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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 許輔(Fuu Sheu) | |
| dc.contributor.author | I-Hsuan Lo | en |
| dc.contributor.author | 羅逸軒 | zh_TW |
| dc.date.accessioned | 2022-11-25T06:33:32Z | - |
| dc.date.copyright | 2021-08-18 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-07-14 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/82198 | - |
| dc.description.abstract | 台式泡菜常於製作過程添加食醋及菌酛 (starter)，以加速並穩定發酵過程。傳統上以前次發酵完成之泡菜汁液，作為後一批次發酵之循環菌酛 (backslopping fermentation)。另一方面，總基因體學 (metagenomics) 結合次世代定序技術，可分析環境中生物多樣性及菌相消長。本研究以總基因體學為基礎之非經培養 (culture-independent) 方法，分析台式泡菜發酵過程各階段菌相及產物變化，並探討循環菌酛及食醋於其中扮演的角色。比較添加及未添加菌酛之組別，添加菌酛組 (R2) 於發酵四天後，可滴定酸度，與未添加菌酛組 (R1) 發酵五天相當，約為 0.63%。以real-time PCR對16S rRNA基因複製數 (copy number) 進行定量，R2組之泡菜，於發酵第二天總菌含量達到9.0 log copies mL-1以上，而R1組則需要三天才能達到。菌相組成則以高通量擴增子定序分析泡菜總基因體中 16S rRNA 基因V3-V4片段，結果顯示R1組菌種組成相對複雜，而R2組則有穩定一致的菌相。R1組之台式泡菜優勢菌屬從發酵第二天至第四天依序為 Lactococcus、Leuconostoc、Lactobacillus，其相對含量分別為53%、58%、85%；R2組之泡菜菌相並無消長情形，Lactobacillus為優勢菌屬，其中之優勢菌種為Lactobacillus plantarum，並有未經培養的 Lactobacillus 及 Leuconostoc 菌種存在。添加食醋製作之泡菜 (vR2) 與未添加食醋組 (nvR2) 相比，Lactobacillus 於發酵 18 小時後成為優勢菌屬，且在2% (w/w)鹽濃度下，可於發酵 24 小時內抑制非乳酸菌之細菌生長，顯示於低鹽濃度下添加食醋，可快速促進乳酸菌成為泡菜之優勢菌。本研究建立以總基因體學為基礎之微生物組成分析平台，相較於傳統微生物培養之方法，具有高效且完整之優勢；並以此平台深入分析樣品菌相至菌種及菌屬層面，說明添加循環菌酛與食醋可穩定發酵過程之菌相，同時找到多種於發酵製程穩定存在而未被培養之乳酸菌，此平台未來可用於監測食品加工過程之菌相變化並鑑別其中未經培養之重要微生物。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-25T06:33:32Z (GMT). No. of bitstreams: 1 U0001-1207202117053400.pdf: 3941217 bytes, checksum: 5f7f12ffc645697b43963c151b6e278d (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | "致謝 I 摘要 II ABSTRACT III CONTENT V LIST OF TABLES VIII LIST OF FIGURES X ABBREVIATION LIST XII CHAPTER 1 INTRODUCTION 1 1.1 Metagenomics and its methodology 1 1.1.1 Methodology of metagenomics in the past 1 1.1.2 Methodology of metagenomics nowadays 2 1.1.3 Next generation sequencing 3 1.2 Methodology for characteristics of microbiota 4 1.2.1 Culture-dependent method 4 1.2.2 Culture-independent method based on 16S rRNA gene 5 1.2.3 16S rRNA gene amplicon sequencing 6 1.2.4 16S rRNA gene quantification 8 1.3 Taiwanese-style pickle 9 1.3.1 Starter added cultivation 12 1.4 The aim of this study 13 CHAPTER 2 MATERIALS AND METHODS 15 2.1 Chemicals and reagents 15 2.2 Taiwanese-style pickle preparation 15 2.3 Sensory evaluation 16 2.4 Acidity analysis 16 2.5 Total bacterial genomic DNA purification for 16S rRNA gene analysis 16 2.6 Bacterial 16S rRNA gene copy number analysis by real-time PCR 17 2.7 16S rRNA gene amplicon sequencing by next generation sequencing 18 2.8 Next generation sequencing data analysis 19 2.9 Sample derivatization for GC-MS 19 2.10 Gas chromatography coupled with mass spectrometry 20 2.11 Statistical analysis 21 CHAPTER 3 RESULTS 22 3.1 Determination of the fermentation conditions 22 3.2 The titratable acidity of Taiwanese-style pickle during fermentation 23 3.3 Quantification of total bacteria, lactic acid bacteria, and yeast 24 3.3.1 Quantification of total bacteria 25 3.3.2 Quantification of total lactic acid bacteria 25 3.3.3 Quantification of total yeast 26 3.4 Sequencing of 16S rRNA gene amplicons from Taiwanese-style pickle samples 26 3.5 Microbiota diversity of Taiwanese-style pickle samples 27 3.5.1 Alpha diversity 27 3.5.2 Beta diversity 28 3.6 Relative abundance and succession of microbiota in R1 and R2 28 3.7 Species composition of Lactobacillus and Leuconostoc 29 3.8 Determination of metabolites relative abundance 30 3.9 Role of vinegar in the starter added fermentation 31 3.9.1 Summary of NGS results for vR2 and nvR2 32 3.9.2 Diversity of vR2 and nvR2 Taiwanese-style pickle 32 3.9.3 Bacterial genera composition of vR2 and nvR2 33 CHAPTER 4 DISCUSSION 35 4.1 Culture-independent approach compared with traditional culture method 35 4.2 Metagenomics strategy reveals critical role of starter addition in fermentation 36 4.3 Role of vinegar in Taiwanese-style pickle production 37 4.4 Conclusion 39 REFERENCE 40 TABLES 54 FIGURES 66 " | |
| dc.language.iso | en | |
| dc.subject | 菌相 | zh_TW |
| dc.subject | 微生物定量 | zh_TW |
| dc.subject | 非經培養法 | zh_TW |
| dc.subject | 菌酛 | zh_TW |
| dc.subject | 食醋 | zh_TW |
| dc.subject | vinegar | en |
| dc.subject | culture-independent method | en |
| dc.subject | bacterial community | en |
| dc.subject | quantification of microorganisms | en |
| dc.subject | starter | en |
| dc.title | 以總基因體學探討台式泡菜發酵之研究 | zh_TW |
| dc.title | A comprehensive study of Taiwanese-style pickle fermentation using metagenomic strategy | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蘇南維(Hsin-Tsai Liu),周志輝(Chih-Yang Tseng),官彥州,鄭光成 | |
| dc.subject.keyword | 非經培養法,菌相,微生物定量,菌酛,食醋, | zh_TW |
| dc.subject.keyword | culture-independent method,bacterial community,quantification of microorganisms,starter,vinegar, | en |
| dc.relation.page | 83 | |
| dc.identifier.doi | 10.6342/NTU202101413 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2021-07-15 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 園藝暨景觀學系 | zh_TW |
| dc.date.embargo-lift | 2024-07-12 | - |
| 顯示於系所單位： | 園藝暨景觀學系 | |
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