During this period the volumetric productivity of the current bioreactor reached 1.6 L beer/L/day. The green beers produced from the Pilsener and dark lager worts met the standards of regular finished primary beer fermentation. The productivity of diacetyl through the entire experiment could be correlated to the free amino nitrogen consumption rate. Copyright © 2014 The Institute of Brewing & Distilling     

酵母多糖产生菌的筛选和发酵条件的研究

16种酵母菌通过初筛后，复筛时在培养基中加入磷酸二氢钾、碳源和氮源，筛选出高产多糖的酵母菌种——产脂酵母。确定此产脂酵母最佳发酵条件为：磷酸二氢钾0．6％，蔗糖6％，硝酸铵0．30％，玉米浆0．7％。     

不同氮源组成麦汁对酿酒酵母发酵性能的影响

研究了酿酒酵母Saccharomyces pastorianus(FBY0095)在六种不同氮源组成麦汁(20°P)中生物量、表观发酵度、乙醇浓度以及游离氨基氮(FAN)消耗量等指标的差异。结果表明,在20°P超高浓麦汁发酵中,麦汁氮源含量和组成对酵母发酵性能具有显著影响。随着麦汁中可同化氮量的提高,加速了酵母对糖的吸收利用,提高了酵母稳定期生物量和乙醇产量,发酵时间缩短了20%。当氮源匮乏(约为FAN=124.59 mg/L)时,补充适量大豆分离蛋白水解物(SPIH)于超高浓麦汁中,增加了麦汁氮源的多样性,使菌体增长量提高8.3%,乙醇产量提高4.43%,是酵母生长的有效氮源。