激光诱变选育低双乙酰啤酒酵母

双乙酰是啤酒发酵过程中产生的一种重要的发酵副产物,也是衡量啤酒是否成熟的尺度。利用激光诱变选育双乙酰值低的啤酒酵母,得到一株优良菌株。用该菌株酿造的啤酒,其双乙酰值为0.1253mg/L。      

低双乙酰啤酒酵母的选育

将APV啤酒酵母菌株在不同的双乙酰含量梯度培养基内培养，挑出抗双乙酰的变异菌株，经菌种分离、筛选、发酵及双乙酰驯养等步骤选育出一株7^#菌，11℃低温发酵，双乙酰峰值0．36mg／L，成品啤酒双乙酰含量0．06mg／L，真正发酵度66．1％。     

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。      

低双乙酰啤酒酵母激光诱变条件的研究

以激光为诱变剂照射啤酒酵母，得到1株双乙酰值低的啤酒酵母。用该菌株酿造的啤酒，发酵液中双乙酰含量为0．0836mg／L，比用原菌株酿造的啤酒下降了53％。激光诱变的条件为：波长623．8nm(红色)、功率6mW的He—Ne激光，用滤纸片法照射啤酒酵母，照射时间为5min。     

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。     

低产杂醇油啤酒酵母菌株的选育

以啤酒酵母SC-4为出发菌株,经UV诱变育种后获得一株亮氨酸缺陷型菌株MS-11。与出发菌株相比,该菌株杂醇油生成量下降了25．47％,其他发酵性能基本保持不变。MS-11在较高主发酵温度(14℃)下的啤酒发酵实验表明,其杂醇油生成量为65．35mg／L,与出发菌株在9oC发酵条件下的杂醇油生成量63．24mg／L基本相当,而双乙酰的生成量则下降了15．13％。     

对富铁、富锌啤酒酵母代谢副产物的研究

于13.4°P 麦汁中分别添加不同含量的 FeSO_4·7H_2O 和 ZnSO_4·7H_2O,接入富铁、富锌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、pH、外观糖度、双乙酰、高级醇变化、后酵结束的各项理化指标:发现添加 FeSO_4·7H_2O 离子浓度为135.16ppm 的麦汁经富铁酵母发酵14天后,双乙酰含量为0.067mg/L,高级醇含量为56.2mg/L,酒精度为4.615,真正发酵度达67.6%;添加 ZnSO_4·7H_2O 离子浓度为166.38ppb 的麦汁经富锌酵母14天后,双乙酰含量为0.049mg/L,高级醇含量为59.1 mg/L,酒精度为4.670,真正发酵度为67.3%,啤酒风味基本不变,缩短了发酵周期,提高了产品质量。     

西藏灵菇中高产双乙酰乳酸菌分离筛选及产香特性研究

该研究旨在通过筛选具有较高双乙酰生产能力的乳酸菌改善酸奶香气品质。从西藏灵菇中分离筛选具有较高双乙酰生产能力的乳酸菌菌株并结合形态学特征和16S r DNA基因序列分析技术鉴定,利用气相色谱-质谱联用技术(GC-MS)对比分析筛选菌株发酵酸奶和空白酸奶香气品质差异。结果显示,菌株1-33具有较高的双乙酰生产能力,经鉴定其为植物乳杆菌(Lactobacillus plantarum)。菌株1-33制备的酸奶中双乙酰含量是空白酸奶的近4倍,乙偶姻的含量(38.92mg/L)是空白酸奶(25.46mg/L)的1.5倍左右,乙醛产量与空白发酵酸奶相当,说明该菌株具有优良的双乙酰生产能力,能改善酸奶的香气品质。     

激光-氯化锂复合诱变选育絮凝性适中的啤酒酵母

以絮凝性弱而其他发酵指标皆可的啤酒酵母菌Fs为出发菌株,以激光-氯化锂为复合诱变剂诱变啤酒酵母得到一株絮凝性适中的啤酒酵母,絮凝性为37.9%,比原菌株提高了1.61倍.用该菌株酿造的啤酒,发酵液中双乙酰含量为0.048mg/L,风味指标几乎不变.     

