榨菜发酵过程中原核微生物群落结构及其理化因子的动态演替

利用16S rRNA高通量测序技术探究榨菜发酵过程中原核微生物群落的演替,同时利用物理化学方法追踪发酵过程中理化因子的变化,并进行冗余分析。结果表明,随着榨菜发酵的不断进行,乳杆菌属的相对丰度从0. 92%增加至74. 77%,是发酵过程中的绝对优势属。明串珠菌属在发酵初始时的相对丰度可达3. 61%,在整个发酵过程中呈先增加后降低的变化趋势,可起到启动剂的作用。冗余分析表明:发酵初期的群落与p H及还原糖含量呈正相关;发酵中期的群落与亚硝酸盐、丙酮酸和琥珀酸含量呈正相关;而发酵后期的群落与总酸和其他有机酸含量呈正相关。该研究较系统地揭示了榨菜发酵过程中原核微生物群落的演替规律及其影响因素,可为进一步精确人工调控提供理论依据和筛选特性菌种提供数据参考。     

汉中地区浆水发酵过程中微生物群落演替及其功能预测

该研究利用Illumina MiSeq测序技术,对发酵不同阶段的汉中浆水进行细菌与真菌多样性检测分析。结果表明,随着发酵的进行,细菌群落的丰度和多样性较为稳定,真菌群落的丰度和多样性先增加后趋于稳定。3个发酵时期中优势细菌门均为厚壁菌门（Firmicutes）,优势细菌属为乳植杆菌属（Lactiplantibacillus）、腐败乳杆菌属（Loigolactobacillus）、魏斯氏菌属（Weissella）、寡碳乳杆菌属（Paucilactobacillus）、明串珠菌属（Leuconostoc）等;优势真菌门为担子菌门（Basidiomycota）和子囊菌门（Ascomycota）,优势真菌属主要有双胞菌属（Cystofilobasidium）、白冬孢酵母属（Leucosporidium）、镰刀菌属（Fusarium）、德巴利酵母属（Debaryomyces）、Holtermanniella等。整个发酵过程中优势菌属相对含量有明显波动,参与发酵的真菌种类多于细菌。微生物功能预测结果显示,细菌中涉及代谢、遗传信息处理和环境信息处理的通路比例分别为（74.11±0.11）%、（12....     

黑树莓酒自然发酵过程中微生物群落演替研究

以自然发酵的黑树莓酒为研究对象,利用高通量测序技术探究其自然发酵过程中的微生物群落组成、结构及演替变化规律。结果表明,在酒精发酵（AF）过程中,酵母属（Saccharomyces）、弯担菌属（Curvibasidium）和木拉克酵母属（Mrakia）相对丰度较高,酿酒酵母（Saccharomyces cerevisiae）在中期和后期相对丰度达75%以上。在苹果酸-乳酸发酵（MLF）过程中,泛菌属（Pantoea）、酒球菌属（Oenococcus）、乳球菌属（Lactococcus）、酸土单胞菌属（Aciditerrimonas）和地芽孢杆菌属（Geobacillus）占有较高的比例。酒球菌属增殖迅速,增长了近25%,在苹果酸-乳酸发酵过程中增幅最大。     

传统清香型白酒发酵过程中真菌群落结构及其动态演替

通过Illumina MiSeq高通量测序技术,对传统清香型白酒大楂和二楂发酵过程中的真菌群落结构、丰度及演替规律进行研究,测序获得有效序列数分别为488 956条和642 670条,分别包含14 230个和7 303个OTU,归类为26个亚门或纲,主要为子囊菌纲、担子菌亚门、接合菌亚门和其它未分类的单元。结果表明,传统清香型白酒大楂和二楂真菌群落组成丰富,在发酵过程中真菌的种类和数量随着发酵时间的延长和环境的改变而不断变化,其中子囊菌纲的腹膜孢酵母属、酵母属、假丝酵母属、有孢汉逊酵母属和有孢圆酵母属为传统清香型白酒大楂或二楂发酵主要优势菌群。本研究为清香型白酒发酵过程中功能性微生物菌群的进一步挖掘奠定了基础。     

