Stable disruption of ethanol production by deletion of the genes encoding alcohol dehydrogenase isozymes in Saccharomyces cerevisiae

We analyzed the effects of the deletions of genes encoding alcohol dehydrogenase (ADH) isozymes of Saccharomyces cerevisiae. The decrease in ethanol production by ADH1 deletion alone could be partially compensated by the upregulation of other isozyme genes, while the deletion of all known ADH isozyme genes stably disrupted ethanol production.     

High-temperature ethanol fermentation by immobilized coculture of Kluyveromyces marxianus and Saccharomyces cerevisiae

Suspended and immobilized cocultures of the thermotolerant yeast, Kluyveromyces marxianus DMKU 3-1042 and the mesophilic flocculent yeast, Saccharomyces cerevisiae M30 were studied for their abilities to improve production and stability of ethanol fermentation. Sugarcane juice and blackstrap molasses, at initial sugar concentrations of 220 g/L, were used as carbon sources. The results indicated that the coculture system could improve ethanol production from both sugarcane juice and blackstrap molasses when the operating temperature ranged between 33 °C and 45 °C. High temperature tolerances were achieved when the coculture was immobilized. The immobilized coculture was more effective in high-temperature ethanol fermentation than the suspended cultures. The coculture immobilized on thin-shell silk cocoon and fermented at 37 °C and 40 °C generated maximal ethanol concentrations of 81.4 and 77.3 g/L, respectively, which were 5.9-8.7% and 16.8-39.0% higher than those of the suspended cultures, respectively.     

3株酿酒酵母发酵过程中有机酸含量变化分析

通过高效液相色谱法对3株酿酒酵母(Saccharomyces cerevisiae)发酵过程中葡萄糖、甘油、乙醇及有机酸含量进行测定分析。结果表明:S.cerevisiae1946发酵体系中甘油和乙醇的含量显著高于S cerevisiae P1和S.cerevisiae 32788发酵体系(P0.05);3株酿酒酵母发酵液中7种有机酸存在明显的差异性和规律性;S.cerevisiae 1946发酵体系中有机酸的含量低于S.cerevisiae P1和S.cerevisiae 32788发酵体系,S cerevisiae的产酸水平与产乙醇能力呈负相关关系。     

Characterization and gene expression profiles of thermotolerant Saccharomyces cerevisiae isolates from Thai fruits

For industrial applications, fermentation of ethanol at high temperature offers advantages such as reduction in cooling costs, reduced risk of microbial contamination and higher efficiency of fermentation processes including saccharification and continuous ethanol stripping. Three thermotolerant Saccharomyces cerevisiae isolates (C3723, C3751 and C3867) from Thai fruits were capable of growing and producing 38 g/L ethanol up to 41°C. Based on genetic analyses, these isolates were prototrophic and homothallic, with dominant homothallic and thermotolerant phenotypes. After short-term (30 min) and long-term (12 h) exposure at 37°C, expression levels increased for the heat stress-response genes HSP26, SSA4, HSP82, and HSP104 encoding the heat shock proteins small HSP, HSP70, HSP90 and the HSP100 family, respectively. In isolates C3723 and C3867, expression was significantly higher than that in reference isolates W303 and TISTR5606 for TPS1 encoding trehalose-6-phosphate synthase, NTH1 encoding neutral trehalase and GSY1 encoding glycogen synthase. The results suggested that continuous high expression of heat stress-response genes was important for the long-term, heat stress tolerance of these thermotolerant isolates.     

发酵条件对一步法发酵菊芋生产乙醇的影响

使用产菊粉酶的酿酒酵母可实现一步法发酵菊芋生产乙醇.该试验以菊芋汁为发酵培养基,研究了起始糖浓度、无机盐、温度、种子培养基、接种量和氮源对高菊粉酶活酿酒酵母Y05-步发酵菊粉生成乙醇的影响.试验结果表明:(1)菊芋汁中最适起始总糖浓度为250g/L.菊芋汁含有的较多无机盐对乙醇发酵不但没有抑制作用,反而有促进作用;(2)最适乙醇发酵温度为37℃;(3)种子培养基中使用菊粉做为碳源有利于酿酒酵母Y05产生较多菊粉酶,进而促进其乙醇发酵;(4)最适乙醇发酵接种量为15％;(5)尽管菊芋汁是一个良好乙醇发酵培养基,但补加氮源仍是必要的.补加5.0g/L玉米浆可显著酿酒酵母Y05的提高总糖利用率、最终乙醇浓度和乙醇得率.     