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。     

二次发酵降低葡萄酒中双乙酰的含量

该文通过在葡萄酒发酵后期添加蔗糖,促使酵母进行二次发酵降低双乙酰的含量。结果表明,二次发酵的最适加糖量为28 g/L,与常规发酵相比,二次发酵后双乙酰含量降低了29.87%;在此基础上,对酵母菌株WY1、WY1-1、WY1-2和WY1-12进行加糖二次发酵试验,测定出4株菌均可降低发酵液中双乙酰的含量,其中菌种WYI-12的发酵液中双乙酰的含量最低,为4.05 mg/L,比常规发酵降低了37.21%,改善了葡萄酒的感官质量。     

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。     

Fed‐batch fermentation with glucose syrup as an adjunct for high‐ethanol beer brewing

To produce a beer with a high ethanol content, preliminary research on fed‐batch fermentation profiles with glucose syrup as an adjunct during the primary fermentation period was conducted. The ethanol concentration of the beer was elevated by feeding a glucose syrup into the fermentors at a later stage of primary fermentation. Fermentation trials were carried out using a typical lager strain, SC‐9, with a pitching rate at 7.0 × 10⁶ cells/mL. An all‐malt wort (12.5°P) was employed and the primary fermentation temperature was 14 °C. Glucose syrup was supplemented when the concentration of residual reducing sugars was decreased to ~10 g/L. Results showed that the supplemented glucose was consumed rapidly and that the ethanol concentration in the final beer was raised to 67.9 g/L. Additional growth of yeast was observed after feeding accompanied by a low yield of ethanol (~0.46 g/g). Formation of diacetyl was enhanced by yeast growth and two additional peaks were obtained after feeding. The peak value of the diacetyl concentration was 1.90 mg/L. The fed‐batch fermentation resulted in a beer with an overproduction of higher alcohols and esters, indicating that brewing under these experimental conditions led to an unbalanced flavour profile. Results of optimization demonstrated that the optimal conditions were found to be 15°P for initial wort extract, 10 °C for fermentation temperature and 20 × 10⁶ cells/mL for yeast pitching rate, leading to total higher alcohols of 173.8 mg/L, total esters of 22.8 mg/L and an acetaldehyde concentration of 40.5 mg/L. A 12 day maturation and fermentation temperature of 8 °C was needed to reduce the acetaldehyde to 14.3 mg/L. Copyright © 2014 The Institute of Brewing & Distilling     

啤酒酵母融合株QSB—Ⅺ6中试研究

将新选育的酵母融合株ＳａｃｈａｒｏｍｙｃｅｓｃｅｒｅｖｉｓｉａｅＱＳＢ－Ⅺ６,同本厂生产中使用的酵母菌种ＳａｃｈａｒｏｍｙｃｅｓｃｅｒｅｖｉｓｉａｅＬＱ１６在２３０Ｌ发酵罐中做对比性发酵试验。试验酒的各项理化指标均已达到国家优质酒标准,其中双乙酰值在００８ｍｇ／ｌ以下,真正发酵度平均为７０２０％,酒体泡沫丰富,清亮透明。口味纯正,风味独特,为使该菌种能更好地应用于生产,奠定了基础。     

Analytical method validation and monitoring of diacetyl in liquors from Korean market

Diacetyl is a natural fermentation by-product and is an important flavor component of certain liquors. This paper aims to validate the high performance liquid chromatography (HPLC) method based on derivatization with 1,2-diaminobenzene for diacetyl quantification in liquor samples. A limit of quantitation of 0.039 mg/L was obtained. Coefficient regression (R²) of calibration curve for the HPLC–UV method exceeded 0.999, showing adequate linearity on the standard curve. Relative standard deviation values obtained from intraday and interday analysis for precision were 2.5 and 4.1%, respectively. Using the validated method, the contents of diacetyl in liquors (12 types, 389 samples) distributed throughout Korea were monitored. The average diacetyl content of all analyzed liquor samples ranged from trace amounts to 3.655 mg/L (microbrewery beer). The highest average diacetyl content was found in fruit wines (0.432 mg/L), followed by red wine (0.320 mg/L) and general distilled spirits (0.249 mg/L). In takju and yakju, no distinctive effect of sterilization on diacetyl content was found.     