传统清香型白酒发酵过程中细菌群落结构及其动态演替

利用Illumina MiSeq高通量测序技术,对传统清香型白酒大楂和二楂发酵过程中的细菌群落结构、丰度及演替规律进行研究,测序获得有效序列数分别为319 872条和269 087条,归类为15个门(厚壁菌门、拟杆菌门、变形菌门、放线菌门、浮霉菌门、绿弯菌门、疣微菌门、酸杆菌门、芽单胞菌门、衣原体门、硝化螺旋菌门、广古菌门、产金菌门、梭杆菌门、蓝细菌门,分别包含39 567,22 717个OTUs。研究表明:大楂和二楂细菌群落组成丰富,在发酵过程中细菌的种类和数量随着发酵和环境的变化而变化,主要优势菌群包括葡萄球菌属、片球菌属、乳杆菌属。它们动态变化,此消彼长,影响大楂和二楂酒的质量与风味物质的产生。     

鲟鱼发酵过程中微生物的演替变化分析

为了解鲟鱼发酵中细菌群落多样性及动态演替规律,为发酵鲟鱼的品质提供理论数据,利用Illumina HiSeq测序平台,采集鲟鱼发酵过程中的7个阶段的样品,对样品中的细菌16S rDNA V4~V5区进行测序和系统发育分析。结果表明,样品获得序列数平均为46 463条,各个发酵阶段的操作分类单元数目分别为新鲜鱼样1 064,腌制鱼样952;发酵5 d 454,发酵10 d 442,发酵20 d 327,发酵25 d 372和发酵35 d 356。7组样品中总体细菌组成较为复杂,含有38个门、196个科,在新鲜鱼样中,厚壁菌门(Firmicutes)和变形菌门(Proteobacteria)占主要优势;在腌制阶段,蓝藻门(Cyanobacteria)占主要优势;从发酵开始至结束,厚壁菌门(Firmicutes)占绝对优势。在科水平上,新鲜鱼样中的主要优势菌科为肠杆菌科(Enterobacteriaceae),腌制样品中的主要优势菌科为肠球菌科(Enterococcaceae);从发酵开始至结束,主要的优势菌科为明串珠菌科(Leuconostocaceae),其次是乳杆菌科(Lactobacillaceae)和链球菌科(Streptococceae)。通过HiSeq测序能够更全面解析鲟鱼发酵过程中的细菌多样性;从测序平台提供的微生物多样性信息中发现,各样品间的菌属丰度存在一定的差异性,说明菌群组成与发酵环境密切相关,这为传统腌鱼发酵提供了理论依据和数据支撑。     

基于高通量测序的郫县豆瓣不同发酵期细菌群落结构及其动态演替

采用Illumina MiSeq高通量测序技术,对郫县豆瓣全发酵过程中的细菌群落结构、丰度及演替规律进行深入研究,测序获得有效序列数731188条,归类为11个门（Firmicutes、Proteobacteria、Actinobacteria、Chlorobi、Chloroflexi、Gemmatimonadetes、Fusobacteria、Cyanobacteria、Bacteroidetes、Acidobacteria、Deferribacteres）和其他未分类的单元,包含37082个OTU。研究结果表明郫县豆瓣细菌群落组成丰富,在发酵过程中细菌的种类和数量随着发酵的持续和环境的变化而不断变化,尤其是peijiao样品,其细菌组成的丰度与其他5个时期的样品（BZ1Y、BZ5Y、BZ10Y、HE1Y和HE5Y）差异显著。另外,还有一些未被分类的细菌新物种资源等待深入挖掘。结果揭示了郫县豆瓣中有185个细菌属,其中Staphylococcus、Weissella、Pediococcus、Lactobacillus、Corynebacterium和Bacillus属的细菌是豆瓣发酵的主要优势菌群,伴随于郫县豆瓣的整个发酵过程,对郫县豆瓣的质量与风味产生重要的影响。     

