应用PCR-DGGE技术分析酱香型白酒酒曲细菌多样性

利用PCR-DGGE技术研究酱香型白酒制曲过程中的细菌菌群结构及其消长规律,鉴定出不同样品的优势茵群。结果表明,酱香型大曲间的细茵组成存在明显差异,母曲与出仓曲的相似性系数仅为O．29。随着曲药的发酵,细菌多样性下降,优势茵群变化明显。其中,芽孢杆菌属（Bacillus）、明串珠菌属（Leuconostoc）、片球菌属（Pedio-cocuss）、魏斯氏菌属（Weissella）、棒状杆菌属（Corynebacterium）、高温放线茵属（Thermoactinomyces）和乳酸杆菌属（Lactobacillus）是母曲和翻仓曲的优势菌群,而出仓曲的主要类群为芽孢杆菌属（Bacillus）和乳酸杆菌属（Lactobacillus）。另外,还发现了在白酒曲药中不曾报道的种群和不能培养的细茵：Uncultured Propionibacteriaceae bacterium、Uncultured Weissellasp、Uncultured cyanobacterium,该技术克服了传统分离培养的缺点。     

PCR-DGGE指纹技术在食品微生物研究中的应用

分子生物学的发展带动了实验技术的进步，DGGE作为一种不依赖于培养的分子技术最近被用于食品微生物的研究。本文简要介绍了DGGE基本原理，及其在微生物鉴定，种群多样性研究和发酵过程动态监测中的应用，并对该技术自身存在的缺陷进行了评价。     

浓香型白酒酒醅真菌群落结构及其变化规律

利用PCR-DGGE技术研究了浓香型白酒酒醅真菌群落结构及其发酵过程中的变化规律,结果发现,所有酒醅样品均出现9～19条较清晰的条带,其中第2、5、8、9、14、17号条带在所有样品中均有检出,且优势度较高;所有样品生物多样性指数都在1.68～2.61之间。发酵前期,多样性指数呈先下降后上升的趋势,发酵至第21～49天基本保持平稳,第56天时,多样性指数到达最低值,第63天回升之后多样性指数持续下降,发酵至第84天时略有回升;不同发酵时期酒醅真菌DGGE图谱相似性指数较低,表明不同发酵时间酒醅样品真菌群落间差异较大。     

PCR-DGGE技术在食品微生物中应用的研究进展

聚合酶链式反应(Polymerase Chain Reaction,PCR)结合变性梯度凝胶电泳(Denaturing Gradient Gel Electrophoresis,DGGE)技术能快速的研究微生物物种多样性,该技术不依赖于微生物的分离培养,便能快速、准确地获取复杂样品中微生物的菌群组成及其遗传信息,因此被广泛应用于微生物生态学、食品微生物学等领域。本文主要介绍了PCR-DGGE在食品微生物中和其他一些方面应用的研究进展。     

浓香型白酒酒醅细菌群落结构及其变化规律

该试验利用PCR-DGGE技术研究了浓香型白酒酒醅细菌群落结构及其发酵过程中的变化规律,结果发现:所有酒醅样品均出现13~23条较清晰的条带,其中第1、2、3、5、6、12、13号条带在所有样品中均有检出,且优势度较高;所有样品生物多样性指数都在2.12~2.80之间,发酵前期,多样性指数总体呈先增加后降低的趋势,发酵第49d时到达最低值,之后多样性指数有所增加,后期保持平稳;相邻发酵时间酒醅细菌DGGE图谱相似性指数较高,为0.58~0.78。     

PCR-DGGE技术及其在云南传统火腿微生物研究中的应用展望

聚合酶链式反应技术(PCR技术)能快速的研究传统发酵肉制品微生物物种的多样性,准确地获取复杂样品中微生物的消长规律而不需要进行微生物的传统培养,因此被广泛应用于各领域的研究。主要介绍了各种PCR-DGGE技术在发酵肉制品微生物方面的应用,并对其在云南传统火腿微生物中的研究应用做出展望。     