在酿酒酵母中共表达sXYLA和XKS1及木糖发酵的初步研究

为改善重组酵母发酵木糖生产乙醇的能力,将定点突变改造后的Thermus thermophilus木糖异构酶基因sXYLA克隆到酵母表达载体pYX212并用于转化酸酒酵母Saccharomyces cerevisiae YPH499进行表达研究。酶活检测表明,改造后的木糖异构酶活性是未改造的1.91倍。在此基础上将改造后具有良好特性的木糖异构酶基因sXYLA和来自酸酒酵母的木酮糖激酶基因XKS1耦联,构楚得到重组表达质粒pYX-sXYLA- XKS1,在酿酒酵母YPH499中实现组成型共表达。结果表明,在84 h时重组菌发酵液酶活达到最高,木糖异构酶为0.624 U/mg蛋白,木酮糖激酶为0.688 U/mg蛋白。以葡萄糖和木糖为混合碳源初步进行半通氧发酵,代谢产物分析表明酸酒酵母重组菌木糖的消耗为4.75 g/L,乙醇的产量为0.839 g/L,分别比出发菌提高20.9%和14.8%,为酿酒酵母利用木糖发酵乙醇奠定基础。     

优良本土酿酒酵母的酿酒特性及产香能力初析

酿酒酵母产生的多种挥发性物质影响葡萄酒的香气风格,筛选本土酿酒酵母对改善葡萄酒同质化有积极作用.以13株不同来源的酿酒酵母和1株商业对照为试验菌株,对供试酵母的酒精耐受性、嗜杀性、产硫化氢能力及生长曲线进行了测定.随后在模拟汁中探究酵母的发酵特性,用固相微萃取-气相色谱/质谱(solid-phase microextr...     

Ethanol yield and volatile compound content in fermentation of agave must by Kluyveromyces marxianus UMPe-1 comparing with Saccharomyces cerevisiae baker's yeast used in tequila production

In tequila production, fermentation is an important step. Fermentation determines the ethanol productivity and organoleptic properties of the beverage. In this study, a yeast isolated from native residual agave must was identified as Kluyveromyces marxianus UMPe-1 by 26S rRNA sequencing. This yeast was compared with the baker's yeast Saccharomyces cerevisiae Pan1. Our findings demonstrate that the UMPe-1 yeast was able to support the sugar content of agave must and glucose up to 22% (w/v) and tolerated 10% (v/v) ethanol concentration in the medium with 50% cells survival. Pilot and industrial fermentation of agave must tests showed that the K. marxianus UMPe-1 yeast produced ethanol with yields of 94% and 96% with respect to fermentable sugar content (glucose and fructose, constituting 98%). The S. cerevisiae Pan1 baker's yeast, however, which is commonly used in some tequila factories, showed 76% and 70% yield. At the industrial level, UMPe-1 yeast shows a maximum velocity of fermentable sugar consumption of 2.27g·L(-1)·h(-1) and ethanol production of 1.38g·L(-1)·h(-1), providing 58.78g ethanol·L(-1) at 72h fermentation, which corresponds to 96% yield. In addition, the major and minor volatile compounds in the tequila beverage obtained from UMPe-1 yeast were increased. Importantly, 29 volatile compounds were identified, while the beverage obtained from Pan1-yeast contained fewer compounds and in lower concentrations. The results suggest that the K. marxianus UMPe-1 is a suitable yeast for agave must fermentation, showing high ethanol productivity and increased volatile compound content comparing with a S. cerevisiae baker's yeast used in tequila production.     

共培养时酿酒酵母对速生梭菌己酸代谢的影响及其机理

以1 株可以产己酸的速生梭菌（Clostridium celerecrescens）JSJ-1与1 株酿酒酵母（Saccharomyces cerevisiae）C-1为研究对象,在不同营养条件下,比较2 种微生物纯培养、共培养过程中生长代谢（菌落数、葡萄糖、乙醇、丁酸、己酸）的差异,分析S. cerevisiae对己酸菌己酸代谢的影响及其机理。结果发现：在厌氧条件下,34 ℃静置培养,S. cerevisiae C-1比C. celerecrescens JSJ-1更具有生长优势,会优先利用培养基中的葡萄糖。当培养基中唯一碳源为葡萄糖时,S. cerevisiae C-1会利用葡萄糖代谢生成乙醇,为C. celerecrescens JSJ-1合成己酸提供底物。当培养基中含0.5%葡萄糖和2%乙醇时,共培养相比C. celerecrescens JSJ-1单独培养,己酸的生成时间提前了4 d。葡萄糖对C. celerecrescens JSJ-1生成己酸有较强的抑制作用,共培养时S. cerevisiae C-1利用葡萄糖可缓解葡萄糖对己酸生成的抑制。在浓香型白酒发酵过程中,S. cerevisiae不仅可以为己酸菌合成己酸提供底物,而且可以缓解葡萄糖对己酸生成的抑制作用。     