不同地区发酵浆水中微生物群落结构比较及优势菌群的鉴定

浆水作为我国特色发酵食品,具有丰富的营养价值和独特的口感风味。本试验基于高通量测序技术,对甘肃天水、宁夏固原、陕西安康三个不同地区所采集的浆水中微生物群落结构展开研究,并对其中优势菌群进行鉴定,欲探究浆水中微生物群落结构是否具有地域性差异。结果表明,各地区浆水样品中细菌群落结构比较稳定,差异性较小,真菌群落结构稳定性低,各地区、样本之间差异较大。三个地区优势细菌属均为乳酸杆菌属（Lactobacillus）,在各地区样品中相对丰度在72.69%~99.95%之间,宁夏固原地区样品中醋酸杆菌属（Acetobacter,25.24%）、葡糖醋杆菌属（Gluconacetobacter,1.39%）也占有较高丰度。浆水中主要优势真菌属为双足囊菌属（Dipodascus）其相对丰度在40%以上,但不同样品中突出优势菌属差异较大。该研究揭示了陕西安康、甘肃天水、宁夏固原三个地区浆水中微生物群落结构及多样性的异同,表明三地浆水中微生物群落结构存在一定的地域性差异,为稳定浆水产品质量、口感风味及挖掘不同地区浆水中特色微生物资源提供了一定的理论依据。     

洞口酱香酒第一轮次堆积发酵过程中微生物群落结构及其与理化因子关联性分析

为研究贵州天然溶洞洞口酿造酱香酒过程中复杂微生物群落演替,采用高通量测序技术对洞口酱香酒第一轮次堆积发酵酒醅中微生物菌群结构进行分析。共检出20个细菌门和7个真菌门,以及337个细菌属和96个真菌属,其中主导的细菌门和真菌门分别为Firmicutes和Ascomycota,优势细菌属为Lactobacillus、Virgibacillus和Kroppenstedtia等,优势真菌属为Thermomyces、Aspergillus和Saccharomycopsis等。优势细菌属和优势真菌属与理化因子之间的冗余分析结果显示,堆积发酵阶段第0～48 h细菌群落结构受淀粉含量的影响较大,第72～96 h受水分、酸度和温度的影响较大,对堆积发酵结束第120 h影响最大的理化因子是还原糖。而真菌群落结构在前48 h受淀粉含量影响较大,随着发酵的进行,第72～120 h真菌群落结构受温度、水分、酸度和还原糖的影响。除Lactobacillus、Acetobacter、Komagataeibacter和Penicillium外,大部分的优势菌属与水分、温度、酸度和还原糖呈显著负相关。细菌菌群功能预测结...     

基于高通量测序分析复配小曲白酒发酵过程中微生物群落结构及多样性

利用高通量测序对复配小曲清香型白酒酿造过程中微生物多样性及变化规律进行比较分析。结果表明,样品中共检测出35 个门、378 个属的细菌和4 个门、38 个属的真菌。优势菌门为厚壁菌门（Firmicutes）、变形菌门（Proteobacteria）、拟杆菌门（Bacteroidetes）、子囊菌门（Ascomycota）和毛霉菌门（Mucoromycota）。小曲白酒发酵过程中菌群结构不断发生变化,发酵前2 d细菌优势菌属为不明立克次体菌属（unidentified Rickettsiales）,发酵3 d后的绝对优势菌属为乳杆菌属（Lactobacillus）直至发酵结束。真菌优势菌属为伊萨酵母属（Issatchenkia）、根霉属（Rhizopus）、酵母菌属（Saccharomyces）和曲霉属（Aspergilus）。随着发酵时间延长,根霉属和曲霉属相对丰度都在降低,而酵母菌属在发酵第2天后其相对丰度在不断提高并成为优势菌。本研究揭示了复配小曲白酒酿造发酵过程中微生物群落多样性及变化规律,为复配小曲白酒的生产提供理论支持。     

