啤酒发酵过程中影响蛋白酶A分泌的因素

纯生啤酒中残存的蛋白酶A严重影响泡沫稳定性,制约了纯生啤酒的质量提升。为了探索啤酒发酵过程中影响蛋白酶A分泌的因素,作者分别考察了菌种、酵母生理状态、酵母代数、麦汁浓度、发酵时间等对蛋白酶A分泌的影响。结果发现,蛋白酶A分泌量高的菌株,处于稳定期之后的酵母、较高的酵母代数、较高的原麦汁浓度和在发酵阶段末期都会导致发酵液中蛋白酶A活性偏高。建议在实际生产中,采用蛋白酶A分泌量少的菌种、调整酵母生理状态、使用小于3代的酵母、采用18°P以下的麦汁发酵和尽早结束发酵都会对降低蛋白酶A的分泌量起到积极作用。     

啤酒酿造中野生酵母的研究

啤酒酿造中野生酵母的研究吴文芳，张成刚，吕安国，李波（中国科学院沈阳应用生态所）金连武，蒋玲，孙喜春（辽宁铁岭市清河啤酒厂）所谓野生酵母，是指在啤酒酿造过程中出现的除培养酵母之外的所有酵母而言。威尔斯对野生酵母的定义是：任何未经严密选用与控制的酵母，...     

啤酒酿造过程中影响乙醛变化因素的研究

有针对性地提出了酿造过程中影响乙醛形成的生理内因和环境外因 ,并从主要的几个方面进行了研究。结果表明 ,酿造过程染菌、麦汁充氧控制不合理、二次发酵酒添加是造成成品啤酒中乙醛含量偏高的主要环境外因。在实际生产中 ,应加强CIP清洗 ,特别是种酵母罐的清洗及无菌空气过滤的控制工作 ,并合理控制麦汁的溶解氧含量 ,尽可能减少二次发酵酒的添加。     

高浓和低浓啤酒酿造中蛋白酶A对泡沫活性多肽的影响

啤酒产生丰富泡沫的能力受泡沫活性多肽水平的影响,含有疏水区域的特定多肽,如脂质转移蛋白(LTPl)是啤酒泡沫的重要组成成分。尽管高浓啤酒酿造在商业上是一项可行的技术,但同低浓酿造相比,其产品具有更低的泡沫稳定性。人们认为这主要是由于蛋白酶A对上述的疏水性多肽所起的降解作用造成的,本论文的研究目标是比较和确定在高浓(20°P)和低浓(12°P)麦汁发酵过程中疏水性多肽,尤其是泡沫-LTPl损失的数量,来评估蛋白酶A对这些多肽的影响,疏水性多肽和泡沫-LTPl,在高浓酿造中的损失更大,更进一步来说,已获得的结果表明蛋白酶A改变的是多肽的疏水性,而不是它们的分子量大小。大约有20％的疏水性多肽和57％的泡沫-LTPl表现出蛋白酶A抗性,这些疏水性多肽和泡沫-LTPl损失程度的差异直接影响到最终产品的泡沫稳定性。     

啤酒发酵过程中蛋白酶A的分泌及其与泡沫稳定性的关系

泡沫稳定性严重影响了纯生啤酒的质量。本文考察了蛋白酶A活性与泡沫稳定性的关系,结果显示二者呈明显负相关的关系。同时考察了影响蛋白酶A分泌的因素,结果发现,蛋白酶A分泌量高的菌株、处于稳定期之后的酵母、较高的酵母代数、较高的原麦汁浓度、发酵阶段末期、较低的酵母活力和较高的酵母死亡率都会导致发酵液中蛋白酶A活性偏高。因此在实际生产中,采用蛋白酶A分泌量少的菌种、调整酵母生理状态、使用小于3代的酵母、采用18°P以下麦汁发酵、尽早结束发酵、提高酵母活力、降低酵母死亡率都会对减少蛋白酶A的分泌量起到积极作用.     

啤酒废酵母酿造酱油的生产工艺

在啤酒的酿造过程中,将会产生大量的酵母副产物,这对于一个年产5万吨生产规模的中型企业来说,其废酵母浆可达750~1000吨.现在我国不少啤酒企业,都是将酵母浆直接投放到环境中去,这就造成了严重的环境污染。而实际上啤酒废酵母含48~55%的蛋白质、23~28%的碳水,化合物、6~8%的核糖核酸、2%的B族维生素、l%的谷胱甘肽,以及丰富的氨基酸     

酿酒酵母蛋白酶A的生化特性及生物学功能

酿酒酵母蛋白酶A是影响纯生啤酒泡沫稳定性的关键因素。由于纯生啤酒的广阔市场前景,对酵母蛋白酶A的研究越来越受到国内外研究者的关注。本文从蛋白酶A的结构、分泌、成熟、激活机制、催化机制、对啤酒泡沫的破坏作用以及酿造过程中蛋白酶A活性的变化等方面综述了蛋白酶A的研究进展,希望能为解决纯生啤酒泡沫稳定性问题提供理论参考。     