以大豆糖蜜为原料高产乙醇酵母的筛选鉴定及其发酵特性研究

从大豆糖蜜中进行高产乙醇酵母的筛选和鉴定,并对其发酵特性进行研究。从大豆糖蜜中通过菌种的富集分离,TTC平板法初筛,耐乙醇能力及乙醇发酵能力的测定,筛选出一株乙醇产量达9.07%(V/V)的菌株P14。通过个体形态、菌落特征、生理生化及26S rDNA D1/D2区序列分析将菌株P14鉴定为酿酒酵母。研究了大豆糖蜜浓度及添加氮源和无机盐对酿酒酵母P14发酵生产乙醇的影响及酿酒酵母P14对大豆糖蜜中低聚糖的利用,结果表明大豆糖蜜浓度、添加氮源和无机盐对乙醇发酵影响显著,最佳的大豆糖蜜浓度为40%,添加氮源为1.2 g/L的蛋白胨;补加的无机盐为0.4 g/L MgSO4。在此培养基中发酵72 h后,糖蜜中90.10%的葡萄糖,91.23%的蔗糖,92.56%的棉籽糖和96.97%的水苏糖被酵母利用。因此大豆糖蜜中筛选出来的酿酒酵母P14具有较强的利用大豆糖蜜中的大豆低聚糖发酵产生乙醇的能力。     

工业酿酒酵母木糖代谢途径的建立及酒精发酵初步研究

以酵母AS2.1190为出发菌株,把含有木糖还原酶基因(XYL1)、木糖醇脱氢酶基因(XYL2)以及木酮糖激酶基因(XKS1)的质粒载体pYMIKP-xy127线性化后多拷贝整合进入其基因组,筛选得到基因工程菌株GZ4-127,并对此工程菌株进行葡萄糖、木糖共发酵试验.结果显示GZ4-127比出发菌株的菌体密度提高5%,木糖消耗提高2倍,酒精产率提高12%,说明工程菌已能够有效地利用木糖生产乙醇.     

乙醇对酿酒酵母Saccharomyces cerevisiae应激性的研究进展

在酿酒发酵工业中，虽然酿酒酵母Saccharomyces cerevisiae具有较高的乙醇耐受能力和乙醇产率，但也有一个乙醇的耐受能力极限。乙醇应激对酿酒酵母所产生的影响，包括乙醇应激对细胞质膜、HSP热激蛋白、抗氧化酶、海藻糖的影响以及高温高压等因素如何影响酿酒酵母的乙醇耐受力；以及当前研究中存在的问题。     

酿酒酵母的乙醇耐受性分子机理与全转录工程

研究酿酒酵母对乙醇耐受性的机理,对于发展乙醇生产有重要意义.酿酒酵母乙醇耐受性涉及到基因组水平上许多基因的复杂的相互作用,已知许多影响细胞膜的完整性和通透性、细胞壁结构、蛋白质构象,以及糖和氨基酸等的吸收等基因都与乙醇耐受性有关,与乙醇诱导相关的基因往往也与其他的环境因素如渗透压、热激、化学毒性、氧化压力等诱导的基因有关或重叠.因此,从基因转录动力学研究酿酒酵母乙醇耐受性并通过全转录工程构建乙醇耐受性工程菌己成为重要的研究热点.该文对近年来酿酒酵母乙醇耐受性分子机理以及全转录工程构建工程菌的研究作一综述,旨在为了解酵母乙醇耐受性机理和培育乙醇耐受性高产酵母菌株提供参考.     

绿色木霉和酿酒酵母降解稻草转化为乙醇的研究

以稻草作为碳源、污泥作为氮源,利用绿色木霉（Trichoderma viride）和酿酒酵母（Saccharomyces cerevisiae）降解稻草转化为乙醇,并对该过程中影响乙醇产量的因素进行优化。结果表明,当预处理稻草与预处理污泥质量比为1∶10（g∶g）、降解培养基初始pH值为7、绿色木霉和酿酒酵母的接种量分别为4%和3%、降解温度为37 ℃和降解时间为48 h时,可得到乙醇的含量为49.25 mg/L。     

冬果梨果酒酵母的筛选及鉴定

以冬果梨作为富集培养基,对野生酵母菌株进行收集、分离,得到980株;采用TTC平板法、耐乙醇性能测定以及产乙醇性能测定共三级筛选,最终筛得一株可耐受18%vol乙醇、乙醇产量达7.79%vol、发酵过程中糖利用率92.75%的菌株L-0301。通过对筛选所得菌株L-0301的形态特征观察、26S rDNA D1/D2区序列分析方法对菌株进行分类鉴定,结果显示,菌株L-0301为酵母亚科酵母属酿酒酵母菌（Saccharomyces cerevisiae）。     

