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Isolation and Characterization of Lactobacillus plantarum strains with Cholesterol-lowering Activity : 콜레스테롤 저하 활성을 갖는 Lactobacillus plantarum 균주의 분리 및 특성 확인
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 허철성 | - |
| dc.contributor.author | 주동영 | - |
| dc.date.accessioned | 2022-04-05T06:48:41Z | - |
| dc.date.available | 2022-04-05T06:48:41Z | - |
| dc.date.issued | 2021 | - |
| dc.identifier.other | 000000167408 | - |
| dc.identifier.uri | https://hdl.handle.net/10371/177861 | - |
| dc.identifier.uri | https://dcollection.snu.ac.kr/common/orgView/000000167408 | ko_KR |
| dc.description | 학위논문(석사) -- 서울대학교대학원 : 국제농업기술대학원 국제농업기술학과, 2021.8. 주동영. | - |
| dc.description.abstract | Cardiovascular disease is one of the leading causes of death among adults worldwide, and probiotics are continuously being studied as a treatment for cardiovascular disease. Previous studies have demonstrated that probiotics have cholesterol-lowering activities through various mechanisms. The purpose of this study is to screen lactic acid bacteria (LAB) with cholesterol-lowering ability as novel probiotics candidate. In this study, 60 lactic acid bacteria were isolated from 12 fermented foods and infant fecal samples. Compared with commercial probiotics Lactobacillus acidophilus La-5 and Lactobacillus acidophilus NCFM strains, Lactobacillus plantarum DY41 and Lactobacillus plantarum DY42 strains were isolated with high cholesterol assimilation ability. Probiotics properties of the selected strains were evaluated for intestinal viability, epithelial cell adhesion, stability, safety, and antibiotic susceptibility. As a result of in vitro hepatocyte (HepG2 cell line) approach with supernatant of the selected strains, cholesterol accumulation was significantly reduced. Furthermore, in case of the Lactobacillus plantarum DY41 strain, cholesterol incorporation into the cell membrane was significantly increased compared with the Lactobacillus acidophilus La-5 strain. In the Lactobacillus plantarum DY42 strain, the mRNA expression of cholesterol and bile acid-related genes was regulated to cholesterol emission compared to the control. In conclusion, we suggest that the Lactobacillus plantarum strains could be potential probiotic candidates for reducing the risk of developing cardiovascular disease. | - |
| dc.description.abstract | 심혈관 질환은 전 세계 성인 사망의 주요 원인 중 하나이며, 심혈관 질환 치료제로 프로바이오틱스가 지속적으로 연구되고 있다. 이전 연구를 통해, 프로바이오틱스가 다양한 메커니즘을 통해 콜레스테롤 저하 작용을 한다는 사실이 입증되었다. 이 연구의 목적은 새로운 프로바이오틱스 후보로 콜레스테롤 저하 활성을 갖는 유산균 (LAB)을 선발하는 것이며, 12 개의 발효 식품과 유아 분변 샘플에서 60 개의 유산균을 분리했다. 상용 프로바이오틱스인 Lactobacillus acidophilus La-5 및 Lactobacillus acidophilus NCFM 균주와 비교하였을 때, 콜레스테롤 동화 능력이 우수하다고 판단되는 Lactobacillus plantarum DY41 및 Lactobacillus plantarum DY42를 선발하였다. 선택된 균주의 프로바이오틱스 특성은 장 생존력, 상피 세포 부착, 안정성, 안전성 및 항생제 감수성에 대해 평가되었다. 선발된 균주의 supernatant를 사용한 in vitro 간세포 (HepG2 cell line)에 처리한 결과, 콜레스테롤 축적이 현저하게 감소하였다. 또한 Lactobacillus plantarum DY41 균주의 경우, Lactobacillus acidophilus La-5 균주와 비교하여 세포막으로의 콜레스테롤 혼입이 유의하게 증가하였다. Lactobacillus plantarum DY42 균주는 대조군과 비교하여 콜레스테롤 및 담즙산 관련 유전자의 mRNA 발현이 조절되었다. 결론적으로, Lactobacillus plantarum 균주가 심혈관 질환 발병 위험을 줄이는 잠재적인 프로바이오틱스 후보가 될 수 있음을 확인하였다. | - |
| dc.description.tableofcontents | 1 Introduction 1
