초록

III. 연구개발의 내용 및 범위
● E. coli에서 glycosylation 하기 위한 nucleotide-sugar 생합성 유전자 cassette의 제작
● UDP-glucose 및 유도체를 과생산하기 위한 대장균의 유전자 변이
● E. coli 균주에서 glycosylated 생리활성물질의 합성전략

Abstract ▼

Glycosylation of secondary metabolites is one of the most common modifications in the plants and other producers, which confers physical changes including solubility and stability, as well as biological changes (Ruffing and Chen 2006). For example, glycosylation of the therapeutically important comp

Glycosylation of secondary metabolites is one of the most common modifications in the plants and other producers, which confers physical changes including solubility and stability, as well as biological changes (Ruffing and Chen 2006). For example, glycosylation of the therapeutically important compounds erythromycin, amphotericin B, avermectins, doxorubicin, and quercetin is important for the interaction between the drugs and their cellular targets (Méndez and Salas 2001, Salas and Me´ndez 2007). Moreover, glycosylation may improve absorption, distribution, metabolism, and excretion (ADME properties) of drugs (Thorson et al. 2004) and detoxification of exogenous compounds (Vogt and Jones 2000).
Major results are the development of production system of 4-amino-4,6-dideoxygalactosyl flavonoids and the synthesis of 4-aminodideoxy galactoxyl quecetin, kaempferol and ficetin in E. coli. E. coli was engineered to produce TDP-4-aminodideoxy galactose and added in ArGT3 as glycosyltransferase. Several flavonoids were fed in this system, incubated and extracted. The synthesized 4-amino-4,6-dideoxygalactosyl flavonoids were isolated and characterized by LC-Mass.
Also the production system of 3-amino-3,6-dideoxygalactosyl flavonoids were developed and this system used to synthesize 3-O-3-amino-3,6-dideoxygalactosequercetin, ficetin, kaempferol and naringenin. The 3-rhamnosul kaepferol, 3-rhamnosyl-ficetin and 3-rhamnosyl-morin were produced in E. coli BL21(DE3)/Δpgi TGSDH-EPKR-Gt-3 and then characterized by LC-Mass and NMR.
E. coli BL21(DE3)/ΔpgiΔzwfΔushA mutants were constructed to enhance whole cell bioconversion of exogeneously supplied quercetin into 3-O-xylosyl quercetin.
The maximum concentration of 3-O-xylosyl quercetin achieved was 23.78 mg/L (54.75 μM) representing 54.75 % bioconversion, which was ~4.8-fold higher bioconversion than that shown by E. coli BL21 (DE3) with the same set of genes. Bioconversion was further improved by 98% when the reaction was scaled up in a 3 L fermentor at 36 h.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 5
• 목차 ... 7
• 제1장 서 론 ... 8
• 제1절 연구개발의 필요성 ... 8
• 제2절 연구개발대상 기술의 경제적 및 산업적 중요성 ... 10
• 제2장 국내외 기술개발 현황 ... 12
• 제1절 국내 연구 현황 ... 12
• 제2절 국외 연구 현황 ... 12
• 제3장 연구개발수행 내용 및 결과 ... 15
• 제1절 추진전략·방법 ... 15
• 제 2 절 연구결과 ... 17
• 제4장 연구개발목표 달성도 및 대외기여도 ... 37
• 제1절 목표대비 달성도 ... 37
• 제2절 정량적 성과 ... 38
• 제5장 연구개발결과의 활용계획 ... 41
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 42
• 제7장 기타 중요 변동사항 ... 43
• 제8장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 44
• 제9장 참고문헌 ... 45
• <참고 2> 주요 연구 성과 ... 47
• 주요 결과 요약서 ... 49