双亲灭活的原生质体融合株啤酒酵母DR9-2的构建及其特性的研究

啤酒酵母菌株JW1-1发酵液中的双乙酰、总高级醇的含量较低,但发酵度较低,发酵液中的乙醛含量较高;啤酒酵母菌株NW7-45发酵度较高,发酵液中的乙醛含量较低,但发酵液中的双乙酰、总高级醇的含量较高.为了得到优良的菌株,以紫外线灭活的菌株JW1-1原生质体和热灭活的菌株NW7-45原生质体为亲本进行融合.经三角瓶发酵筛选,得到较优良的融合株DR9-2和DR9-24.其中融合株DR9-2以12°Bx麦芽汁为培养基,用500L的发酵罐在12℃下发酵,发酵11d发酵液的发酵度为69.5%,发酵液中的双乙酰、乙醛和总高级醇的含量分别为0.0124mg/L、7.70mg/L和61.88mg/L.从融合株DR9-2的主要发酵特性和啤酒口感品评的结果表明,该菌株在啤酒酿造工业中具有应用前景.     

啤酒酵母原生质体融合株GR8的中试研究

啤酒酵母原生质体融合株GR8的中试表明:融合株GR8凝絮性较强(本斯值为3.0);以12oBx麦芽汁为培养基,用500L发酵罐在12℃下发酵15d,发酵度为69.6%;发酵液中的双乙酰、乙醛、总高级醇、乙酸乙酯和乙酸异戊酯的含量分别为0.0390mg/L,3.26mg/L,84.7mg/L,11.14mg/L和0.99mg/L。     

高浓度双乙酰定向驯化获得优良啤酒酵母菌株的研究

在双乙酰浓度为1mg/mL的12°Bx麦芽汁中进行高浓度定向驯化,经涂布抗双乙酰固体选择培养基,从啤酒酿造生产菌株啤酒酵母(Saccharomycessp.)A中富集筛选分离得到一株双乙酰还原速度优于亲株A的新菌株TA4。以12°Bx麦芽汁为培养基用内装300mL麦芽汁的500mL三角瓶10℃下发酵,发酵8d后发酵液双乙酰含量比亲株降低了36.31%,发酵度提高4.5%,且该菌株的絮凝性、发酵速率等特性仍保持了亲株A的优良性状。     

高浓度双乙酰定向驯化获得优良啤酒酵母菌株的研究

在双乙酰浓度为1mg/mL的12°Bx麦芽汁中进行高浓度定向驯化,经涂布抗双乙酰固体选择培养基,从啤酒酿造生产菌株啤酒酵母(Saccharomycessp.)A中富集筛选分离得到一株双乙酰还原速度优于亲株A的新菌株TA4。以12°Bx麦芽汁为培养基用内装300mL麦芽汁的500mL三角瓶10℃下发酵,发酵8d后发酵液双乙酰含量比亲株降低了36.31%,发酵度提高4.5%,且该菌株的絮凝性、发酵速率等特性仍保持了亲株A的优良性状。      

啤酒酿造酵母原生质体融合株GR5的中试

报道了啤酒酵母原生质体融合株GR5的中试结果。融合株GR5的凝絮性较强 (本斯值为 2 .7) ,以12°Bx麦芽汁为培养基 ,用 5 0 0L发酵罐在 12℃下发酵 ,发酵至第 8d菌株GR5的发酵度为 6 9.2 % ,发酵液中的双乙酰含量为 0 .0 4 98mg/L、乙醛含量为 6 .34mg/L ,总高级醇含量为 74 .4mg/L ,该菌株生产的啤酒 ,口感柔和、协调 ,略带酯香味。融合株GR5的遗传性稳定 ,是一株具有工业应用前景的啤酒酿造酵母菌株     