山西陈醋酿造微生物群落结构及其互作网络分析

为揭示山西陈醋酿造过程微生物群落的演替规律和相互作用,采用高通量测序技术对6个不同发酵阶段醋醅样品的微生物群落组成结构和多样性进行分析,通过斯皮尔曼相关系数（SCC）计算得到物种相关系数矩阵后,并利用互作网络数据可视化的软件Cytoscape 3.7.2研究发酵菌群的共存关系网络。结果表明,醋醅中细菌群落多样性远高于真菌群落。醋醅中优势细菌属为乳酸杆菌属（Lactobacillus）和醋酸杆菌属（Acetobacter）,优势真菌属为复膜孢酵母属（Saccharomycopsis）。细菌互作网络分析表明,醋酸杆菌属（Acetobacter）与乳酸杆菌属（Lactobacillus）、魏斯氏菌属（Weissella）具有显著的拮抗性;乳酸杆菌属（Lactobacillus）和魏斯氏菌属（Weissella）存在协同作用。真菌互作网络分析表明,复膜孢酵母属（Saccharomycopsis）与链格孢属（Alternaria）存在显著负相关关系。     

红曲米醋制曲过程中微生物群落演替及其对生化指标的影响

通过高通量测序对红曲米醋红曲制曲过程中微生物群落多样性进行分析,结合红曲的糖化力、液化力等生化指标,采用典范对应分析（CCA）深度解析微生物群落演替对红曲生化指标的影响。结果显示,分别获得16个真菌属菌株和16个细菌属菌株,主要真菌有曲霉属（Aspergillus）（相对丰度为0.36%～66.79%）、酿酒酵母属（Saccharomyces）（相对丰度为14.77%～47.80%）、红曲霉属（Monascus）（相对丰度为11.28%～41.09%）,随着制曲时间增加,优势真菌由Saccharomyces、Monascus逐渐变为Aspergillus;细菌中乳酸菌（Lactobacillus）为优势菌（相对丰度为63.54%～94.98%）,且随着制曲时间的增加,呈先增后降趋势。CCA分析结果表明,真菌中Aspergillus、Penicillium对红曲生化指标的影响起主要作用,细菌属对红曲生化指标影响甚微。     

Changes in microbial community during fermentation of high‐temperature Daqu used in the production of Chinese ‘Baiyunbian’ liquor

Daqu is a fermentation starter used in the production of Chinese liquor. The present study investigated changes in the microbial community during the fermentation of Baiyunbian high‐temperature Daqu during a cycle time of 140 days. Two methods were used, a culture‐dependent method (viable cell counting) and a culture‐independent method (high‐throughput sequencing of bacterial 16S rRNA gene and fungal internal transcribed spacer region). It was found that thermotolerant microorganisms such as Bacillus , Thermomyces and Actinobacteria were dominant in all stages of Daqu fermentation, especially during the high‐temperature period of days 7–38. In the initial 110 days of fermentation, operational taxonomic units and number of bacteria were higher than those of fungi, but the opposite was observed towards the end of fermentation (day 140). In mature Daqu , the predominant bacterial species were Bacillus and unclassified Thermoactinomycetaceae, followed by Thermoactinomyces , Kroppenstedtia and Saccharopolyspora ; the predominant fungi were Aspergillus , unclassified Trichocomaceae, Thermomyces , Rhizopus , Monascus and Candida , among others. It is possible that the addition of mother Daqu , temperature and storage time played critical roles in the microbial composition of Daqu . Collectively, the above findings provide important information that can be used to optimize conditions for large‐scale production of Daqu . Copyright © 2017 The Institute of Brewing & Distilling     

Effects of cellulase‐producing bacteria on bacterial community structure and diversity during fermentation of Chinese liquor grains

To study role of cellulase‐producing bacteria on bacterial community structure during fermentation of Chinese liquor grains, a cellulase‐producing strain called DM‐4 was added to the grains at different levels. The bacterial community structure and diversity were then studied using a high‐throughput sequencing method. Results showed that the bacterial community structure exhibited varied characteristics at different levels of grain fermentation, which amply illustrated that adding cellulase‐producing bacteria to fermenting grains had significant effect on the bacterial community structure. Diversity analysis indicated that abundance and diversity of bacterial community increased significantly by adding 10⁴–10⁶ cfu/g DM‐4 cellulase‐producing bacteria. Also when 0–10⁶ cfu/g of DM‐4 cellulase‐producing bacteria was added, there was significant correlation between the dose of cellulase‐producing bacteria added and bacterial community diversity. The study thus concluded that bacterial community diversity and uniformity increased by adding cellulase‐producing bacteria during fermentation. Copyright © 2017 The Institute of Brewing & Distilling     