应用PCR-DGGE技术分析发酵鸭肉中细菌多样性

对不同前处理的发酵鸭肉应用变性梯度凝胶电泳(DGGE)技术进行细菌菌群分析。以细菌通用引物扩增16S rDNA基因可变区V3区,进行DGGE上样,从而获得可以表征细菌群落结构的图谱。研究结果表明,热炒后发酵和未热炒发酵的2个样品菌群差异比较大,但它们都有乳杆菌(Lactobacillus alimentarius),在发酵过程中乳酸菌是主要优势菌,而且未热炒发酵相对于热炒后发酵的样品各菌群数量比较明显。热炒后发酵的鸭肉中存在乳酸菌属(Lactobacillus sp),赖氨酸芽孢杆菌属(Lysinibacillus sp),乳酸片球菌(Pediococcus acidilactici),葡萄球菌属(Staphylococcus stepanovicii),丛毛单胞菌属(Comamonas sp)。未热炒发酵的鸭肉中有赖氨酸芽孢杆菌属(Lysinibacillus sp),普氏厌氧球菌(Anaerococcus prevotii),戊糖片球菌(Pediococcus pentosaceus),吲哚嗜胨菌(Peptoniphilus indolicus),乳杆菌(Lactobacillus alimentarius)。在发酵鸭肉中还检测到了3种不可培养菌Uncultured Lactobacillus sp、Uncultured bacterium、Uncultured Comamonas sp,这在鸭肉发酵过程中的作用还有待进一步研究和验证。PCR-DGGE能快速地反应发酵鸭肉中细菌群落结构。     

基因组学及其在白酒酿造基础研究领域中的应用

中国白酒酿造属于典型的多菌种固态发酵模式,因特殊的开放式酿造工艺使得参与发酵的微生物种类极其复杂,然而传统分离培养技术限制了对复杂微生物体系的深入研究.基因组学以其通量高、规模大等特点为解析复杂多样的生物体基因组提供了技术手段,能够实现对传统白酒固态发酵微生物及其表达调控的研究,最终揭示微生物与功能代谢物之间的内在关联...     

PCR-DGGE技术对中华开菲尔微生物菌群的分析

为了解中华开菲尔微生物菌群的结构特征,本论文运用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术对开菲尔菌株发酵过程中微生物菌群的结构变化进行了实验分析,结果表明:细菌菌群DGGE图谱上出现有三种不同迁移位置的斑带,而酵母菌群DGGE图谱上只有一条斑带;经过DNA序列的对比分析可知:细菌菌群分别为肠膜明串珠菌(Leuconostoc mesenteroides)、马乳样乳杆菌(Lactobacillus kefiranofaciens)和开菲尔乳杆菌(Lactobacillus kefir),它们的序列同源性都达到100%;酵母菌群为德尔布有孢圆酵母(Torulaspora delbrueckii),其序列同源性为99%。本论文首次报道了德尔布有孢圆酵母在开菲尔菌落中的存在。     

PCR-DGGE指纹分析技术在食品微生物检测中的应用

多聚酶链式反应结合变性梯度凝胶电泳指纹分析技术（PCR-DGGE）常用于分析自然环境中微生物群体的遗传多样性,具有可重复和容易操作且不需要进行微生物培养等特点,近年来该技术开始作为食品微生物的研究手段。本文概述了PCR-DGGE指纹分析技术在国外发酵食品及相关生态环境的研究中的应用现状与前景。      

Bacterial and fungal diversity in the traditional Chinese strong flavour liquor Daqu

In this study, the diversity of bacterial and fungal communities in the strong flavour Daqu production process was investigated using polymerase chain reaction denatured gradient gel electrophoresis (PCR‐DGGE). The band number, dominance, diversity and similarity of the 16S rDNA differed in the DGGE patterns, reflecting the wide diversity of the microbial communities present in the different phases. The band abundance and dominance of the eukaryotic microbial community varied in the different phases. To provide insight into factors contributing to this diversity, the physicochemical properties of Daqu were explored. The core temperature of Daqu gradually increased to a maximum during the Dinghuo phase followed by a decline. The moisture and starch content decreased throughout fermentation. The total acid, reducing sugar, amino nitrogen, α‐amylase activity, glucoamylase activity, protease activity, bacterial and mould number increased gradually to a maximum value at the end of the Peijun phase before decreasing in the Dinghuo phase increasing slightly in the Huanluo phase and then remaining almost unchanged. The esterase activity exhibited a similar change to that of other enzymes and then slightly increased after the Peijun stage. For yeasts, the highest number was obtained at the Peijun stage and then decreased. Correlation analysis revealed that the diversity of both bacteria and fungi was negatively correlated with moisture and starch and positively correlated with other indices. The reducing sugar, amino nitrogen, α‐amylase and protease activities showed significant positive correlation with bacteria, and glucoamylase and esterase showed significant correlation with fungi. © 2019 The Institute of Brewing & Distilling     

Microbial community structure of pit mud in a Chinese strong aromatic liquor fermentation pit