啤酒酵母融合株GR8发酵特性的初步研究

研究了啤酒酵母原生质体融合株GR8的主要发酵特性。以12°Bx麦芽汁为三角瓶发酵培养基,在12℃下发酵8d,融合株GR8的发酵度为66.5%,凝絮性(本斯值)为3.0mL;发酵液中的双乙酰、乙醛、高级醇和乙酸乙酯等风味物质的含量分别为0.0583mg/L、9.66mg/L、91.20mg/L和22.8mg/L。从主要发酵特性的指标和口感品评以及遗传稳定性的结果表明,融合株GR8是一株具有工业应用前景的啤酒酿造酵母菌株。     

化学诱变选育低双乙酰啤酒酵母菌株的研究

通过EMS诱变,从啤酒酿造生产菌株啤酒酵母(Saccharomyces carlsbergensis)FB中筛选分离得到一株发酵液中双乙酰含量优于亲株的新菌株FB-E1。以120Bx麦芽汁为培养基,用内装300ml麦芽汁的500ml三角瓶于12℃下发酵,发酵8d后发酵液中双乙酰含量比亲株降低了42.7%。该菌株的其它发酵性能的测定结果表明其保持了亲株的优良性状,且遗传性状稳定。     

富硒麦汁对啤酒酵母代谢副产物的影响

于11°P富硒麦汁中接入啤酒酵母进行常规啤酒发酵,同时以空白麦芽汁发酵作为对照。在整个发酵过程中,跟踪检测酵母生长情况;并测定发酵14天后发酵液的pH、外观糖度、双乙酰、高级醇含量、后酵结束各理化指标,发现添加Na2SeO3含量为200mg／L的富硒麦芽制成的麦汁经酵母发酵14天后,双乙酰含量为0．07mg/L,高级醇含量为63．2mg／L,酒精度为5．95％,真正发酵度达74．5％,啤酒风味基本不变,缩短了发酵周期。     

用分子生物学手段控制啤酒中乙醛含量的研究

以F718,R719为引物,以质粒pFA6a—kanMX4为模板进行PCR扩增,采用基因转化法获得1株乙醇脱氢酶Ⅱ基因突变型工业酿酒酵母。驯养后的突变株啤酒生产小试表明,突变株乙醛含量为5．386mg／L,比原菌株乙醛含量7．932mg／L有所降低;发酵液发酵结束时,双乙酰含量为0．058mg／L,比原菌株双乙酰含量0．034mg／L有所升高;突变株发酵度为63％,比原菌株66％略有降低。     

麦芽酵母超前絮凝多糖对啤酒发酵液质量的影响

研究了麦芽PYF(酵母超前絮凝)多糖对啤酒发酵液质量的影响.测定了各实验组发酵液的发酵度、酒精含量、风味物质及有机酸含量,发现PYF多糖发酵可使最终发酵液中悬浮的酵母数量降低,发酵液的发酵度和酒精含量降低,双乙酰和乙醛含量升高,高级醇、酯类、二甲基硫和有机酸含量降低.     

100%大麦啤酒酿造过程中老化Strecker醛的研究

分别采用上面发酵工艺与下面发酵工艺进行100%大麦啤酒及100%麦芽啤酒的酿制,并对其麦汁的氨基酸含量、老化Strecker醛、自由基以及新鲜啤酒中老化Strecker醛的含量等进行了对比分析。研究发现,就麦汁而言,100%大麦麦汁中老化Strecker醛的含量都明显低于100%麦芽麦汁;同样的麦汁,上面发酵方式还原Strecker醛的能力明显优于下面发酵方式。就啤酒而言,经酵母还原后,新鲜啤酒中的老化Strecker醛含量较麦汁含量低,且100%大麦啤酒中老化Strecker醛的含量低于100%麦芽啤酒中的含量。100%麦芽麦汁的自由基含量是100%大麦麦汁的近3倍。这都预示着100%大麦啤酒的风味稳定性（新鲜度）明显好于100%麦芽啤酒。     