传统藏式牦牛酸奶中优良乳酸菌筛选与产香性能分析

本研究旨在从我国传统发酵乳制品中筛选发酵性能和产香性能优良的乳酸菌。从四川阿坝藏族羌族自治州采集传统藏式牦牛酸奶样品并分离乳酸菌,经遗传稳定性、发酵时间及感官评价等初筛,并结合固相微萃取顶空气相色谱-质谱联用分析菌株产香性能进行复筛。结果显示,在26份样品中有5株发酵性能较优良的菌株,发酵时间为4.5~7.5 h,发酵终点酸度为74.0~78.4 °T,各菌株发酵乳感官评价良好。顶空气质结果发现菌株AB1504产生的乙醛(1.13 mg/L)、双乙酰(4.90 mg/L)等关键风味物质含量最高。研究结果表明从传统藏式牦牛酸奶中筛选到的嗜热链球菌AB1504发酵性能和产香性能优异,具有商业应用的潜力。     

两种不同啤酒酵母菌种发酵性能的比较

在本实验研究过程中采用两种不同酵母菌种APV、APK进行了对比试验，比较所生成的发酵副产物量的关系，特别是高级醇与双乙酰。在研究过程中发现，APV菌种发酵最后阶段的双乙酰值会出现反弹，而且含量高达0．20mg／L；APK菌种的双乙醚最后没有反弹现象，含量相比较而言，只有0．07mg／L。APV菌种发酵的成品啤酒高级醇含量为76．4mg／L，APK菌种的为53．4mg／L，且APK菌种的凝聚力较适中。     

Short communication: Combined antimicrobial activity of reuterin and diacetyl against foodborne pathogens

Reuterin (β-hydroxypropionialdehyde) is a broad-spectrum antimicrobial substance produced by some strains of Lactobacillus reuteri during anaerobic fermentation of glycerol. Some of these strains are able to survive and produce reuterin in cheese and yogurt when added as adjuncts to the starter. Similarly, in fermented dairy foods, other inhibitory compounds such as lactic acid and diacetyl are produced during fermentation. In this work, we studied the combined effect of reuterin and diacetyl under different pH conditions against Escherichia coli O157:H7, Salmonella Enteritidis, and Listeria monocytogenes. Results from agar spot assays showed that the antimicrobial activity of reuterin-producing strains against the gram-negative bacteria tested was enhanced as the concentration of diacetyl increased to 50 mg/kg, and was higher under acidic conditions (pH 5.0) for the 3 pathogenic strains. The combination of reuterin and diacetyl had an additive effect against L. monocytogenes only at diacetyl concentrations of 50 mg/kg and pH 5.0. In addition, growth kinetics studies showed that the combination of 1 activity unit (AU)/mL of reuterin with 100 mg/kg diacetyl increased the lag time of the 3 pathogens. In milk, synergistic antimicrobial activity was observed with the combination of 1 AU/mL reuterin and 50 or 100 mg/kg of diacetyl on the gram-negative strains tested, and with 1 AU/mL reuterin and 100 mg/kg of diacetyl on L. monocytogenes. The greatest inhibition of the 3 pathogens was achieved in acidified milk at pH 5.0 with reuterin (1 AU/mL) and diacetyl (100 mg/kg). Based on these results, the combination of reuterin and diacetyl in acidified dairy products could be a promising strategy to control food pathogens in these products.