高浓发酵和酸洗对酿造酵母的影响

用磷酸类杀菌物质来洗涤接种酵母，以避免细菌污染是许多啤酒厂通用的方法，使用不正确的方法洗涤酵母将使其发酵特性下降并产生不良影响。酵母酸洗后接种于１２Ｐ麦汁，发酵特性没有明显下降；而接种于２０Ｐ麦汁，在发酵的第一个２４小时，酵母的活力就有所下降，但酵母的发酵特性没有变化。     

工业化酿造发酵过程中的非线性模型

本文主要研究了发酵过程中的非线性模型。由商业数据库得到的柏拉图糖度下降值随时间的变化趋势可以得出一个 S 形逻辑函数,方程中的四个发酵参数,即原终和真浓以及这个曲线的斜率和中点均由最小残差平方法得到。酵母的接种时间对发酵没有影响(P>0.05),起始温度能增加发酵速度(P<0.05),同时能降低达到发酵中点的时间(P<0.001),发酵起始温度正向影响啤酒真浓值(P<0.001)。本论文第一次说明了预测函数的预测区间结构,指出了发酵过程的预测偏差。本论文所用统计技术可以用来做发酵过程多方面的结果分析,例如能够预测当任何一个参数变化时酵母的增值倍数。这些技术可以用来通过统计四个发酵参数(如原浓、真浓、发酵曲线的斜率以及中点)的变化而了解发酵过程的变化(如温度、酵母和原浓),同时还可以用来评估不同的发酵过程,比如用一个新菌种发酵或用更高浓度的麦汁发酵。     

现代化啤酒酿造技术——固定化酵母工艺

<正> 自80年代发现适应酵母生存的高性能载体材料,酵母固定化技术在啤酒迮续发酵中的应用,再次引起人们的兴趣,然而,完整的啤酒连续发酵技术,仍然处于试验性阶段。至80年代后期,用于快速成熟和无醇啤酒生产的技术得到开发,并应用于工业上,使固定化酵母工艺再次成为啤酒工业的宠儿。最近,基伊埃集团Tuchenhagen研制出一种运用固定化酵母发酵罐所需的专业技术和设备,奉文将就固定化酵母连续发酵技术在一般和无醇啤酒生产中的应用进行探讨。     

Factors influencing proteinase A activity during the production of unpasteurised beer

Compared with pasteurised beer, a decline in foam retention during storage is an issue for unpasteurised beer. The major reason for this is that proteinase A is able to slowly breakdown foam promoting proteins in beer. Therefore, controlling the activity of proteinase A is key to solving this problem. In this study, foam quality in unpasteurised beer was studied systematically on a commercial scale considering factors including yeast activity, strain, generation number and storage time. Accordingly, yeast handling procedures to manage proteinase A activity were established: (1) yeast strain P with reduced proteinase A should be used in production; (2) storage time of recovered yeast should be no more than two days; (3) proteinase A activity in recycled yeast slurry should be less than 10×10^‐5 U/mL and (4) the number of yeast generations should be less than three. With the application of these measures, proteinase A activity was significantly decreased, and the corresponding foam quality was improved. © 2020 The Institute of Brewing & Distilling     

Purification of yeast proteinase A from fresh beer and its specificity on foam proteins

Yeast proteinase A from fresh beer was first purified with Sephadex G‐100 column chromatography and the active fractions reached to 5.3‐fold purification with 7% of yield. After purification with DEAE Sephadex A50, proteinase A activity increased to be 10.1 times of the initial with 1% of yield. When identifying the sample from chromatography by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE), only one protein band with 42 kDa was observed, this indicated that the enzyme was purified. The pattern of electrophoresis of hydrolysed beer by crude proteinase A did not show lipid transfer protein (LTP) on the gel. The result of SDS‐PAGE of interaction mixture of purified proteinase A and beer also indicated that LTP was decomposed.     

蛋白酶A对纯生啤酒泡持性的影响研究

该文研究蛋白酶A活力对纯生啤酒泡持性的影响,同时建立初始成品纯生啤酒及过滤前发酵液蛋白酶A活力限量值。结果表明,随着贮存时间的延长,纯生啤酒蛋白酶A活力和泡持值均呈下降趋势,最终残留蛋白酶A活力是影响其货架期泡持值的主要因素。纯生啤酒泡持值与蛋白酶A活力呈显著负相关（P＜0.01）。为了保证成品纯生啤酒在货架期内泡沫稳定性,发酵液出罐滤酒前的蛋白酶A活力应＜24×10-5 U/mL,相应成品纯生啤酒初始蛋白酶A活力应＜15×10-5 U/mL。该内控标准能为纯生啤酒生产企业控制泡持性提供指导。     