3-甲硫基丙醛对啤酒麦汁味的影响及其GC/MS定量检测方法的建立

对3-甲硫基丙醛在啤酒中的风味特点及对啤酒麦汁味的影响进行了研究,并对其在啤酒中的阈值进行了测定,研究结果表明啤酒中3-甲硫基丙醛具有类似于谷物蒸煮的味道,描述为麦皮味,与麦汁味相近,且其含量的高低与麦汁味成显著正相关,可以用3-甲硫基丙醛作为表征啤酒麦汁味的指标。经感官品评测定其在啤酒中的差别阈值为5.0μg/L。对样品进行蒸馏萃取结合GC-MS,建立了一种测定麦汁或啤酒中3-甲硫基丙醛的方法,采用外标法定量,该法的相关系数为0.9952,精密度为4.57%,回收率为83.71%～92.48%,检出限为0.015μg/L。应用这一方法检测了不同啤酒中3-甲硫基丙醛的含量,发现普通啤酒都在50μg/L以下,结合其差别阈值,得到啤酒中3-甲硫基丙醛的控制标准为不超过55μg/L。     

利用植物乳杆菌进行啤酒生物酸化的初步研究

从长期发酵的啤酒中分离到1株乳杆菌,经鉴定为植物乳杆菌。对该植物乳杆菌的发酵产酸特性进行了初步研究,并将其应用到麦汁糖化中,利用生物酸化技术对麦芽汁进行调酸,经实验室微型啤酒糖化发酵设备生产,可得到质量优良的绿色啤酒。     

反相高效液相色谱法(RP-HPLC)测定啤酒中有机酸

建立了一种利用反相高效液相色谱法同时测定啤酒中 9种有机酸的方法。应用反相C18色谱柱Nucleosil 10 0C18　 2 5 0× 4 0mm ,以 0 1mol/LKH2 PO4(用H3 PO4调 pH3 0 )为流动相 ,紫外检测 2 15nm ,将啤酒中草酸、丙酮酸、苹果酸、α 酮戊二酸、乳酸、乙酸、柠檬酸、富马酸、琥珀酸一次性分离。除柠檬酸外各种有机酸的回收率在 85 %以上 ,变异系数 <5 %。该方法简便、快速、准确。此外 ,对影响有机酸分离的因素进行了研究 ,并测定了国内市售的十几种啤酒中的有机酸含量     

Construction of a Single PEP4 Allele Deletion in Saccharomyces carlsbergensis and a Preliminary Evaluation of Its Brewing Performance

The secretion of proteinase A (encoded by PEP4) from brewer's yeast is detrimental to the foam stability of unpasteurized beer. The aim of this study was to construct mutants of the allopolyploid Saccharomyces carlsbergensis strain TT, which were partially or completely deficient in proteinase A activity. Allelic PEP4 genes were consecutively disrupted by using the Cre‐loxP recombination system combined with PCR‐mediated gene disruption. A single PEP4 deletion mutant TT‐M was successfully constructed. However, no viable mutant could be obtained when the second allelic PEP4 gene was deleted. The brewing performances of the parent strain and the modified strain were compared on a 100 L pilot fermenter scale. Proteinase A activity in fermented wort brewed with mutant strain TT‐M was significantly lower (p<0.05) than that of the parent strain TT, whereas no significant difference on either maltose or maltotriose assimilation (p>0.05) was found. The mutant TT‐M remained genetically stable, as shown by diagnostic PCR, after re‐streaking for 20 generations. The flavor and taste of the final fermented wort, brewed with the mutant strain TT‐M, was evaluated by the Tsingtao expert sensory panel, and found to be comparable to that of the parent strain and exhibited no distinct defects. The flavor component profiles of these two finished products were also comparable. The study demonstrated allelic genes in polyploid industrial yeasts could be efficiently and consecutively deleted by the retractive primer disruption strategy, and the mutant of Saccharomyces carlsbergensis partially deficient in proteinase A contributed to an improvement in foam stability.     

Mutagenizing brewing yeast strain for improving fermentation property of beer

A brewing yeast mutant with perfect sugar fermentation capacity was isolated by mutagenizing the Saccharomyces pastorianus transformant, which carries an integrated glucoamylase gene and has one copy of non-functional alpha-acetolactate synthase gene. The mutant was able to utilize maltotriose efficiently, and the maltotriose fermentability in YNB-2% maltotriose medium increased from 32.4% to 72.0% after 5 d in shaking culture. The wort fermentation test confirmed that the sugar fermentation property of the mutant was greatly improved, while its brewing performances were analogous to that of the wild-type strain and the characteristic trait of shortened beer maturation period was retained. Therefore, we believe that the brewing yeast mutant would benefit the beer industry and would be useful for low caloric beer production.     

