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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳明汝(Ming-Ju Chen) | |
| dc.contributor.author | Chien-Jang Huang | en |
| dc.contributor.author | 黃千讓 | zh_TW |
| dc.date.accessioned | 2021-06-17T02:34:25Z | - |
| dc.date.available | 2021-08-24 | |
| dc.date.copyright | 2017-08-24 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-08-17 | |
| dc.identifier.citation | References
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68768 | - |
| dc.description.abstract | 「益生菌」是一群在宿主體內達到一定數量時對宿主的健康有益處的微生物。乳酸菌一般被認為是安全的(Generally recognized as safe, GRAS)，和傳統發酵食品，尤其是傳統發酵乳製品，有密切的關係。數百年來，蒙古地區的遊牧民族持續的製作各種發酵乳製品，如Airag（發酵馬乳）和Tarag（發酵之牛、山羊、犛牛乳）等等。為了要從這些傳統發酵乳中篩選新的潛力益生菌株，乳酸菌株的收集就變得非常重要。本研究中使用塗盤培養以及分子生物學的技術篩選蒙古傳統發酵乳中具潛力益生菌能力的乳酸菌株。
依據Enterobacterial Reptitive Intergenic Consensus PCR (ERIC-PCR)的結果為基礎，從106個分離自菌落中歸類出40個獨立菌株。透過16s rRNA以及持家基因(housekeeping gene)中的pheS 和 rpoA兩種基因的鑑定，這些菌株分屬八個菌種：Lactobacillus crustorum、Lactobacillus diolivorans、Lactobacillus kefiri、Lactobacillus paracasei、Lactobacillus plantarum subsp. plantarum、Lactococcus. lactis、Leuconostoc lactis以及Leuconostoc pseudomensenteroides。其中Lactobacillus kefiri為最主要的菌種（24株），Lactobacillus crustorum為排第二的菌種（6株）。 接著，本研究針對低pH值、含膽鹽環境的生存耐受性試驗、胞外多醣（EPS）生產量進行測試並作為挑選潛力益生菌能力的項目。結果顯示，Lactobacillus crustorum、Lactobacillus diolivorans、Lactobacillus kefiri在低pH值（pH 3.0）、含膽鹽（0.3% bile）環境具有較佳的生存能力，且Lactobacillus crustorum和Lactobacillus kefiri同時也具有較好的胞外多醣（EPS）生產量。再經過其他條件的測試，兩株Lactobacillus crustorum的菌株（MCC0017及MCC0029）被預期為新的潛力益生菌株。 | zh_TW |
| dc.description.abstract | “Probiotics” are live microorganisms that provide health benefits on the host, when administered in adequate amounts. Lactic acid bacteria (LAB) are regarded as “Generally recognized as safe (GRAS)”, and are mainly involving in the fermentation of traditional fermented foods, especially in fermented milk products. For centuries, the nomadic peoples of Mongolia have been producing various kinds of traditional fermented milk products such as Airag (fermented mare’s milk) and Tarag (fermented milk of cows, goats and yaks). To explore the potential probiotic strains, it is very important to develop the culture collection at the first step. In this study, culture- and molecular-based methods were used to investigate the potential LAB probiotic strains in the traditional Mongolian fermented milk products.
Based on Enterobacterial Reptitive Intergenic Consensus PCR (ERIC-PCR) profiles, a total of 106 isolates isolated from five Mongolian traditional fermented milk products were categorized into 40 different strains, and identified as belonging to 8 species (Lactobacillus crustorum, Lactobacillus diolivorans, Lactobacillus kefiri, Lactobacillus paracasei, Lactobacillus plantarum subsp. plantarum, Lactococcus. lactis, Leuconostoc lactis, and Leuconostoc pseudomensenteroides) by 16S rRNA and housekeeping gene (pheS and rpoA) sequencing. Lactobacillus kefiri was isolated as the predominant species (24 strains), followed by Lb. crustorum (6 strains). By the screening tests, survival in low pH and bile salt and production of exopolysaccharides (EPS) were characterized as the potential probiotic abilities. As a result, almost of the strains in Lb. crustorum, Lb. diolivorans, and Lb. kefiri showed high survival ability in artificial gastric (pH 3.0) and intestinal (0.3% bile) juices. And, almost of the strains in the Lb. crustorum and Lb. kefiri species showed high ability to produce EPS. Finally, features of survival abilities in low pH and bile salt, and EPS production demonstrated that the two Lb. crustorum strains (MCC0017, MCC0029) could be novel prospective probiotics. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T02:34:25Z (GMT). No. of bitstreams: 1 ntu-106-R04626014-1.pdf: 7479591 bytes, checksum: bbc7624cffdb0ec5ff99d23109f482c7 (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | Content
Abstract in Chinese .......................................................................................................i Abstract in English .....................................................................................................iii Preface ..........................................................................................................................v 1. Literature review .....................................................................................................1 1.1. Probiotics ........................................................................................................1 1.1.1. Definition of probiotics ........................................................................1 1.1.2. Health benefits of probiotics ................................................................1 1.1.3. Source of probiotics .............................................................................2 1.2. Mongolian traditional fermented dairy products .............................................3 1.2.1. Airag ....................................................................................................3 1.2.2. Tarag ....................................................................................................3 1.2.3. LAB in Mongolian traditional fermented dairy products ......................4 1.3. Lactic acid bacteria ..........................................................................................5 1.3.1. Classification of LAB ...........................................................................5 1.3.1.1. Lactobacillus .............................................................................6 1.3.1.2. Lactococcus ..............................................................................6 1.3.1.3. Leuconostoc ..............................................................................7 1.3.1.4. Streptococcus ............................................................................7 1.4. Techniques for typing and identification of LAB ...........................................8 1.4.1. Phenotypic method ...............................................................................8 1.4.2. Genotypic method ................................................................................9 1.4.2.1. Restriction fragment length polymorphism (RFLP) ..................9 1.4.2.2. Random amplified polymorphic DNA (RAPD) .....................10 1.4.2.3. Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) ......................................................................................10 1.4.2.4. Polymerase chain reaction−denaturing gradient gel electrophoresis (PCR-DGGE) .........................................................10 1.4.3. Target gene for identification .............................................................11 1.4.3.1. 16S ribosomal RNA gene ........................................................11 1.4.3.2. Housekeeping gene .................................................................11 1.5. Stresses in the intestinal environment for probiotics .....................................12 1.6. Exopolysaccharides (EPS) ............................................................................13 1.6.1. Classification and composition of EPS ...............................................13 1.6.1.1. Homopolysaccharides .............................................................13 1.6.1.2. Heteropolysaccharides ............................................................14 1.6.2. Bioactivities of EPS ...........................................................................14 1.6.3. Factors influencing on EPS production ..............................................14 1.6.3.1. Temperature ............................................................................14 1.6.3.2. pH value ..................................................................................15 1.6.3.3. Culturing period ......................................................................15 1.6.3.4. Carbon source .........................................................................15 1.7. Aims of my study ..........................................................................................17 2. Materials and methods ...........................................................................................26 2.1. Sampling of traditional fermented milk products ..........................................26 2.2. Isolation of lactic acid bacterial (LAB) strains ..............................................26 2.3. DNA extraction .............................................................................................27 2.4. ERIC-PCR typing .........................................................................................28 2.5. PCR amplification of 16S rRNA and housekeeping gene ..............................28 2.6. 16S rRNA and housekeeping genes sequencing and phylogenetic analyses .29 2.7. Tolerance of LAB strains to low pH and bile salt ..........................................30 2.8. Tolerance assay in artificial gastric and small intestinal juices ......................31 2.9. Long-term tolerance of LAB in artificial gastric and small intestinal juices ..31 2.10. Determination of total viable LAB cell counts ............................................32 2.11. Screening for EPS-producing LAB strains ..................................................32 2.12. Quantitative analysis of produced EPS ........................................................33 2.13. Statistical analysis .......................................................................................33 3. Results .....................................................................................................................37 3.1. Identification of LAB strains .........................................................................37 3.2. Tolerance of LAB strains in artificial gastric and intestinal juices .................38 3.3. Long-term tolerance of LAB strains in artificial gastric and intestinal juices.38 3.4. Screening for EPS producing LAB strains ....................................................39 4. Discussion ...............................................................................................................77 4.1. Isolation and identification of LAB strains from Mongolian traditional fermented dairy products ............................................................................77 4.2. Tolerance assay of LAB strains in artificial gastric and intestinal juices........78 4.3. Screening of LAB strains producing large amount of EPS ............................79 5. Conclusions .............................................................................................................84 References ..................................................................................................................85 | |
| dc.language.iso | en | |
| dc.subject | 傳統發酵乳 | zh_TW |
| dc.subject | 益生菌 | zh_TW |
| dc.subject | 乳酸菌 | zh_TW |
| dc.subject | 耐受性 | zh_TW |
| dc.subject | 胞外多醣 | zh_TW |
| dc.subject | Lactic acid bacteria | en |
| dc.subject | Traditional fermented milk | en |
| dc.subject | Probiotics | en |
| dc.subject | Exopolysaccharides | en |
| dc.subject | Tolerance | en |
| dc.title | 傳統蒙古發酵乳製品中乳酸菌之分離及分子鑑定研究 | zh_TW |
| dc.title | Isolation and molecular-identification of lactic acid bacteria from traditional Mongolian fermented milk products | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.coadvisor | 渡邊幸一(Koichi Watanabe) | |
| dc.contributor.oralexamcommittee | 廖啟成(Chii-Cherng Liao),曾浩洋,陳希嘉(Hsi-Chia Chen) | |
| dc.subject.keyword | 傳統發酵乳,益生菌,乳酸菌,耐受性,胞外多醣, | zh_TW |
| dc.subject.keyword | Traditional fermented milk,Probiotics,Lactic acid bacteria,Tolerance,Exopolysaccharides, | en |
| dc.relation.page | 93 | |
| dc.identifier.doi | 10.6342/NTU201703501 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2017-08-18 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 動物科學技術學研究所 | zh_TW |
| 顯示於系所單位： | 動物科學技術學系 | |
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