2 Review of Literature 3 2.1 Cholesterol 3 2.1.1 Cholesterol 3 2.1.2 Physiology 4 2.1.3 Biosynthesis 5 2.1.4 Regulation of synthesis 7 2.1.5 Transport and absorption 9 2.1.6 Recycling and excretion 12 2.2 Dyslipidemia 13 2.2.1 Hypercholesterolemia 13 2.2.2 Hypocholesterolemia 15 2.3 Probiotics and intestinal cholesterol metabolism 16 2.3.1 Probiotics 16 2.3.2 Metabolism of cholesterol by the gut microbiota 19 2.3.3 Metabolism of bile acids by the gut microbiota 21 2.3.4 Genes related to cholesterol and bile acids 22 3 Materials and methods 25 3.1 Screening of putative probiotics strains 25 3.1.1 Isolation of lactic acid bacteria from various samples 25 3.1.2 Determining of probiotic cholesterol assimilation ability in MRS broth 26 3.1.3 16s rRNA sequencing 28 3.1.4 Simulated stomach duodenum-passage (SSDP) 29 3.1.5 Autoaggregation assay 30 3.1.6 Carbon source utilization pattern and phenotypical profiling using API kit / GEN Ⅲ MicroPlate 31 3.1.7 Biogenic amine (BA) production assay 32 3.1.8 Hemolytic activity 34 3.1.9 Antibiotic susceptibility 35 3.2 In vitro evaluation of the selected strains with cholesterol-lowering activity 36 3.2.1 Determination of bile salts hydrolases (BSH) activity 36 3.2.2 Cholesterol assimilation in cellular fractions 38 3.2.3 Incorporation of cholesterol into the cellular membrane 39 3.2.4 Cholesterol assimilation in sterilized whole milk as food matrix 40 3.2.5 Cell culture 41 3.2.6 Intracellular cholesterol accumulation in HepG2 cell line 42 3.2.7 Quantification of mRNA expression in HepG2 cell line 43 3.3 Reference strains 45 3.4 Statistical analysis 45 4 Results 46 4.1 Screening of putative probiotics strains 46 4.1.1 Determination of probiotic cholesterol assimilation ability in MRS broth 46 4.1.2 Acid and bile tolerance 48 4.1.3 Autoaggregation 50 4.2 In vitro evaluation of the selected strains with cholesterol-lowering activity 58 4.2.1 Deconjugation of bile salts 58 4.2.2 Cholesterol assimilation in cellular fractions 60 4.2.3 Incorporation of cholesterol into the cellular membrane 62 4.2.4 Cholesterol assimilation in sterilized whole milk as food matrix 63 4.2.5 Intracellular cholesterol accumulation in HepG2 cell line 64 4.2.6 Quantification of mRNA expression in HepG2 cell line 66 5 Discussion 68 6 References 81 Abstract in Korean 109 | - |
| dc.format.extent | xii, 111 | - |
| dc.language.iso | eng | - |
| dc.publisher | 서울대학교 대학원 | - |
| dc.subject | probiotics | - |
| dc.subject | Lactobacillus plantarum | - |
| dc.subject | cholesterol-lowering | - |
| dc.subject | 프로바이오틱스 | - |
| dc.subject | 콜레스테롤 저하 | - |
| dc.subject.ddc | 631 | - |
| dc.title | Isolation and Characterization of Lactobacillus plantarum strains with Cholesterol-lowering Activity | - |
| dc.title.alternative | 콜레스테롤 저하 활성을 갖는 Lactobacillus plantarum 균주의 분리 및 특성 확인 | - |
| dc.type | Thesis | - |
| dc.type | Dissertation | - |
| dc.contributor.AlternativeAuthor | CHOO Dongyoung | - |
| dc.contributor.department | 국제농업기술대학원 국제농업기술학과 | - |
| dc.description.degree | 석사 | - |
| dc.date.awarded | 2021-08 | - |
| dc.contributor.major | 경제동물과학전공 | - |
| dc.identifier.uci | I804:11032-000000167408 | - |
| dc.identifier.holdings | 000000000046▲000000000053▲000000167408▲ | - |
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