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Isolation and Characterization of a Mucin-Degrading Bacterium Attached to the Rumen Epithelium of Korean Cattle
한우의 반추위벽으로부터 분리한 뮤신 분해 박테리아의 배양 및 특성 규명

DC Field Value Language
dc.contributor.advisor백명기-
dc.contributor.author나상원-
dc.date.accessioned2018-12-03T01:45:48Z-
dc.date.available2020-01-02T23:58:03Z-
dc.date.issued2018-08-
dc.identifier.other000000152546-
dc.identifier.urihttps://hdl.handle.net/10371/143938-
dc.description학위논문 (석사)-- 서울대학교 대학원 : 농업생명과학대학 농생명공학부, 2018. 8. 백명기.-
dc.description.abstractThis study was performed to isolate mucin degrading bacteria attached to rumen epithelial cells. Microbial diluent was prepared by extensive homogenization of rumen epithelial cells of the ventral sacs of Korean cattle steers, and it was inoculated with a liquid medium containing a mucin as a sole carbon source in anaerobic condition. Enriched bacterial culture was serially diluted and spread on a mucin agar medium. Single colonies were picked and cultured in the same liquid mucin medium.

A novel bacterium, designated strain E39T, was isolated and its 16S rRNA gene was sequenced. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain E39T was most closely related to Alloprevotella rava with an 87.3% sequence similarity. Strain E39T is Gram-negative, strictly anaerobic, and non-motile coccus. Growth of strain E39T was observed at 30–45°C (optimum, 39°C), at pH 6.5 and 8.5 (optimum, 7.5), and in the presence of 0.0–1.0% (w/v) NaCl (optimum, 0.0–0.5%). The G+C content of genomic DNA was 46.2 mol%. Enzyme profiles were identified with the Rapid ID 32A identification kit (bioMe´rieux), and this species had was arginine dihydrolase, α-galactosidase, β-galactosidase, glutamic acid decarboxylase, α-fucosidase, alkaline phosphatase, leucyl glycine arylamidase, alanine arylamidase, and glutamyl glutamic acid arylamidase activity. On the basis of the phenotypic, chemotaxonomic, and physiological properties, strain E39T represents a novel genus of the family Prevotellaceae
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dc.description.abstractthe name Psuedoprevotella muciniphila gen. nov., sp. nov. is proposed-
dc.description.abstractthe type strain is E39T (= KCTC 15717T= JCM 32621T).-
dc.description.tableofcontentsⅠ. Introduction 1

Ⅱ. Literature review 3

1. Rumen microbiology 3

1.1. Culture-dependent studies 3

1.2. Culture-independent studies 5

2. Rumen epithelial bacteria 9

2.1. Rumen epithelium and epimural community 10

2.2. Diversity of the epimural community 10

2.3. Functions of the epimural community 13

2.3.1. Tissue recycling 13

2.3.2. Oxygen scavenging 13

2.3.3. Digestion of urea 14

3. Mucin and mucin-degrading bacteria 15

3.1. Mucus and mucin 15

3.2. Mucin-degrading bacteria 16

Ⅲ. Materials and methods 18

1. Enrichment and isolation of mucin-degrading bacteria from the rumen epithelium tissue 18

1.1. Media preparation 18

1.2. Sample preparation and enrichment of mucin-degrading bacteria 19

1.3. Isolation of mucin-degrading bacteria 20

2. Identification of mucin-degrading bacteria based on 16S rRNA gene sequence 20

2.1. DNA extraction and 16S rRNA gene amplicon PCR 20

2.2. 16S rRNA gene sequencing and comparative analysis 21

3. Characterization of strain E39T, a novel mucin-degrading bacterium attached to the rumen epithelium 21

3.1. Analysis of 16S rRNA gene squence and construction of phylogenetic tree 22

3.2. Analysis of cell morphology and Gram staining test 22

3.3. Growth test 23

3.4. Oxygen tolerance test 25

3.5. Enzyme profile test and analysis of cellular fatty acid composition 25

3.6. Whole genome sequencing 26

Ⅳ. Results 27

1. Isolation and identification of mucin-degrading bacteria from the rumen epithelium tissue 27

2. Characterization of strain E39T, a novel mucin-degrading bacterium attached to the rumen epithelium 28

2.1. Analysis of 16S rRNA gene squence and construction of phylogenetic tree 28

2.2. Phenotypic and growth characteristics of strain E39T 33

2.3. Enzyme profile of strain E39T 36

2.4. Cellular fatty acid composition of strain E39T 38

2.5. Whole genome sequencing 40

Ⅴ. Discussion 42

Ⅵ. Literature cited 47

Ⅶ. Summary in Korean 62

Ⅷ. Acknowledgement 64
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dc.formatapplication/pdf-
dc.format.mediumapplication/pdf-
dc.language.isoko-
dc.publisher서울대학교 대학원-
dc.subject.ddc630-
dc.titleIsolation and Characterization of a Mucin-Degrading Bacterium Attached to the Rumen Epithelium of Korean Cattle-
dc.title.alternative한우의 반추위벽으로부터 분리한 뮤신 분해 박테리아의 배양 및 특성 규명-
dc.typeThesis-
dc.contributor.AlternativeAuthorSang Weon Na-
dc.description.degreeMaster-
dc.contributor.affiliation농업생명과학대학 농생명공학부-
dc.date.awarded2018-08-
Appears in Collections:
College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Agricultural Biotechnology (농생명공학부)Theses (Master's Degree_농생명공학부)
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