海藻糖与酿酒酵母乙醇耐受性相关性的研究进展

为了得到具有高富锌能力的酵母菌,该研究以面包来源的酵母菌DLY28为出发菌株,重复驯化后筛选获得一株优良耐锌酵母,并通过单因素试验及响应面试验对其富锌培养基进行优化。结果表明,通过驯化筛选得到一株优良耐锌酵母S7,其富锌的最优培养基组成为蔗糖含量82 g/L、胰蛋白胨含量26 g/L、锌含量404 mg/L。在此最优条件下,富锌酵母S7的锌吸附量为18.79 mg/g,生物量（OD600 nm值）为1.49。该研究为富锌酵母的生产以及食品有机锌的开发应用提供理论依据。     

混合菌群降解稻草和污泥转化乙醇的研究

利用不同的混合菌群降解稻草和污泥转化乙醇。在初始pH值为7、降解温度为37 ℃的条件下,利用2 mL绿色木霉、1 mL短小芽孢杆菌和2 mL酿酒酵母降解1 g NaOH预处理稻草和10 g NaOH预处理污泥48 h,得到的乙醇产量为83.94 mg/L;在初始pH值为6、降解温度为37 ℃的条件下,利用3 mL绿色木霉、2 mL短小芽孢杆菌、3 mL酿酒酵母和2 mL树干毕赤酵母降解1 g NaOH预处理稻草和10 g NaOH预处理污泥48 h,得到的乙醇产量为49.29 mg/L。结果表明,绿色木霉、短小芽孢杆菌和酿酒酵母混合菌群能更有效地降解稻草和污泥转化乙醇。     

Cellular death of two non-Saccharomyces wine-related yeasts during mixed fermentations with Saccharomyces cerevisiae

The early death of two non-Saccharomyces wine strains (H. guilliermondii and H. uvarum) during mixed fermentations with S. cerevisiae was studied under enological growth conditions. Several microvinifications were performed in synthetic grape juice, either with single non-Saccharomyces or with mixed S. cerevisiae/non-Saccharomyces inocula. In all mixed cultures, non-Saccharomyces yeasts grew together with S. cerevisiae during the first 1-3 days (depending on the initial inoculum concentration) and then, suddenly, non-Saccharomyces cells began to die off, regardless of the ethanol concentrations present. Conversely, in both non-Saccharomyces single cultures the number of viable cells remained high (ranging 10(7)-10(8) CFU ml(-1)) even when cultures reached significant ethanol concentrations (up to 60-70 g l(-1)). Thus, at least for these yeast strains, it seems that ethanol is not the main death-inducing factor. Furthermore, mixed cultures performed with different S. cerevisiae/ H. guilliermondii inoculum ratios (3:1; 1:2; 1:10; 1:100) revealed that H. guilliermondii death increases for higher inoculum ratios. In order to investigate if the nature of the yeast-yeast interaction was related or not with a cell-cell contact-mediated mechanism, cell-free supernatants obtained from 3 and 6 day-old mixed cultures were inoculated with H. guilliermondii pure cultures. Under these conditions, cells still died and much higher death rates were found for the 6 days than for the 3 day-old supernatants. This strongly indicates that one or more toxic compounds produced by S. cerevisiae triggers the early death of the H. guilliermondii cells in mixed cultures with S. cerevisiae. Finally, although it has not been yet possible to identify the nature of the toxic compounds involved in this phenomenon we must emphasise that the S. cerevisiae strain used in the present work is killer sensitive with respect to the classical killer toxins, K1, K2 and K28, whereas the H. guilliermondii and H. uvarum strains are killer neutral.     

混合菌同步糖化共发酵造纸污泥产乙醇

对酿酒酵母（Saccharomyces cerevisiae）与重组大肠杆菌K011（Escherichia coli）混合菌同步糖化共发酵造纸污泥产乙醇进行了初步研究。在底物浓度为50g／L时,通过单因素实验和正交实验获得乙醇发酵的最佳条件：纤维素酶添加量25FPU／g底物,接种量为6％,酿酒酵母与重组大肠杆菌K011接种比例为1：1（细胞干重初始浓度分别为1．0g／L和0．3g／L左右）。发酵72h后,乙醇浓度为5．71g／L,产率达到0．114g乙醇／g污泥,达到理论值的42．5％。分别用酿酒酵母、K011单菌种发酵与双菌株组合发酵对比结果表明,混合菌发酵效果明显优于单菌种发酵。     

酒精与化妆品

简要介绍了酒精的生产方法、生产原料及分类,并从化妆品的角度介绍了酒精的应用情况,同时指出了酒精是化妆品的必要原料,只要正确使用,对人体无害。