基于16S rRNA的徽派腊肉加工过程中微生物群落结构分析

为探究徽派腊肉加工过程中的微生物群落演替规律,采用高通量测序技术对不同加工阶段的徽派腊肉中微生物16S rRNA进行V3～V4区测序,比较不同加工阶段(鲜肉、腌制中期、腌制结束、成熟中期、成熟结束)腊肉中细菌群落结构组成及多样性差异.结果表明:5个加工阶段分别获得80155、80116、80174、80114、8017...     

宣恩火腿发酵过程中表面微生物群落演替规律

为揭示宣恩火腿发酵过程中微生物群落演替规律,运用高通量测序技术分析不同发酵时间火腿表面细菌和真菌群落组成,并对物种相关性和微生物群落功能进行分析.结果表明:宣恩火腿表面细菌群落多样性在发酵过程中持续增加,共检出11个门和96个属;发酵前、中期,木糖葡萄球菌(Staphylococcus xylosus)占绝对优势(>9...     

Microbial community dynamics during the Scamorza Altamurana cheese natural fermentation

The growth dynamics of the natural microbial community responsible for the fermentation of Scamorza Altamurana, a typical Southern Italian cheese made using backslopping, was investigated applying a polyphasic approach combining 1) microbial enumeration with culture media, 2) randomly amplified polymorphic DNA (RAPD) fingerprinting of microbial communities, 3) sequencing of partial 16S ribosomal DNA (rDNA) genes, and 4) physiological tests. Viable cell counts on different culture media showed that the cocci community prevailed during the 18 h of curd fermentation and the 6 d of cheese ripening. RAPD fingerprinting made it possible to isolate 25 different strains identified by 16S rDNA sequencing as belonging to five species of Lactobacillus, three species of Streptococcus, one species of Weissella, and one species of Enterococcus. The physiological analyses of all lactic acid bacteria strains revealed that the isolates belonging to Streptococcus genus were the most acidifying, whereas lactobacilli were most proteolytic. Streptococcus thermophilus C48W and Lactobacillus delbrueckii subsp. bulgaricus B15Z dominated all through the fermentation process. Furthermore, they seemed to be stable in a subsequent whey sample analyzed after 7 mo. The recovery of strains endowed with interesting technological features, such as acidifying and proteolytic activities, and surviving in natural whey could allow the upscaling of cheese processing safeguarding the organoleptic characteristics of Scamorza Altamurana and could possibly improve other fermented dairy products.     

Microbial Ecology of Watery Kimchi

The biochemistry and microbial ecology of 2 similar types of watery (mul) kimchi, containing sliced and unsliced radish and vegetables (nabak and dongchimi, respectively), were investigated. Samples from kimchi were fermented at 4, 10, and 20 °C were analyzed by plating on differential and selective media, high‐performance liquid chromatography, and high‐throughput DNA sequencing of 16S rDNA. Nabak kimchi showed similar trends as dongchimi, with increasing lactic and acetic acids and decreasing pH for each temperature, but differences in microbiota were apparent. Interestingly, bacteria from the Proteobacterium phylum, including Enterobacteriaceae, decreased more rapidly during fermentation at 4 °C in nabak cabbage fermentations compared with dongchimi. Although changes for Proteobacterium and Enterobacteriaceae populations were similar during fermentation at 10 and 20 °C, the homolactic stage of fermentation did not develop for the 4 and 10 °C samples of both nabak and dongchimi during the experiment. These data show the differences in biochemistry and microbial ecology that can result from preparation method and fermentation conditions of the kimchi, which may impact safety (Enterobacteriaceae populations may include pathogenic bacteria) and quality (homolactic fermentation can be undesirable, if too much acid is produced) of the product. In addition, the data also illustrate the need for improved methods for identifying and differentiating closely related lactic acid bacteria species using high‐throughput sequencing methods.     

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