青岛啤酒酵母与高浓酵母筛选高浓酿造酵母融合亲株

以青岛啤酒酵母和高浓精酵母为供试菌株,筛选出生长良好的酵母,为选育具有青岛啤酒风味的高浓酵母做准备.比较了7株酵母不同糖类发酵、离子抗性、二氧化碳减重、发酵液风味品评等指标.结果表明:T1、T2和T3是传统的青岛啤酒发酵菌株,其发酵液口味符合青岛啤酒口味要求,且对Cu2+均不耐受;而G4和G6发酵减重试验和风味物质分析中的乙醛含量指标的评价均优于G5和G7菌株,且它们的发酵液的风味也接近青啤口味.因此,选择T1、T2、T3和G4、G6作100L酿造试验,进一步确定融合亲株.     

Construction of self-cloning industrial brewing yeast with high-glutathione and low-diacetyl production

Self‐cloning strains of industrial brewing yeast were constructed, in which one allele of α‐acetohydroxyacid synthase (AHAS) gene (ILV2) was disrupted by integrating Saccharomyces cerevisiae genes, γ‐glutamylcysteine synthetase gene (GSH1) and copper resistant gene (CUP1) into the locus of ILV2. The self‐cloning strains were selected for their resistance to CuSO₄ and identified by PCR amplification. The results of AHAS and glutathione (GSH) assay from fermentation with the self‐cloning strains in 500‐mL conical flask showed that AHAS activity decreased and GSH content increased compared with that of host yeasts. The results of pilot scale brewing in 5‐L fermentation tank also indicated that GSH content in beer fermented with self‐cloning strains T5‐3 and T31‐2 was 1.3 fold and 1.5 fold of that of host QY5 and QY31, respectively; and diacetyl content decreased to 64% and 58% of their hosts, respectively. The self‐cloning strains do not contain any heterologous DNA, they may be more acceptable to the public.     

固定化中性蛋白酶水解啤酒废酵母的研究

以离子交换树脂为载体,以成二醛为交联剂,固定化中性蛋白酶,再利用固定化的酶来水解啤酒废酵母。本文研究了不同水解时间和温度、不同的酶量对啤酒酵母水解的影响。通过研究发现,树脂styene—DVB D392对中性蛋白酶的固定化效果及固定化之后对酵母的水解效果比较好。     

The use of atmospheric and room temperature plasma mutagenesis to create a brewing yeast with reduced acetaldehyde production

High acetaldehyde levels in beer from yeast metabolism is a major concern for brewers in China. To obtain a strain with lower acetaldehyde production, this work reports a novel approach based on atmospheric and room temperature plasma mutagenesis and high‐throughput screening using 4‐methylpyrazole + disulphiram plating. A mutant LAL‐8a with lower acetaldehyde‐producing capability was obtained. The alcohol dehydrogenase activity decreased by 54% compared with the wild‐type M14 and the aldehyde dehydrogenase activity increased by 64% of the wild‐type strain. Through domestication and fermentation in EBC tubes, the mutant LAL‐8a was shown to produce 2.2 mg/L acetaldehyde, 88.2% less than the wild‐type strain M14. In addition, the ratio of higher esters to alcohols in beer fermented by the mutant LAL‐8a (0.28) was higher than M14 (0.16). The fermentation performance of LAL‐8a was similar to that of the wild‐type M14. This work suggests strain LAL‐8a a promising option for the brewing industry. © 2018 The Institute of Brewing & Distilling     

Reduced acetaldehyde production by genome shuffling of an industrial brewing yeast strain

High levels of acetaldehyde produced by yeast during fermentation can be of concern to product quality. A novel approach, based on genome shuffling, was applied to reduce the production of acetaldehyde by industrial brewing strain YS86. Four isolates with different impacts of acetaldehyde concentration were obtained from populations generated by ultraviolet irradiation and nitrosoguanidine mutagenesis. These yeast strains were then subjected to recursive pool‐wise protoplast fusion. A strain library that was likely to yield positive colonies was created by fusing the lethal protoplasts obtained from both UV irradiation and heat treatments. After two rounds of genome shuffling, a recombinant YSF2–9 strain produced less acetaldehyde than wild‐type strain YS86, by 64.5 and 66.2% in laboratory and pilot plant fermentations, respectively. The shuffled yeast strain YSF2–9 was genetically stable and may have a potential application in brewing industry for managing acetaldehyde in beer. Copyright © 2017 The Institute of Brewing & Distilling     

纯生啤酒泡沫稳定性的研究

纯生啤酒的泡持随着货架时间的延长会逐渐衰减 ,严重影响啤酒的外观质量。大量的文献资料证实 ,纯生啤酒泡持性的下降是由酒液中存在的蛋白酶A造成的。通过对成品酒泡持性的跟踪测定 ,重点讨论了发酵及啤酒过滤过程控制对泡持衰减趋势的影响。