Pits are necessary solid bioreactors for the production of Chinese strong aromatic liquor, and the microbial community in these pits significantly affects the quality and yield of the liquor. In this study, the microbial community structures of different aged pit muds were evaluated by phospholipid fatty acid analysis. The results indicated that the microbial community was composed of bacteria, actinomycetes and fungi, with Gram‐positive (G⁺) bacteria and anaerobic bacteria being dominant. The biomass of the pit mud increased and the microbial community shifted to G⁺ bacteria as the pit age increased. Principal component analysis showed that PC1 consisted mainly of fatty acids of saturated (15:0, 16:0 and 18:0), methyl‐branched (a15:0, i16:0, a16:0, i17:0 and 8Me18:0), polyunsaturated (16:1ω7c, 16:1ω9) and cyclopropane (cy17:0 and cy19:0), and PC2 mainly of 18:1ω9, 18:1ω10 and a14:0, The characteristics of the microbial community structure in the pit mud resulted from the stress effects of low oxygen concentration, low pH and a high alcohol concentration. Copyright © 2013 The Institute of Brewing & Distilling     

中华酒道——品酒之道

中华酒道是酒文化的重要表现形式,它将酒文化由抽象无形进而变为让人们有形可循。品酒是中华酒道系统中重要核心部分,它包括酒的质量水平评价,精神享受愉悦度,食品风味感受度,酒品安全评价,典型风格的辨别,生产的合理性的评价等。只有掌握基本的品酒技巧,才能辨别酒的品味、品质、品格、品种,才能提高酒品质量、勾调技巧、工艺水平,实现科技创新。     

固定化酵母在清香型白酒发酵的应用研究

采用固定化酵母发酵技术对传统白酒生产工艺进行部分改造，对其发酵工艺的研究结果显示，利用固定化酵母的效果均比对照组的产量和出酒率高3％，且不影响原酒的风味和其他理化指标。     

白酒中甜味剂的检测及其来源的探讨

建立了白酒中4种甜味剂（安赛蜜、甜蜜素、糖精钠、三氯蔗糖）的高效液相色谱-串联质谱的检测方法。采用Agilent EC-C18色谱柱,以甲醇-乙酸铵溶液为流动相,柱温35 ℃,流速0.8 mL/min梯度洗脱进行检测。结果表明,4种甜味剂在5～100 ng/mL范围内呈良好线性关系（相关系数R2≥0.998 6）,检出限为0.001～0.02 mg/kg,回收率在82.70%～117.62%之间,相对标准偏差（RSD）为1.09%～4.64%。该检测方法快速灵敏,大大缩短了检测周期和检测成本,并通过大量样品的检测对白酒中甜味剂的来源进行了探讨。     

Microbial succession and metabolite changes during the fermentation of Chinese light aroma‐style liquor

A combination of culture‐dependent and culture‐independent methods and SPME–GC–MS were used to monitor changes of bacterial and yeast communities, and flavour compounds during the fermentation process of Chinese light aroma‐style liquor. Bacillus and Lactobacillus were the main bacterial genera. Pichia anomala, Saccharomyces cerevisiae and Issatchenkia orientalis were the dominant yeast species. There was a close relationship between fermentation time and the shift of microbial community. Compared with the microbiota in the fermentation of other style liquors, higher bacterial diversity and different non‐Saccharomyces composition led to a variety of metabolites. Metabolite analysis showed that esters, acids, alcohols, aromatic compounds and phenols were the main flavour components and most of them were synthesised in the latter phase of fermentation. Principal component analysis further demonstrated that Bacillus and yeast were the most influential microorganisms in the first 10 days of fermentation, and lactic acid bacteria predominated in the later phase. Lactic acid bacteria regulated the composition of other bacteria and yeast, and synthesised flavour compounds to affect the organoleptic properties of liquor. S. cerevisiae and P. anomala were two important yeast species responsible for the characteristic aroma of liquor. These results present a comprehensive understanding of microbial interaction and potential starter cultures to produce desirable liquor quality. © 2018 The Institute of Brewing & Distilling     

三论中华酒道——中国酒酿造技艺、风味之道与保护篇

介绍了中国酿酒技艺之道、酒类勾调技术之道和白酒的风味之道。     

固态白酒蒸馏过程中潽甑现象的实验研究

对固态白酒实际生产装置潽甑操作工艺条件、流酒产量和风味物质组成进行了实验研究.测试分析结果表明,潽甑现象主要发生在流酒之前;潽甑不但降低了蒸馏效率,而且改变了白酒内各种风味物质的含量,严重影响了正品酒的品质.文中结合各风味物质在白酒蒸馏过程中馏出规律,分析了潽甑发生的基本原因,提出了防止潽甑的相应措施,取得良好效果.