西瓜啤酒制作工艺的研究

以西瓜汁和大麦芽为主要原料,采用啤酒独特的低温发酵工艺酿制西瓜啤酒。通过单因素试验和响应面分析试验,以感官评分为评价标准,结合理化指标研究了西瓜汁添加量、西瓜汁添加时间和原麦汁浓度对西瓜啤酒品质的影响。确定了西瓜啤酒的最佳发酵工艺条件为：西瓜汁添加量14.0%,在发酵的第4.5天添加西瓜汁,原麦汁浓度11.2°P。在此条件下,进行了400 L发酵罐的验证试验,获得了一种具有西瓜清爽口感和啤酒典型口感的西瓜果味啤酒。     

Effects of β‐Glucans and Environmental Factors on the Viscosities of Wort and Beer

This paper reports on the influence of molecular weight and concentration of barley β‐glucans on the rheological properties of wort and beer. Environmental conditions such as pH, maltose level in wort, ethanol content of beer, shearing and shearing temperature were also examined for their effects on wort and beer viscosities. In the range of 50–1000 mg/L, β‐glucans increased solution viscosity linearly with both molecular weights (MW) of 31, 137, 250, 327, and 443 kDa and concentration. The influence of MW on the intrinsic viscosity of β‐glucans followed the Mark‐Houwink relationship. Shearing wort and beer at approximately 13,000 s^?1for 35 s was found to increase the wort viscosity but reduce beer viscosity. Shearing wort at 20°C influenced β‐glucan viscosity more than shearing at 48°C and 76°C whereas the shearing temperature (0, 5 and 10°C) did not effect the viscosity of beer. At lower pHs, shearing was found to reduce the viscosity caused by β‐glucans in wort but had no effect in beer. Higher concentrations of maltose in wort and ethanol in beer also increased the viscosity of β‐glucan polymers. It was found that β‐glucans had higher intrinsic viscosities in beer than in wort (5°C), and lower critical overlap concentrations (C^*) in beer than in wort.     

啤酒发酵过程中甲醛代谢影响因素初步研究

甲醛是啤酒酵母的一种代谢产物,在啤酒发酵过程中,甲醛经历了积累和随后的还原两个阶段,通过引入甲醛峰值和谷值两个概念研究了啤酒生产过程中的一些影响甲醛积累和还原的控制,结果表明:(1)过低或过高的麦汁溶氧量均会增加甲醛的积累,在12～13mg/L的麦汁溶氧条件下甲醛积累最小。(2)低代酵母甲醛积累要少于高代酵母,并且低代酵母在后期甲醛的还原能力要高于高代酵母,酵母接种量在2×107个/ml时甲醛峰值最低。(3)相对较高的还原温度有利于甲醛还原。(4)麦汁浓度对甲醛的积累影响较大,高麦汁浓度条件下的甲醛峰值要比低麦汁浓度高出许多。(5)啤酒酿造后期延长还原时间并不能显著增加甲醛的还原量。     

啤酒酵母代谢副产物高级醇的影响因素研究进展

高级醇是啤酒酿造过程中产生的副产物的主要成分,是啤酒的主要香味和口味物质之一。高级醇的生成途径有降解代谢和合成代谢两种。适量的高级醇能赋予啤酒丰满的香味和口味,增加酒体的协调性,但过量存在也是啤酒异杂味的主要来源之一。高级醇含量超过100mg／L会使啤酒口味和受欢迎程度明显降低,啤酒中高级醇含量的标准值为：下面发酵啤酒60～90mg／L;上面发酵啤酒〈100mg／L。高级醇含量主要受酵母菌种、麦汁成分以及发酵工艺条件的影响．因此生产过程应控制好这几个因素。