Process development of producing fucosylated oligosaccharides from colanic acid by engineered Saccharomyces cerevisiae
- 주제(키워드) Fucose , Colanic acid , Acid hydrolysis , 2'-Fucosyllactose , 2'-Fucosylgalactose
- 발행기관 고려대학교 대학원
- 지도교수 김경헌
- 발행년도 2022
- 학위수여년월 2022. 8
- 학위명 석사
- 학과 및 전공 대학원 생명공학과
- 세부전공 시스템식품생명공학과
- 세부분야 해당없음
- 원문페이지 66 p
- 실제URI http://www.dcollection.net/handler/korea/000000268906
- UCI I804:11009-000000268906
- DOI 10.23186/korea.000000268906.11009.0001384
- 본문언어 영어
초록/요약
Fucose is a rare sugar obtained from fucoidan in brown algae. However, fucoidan contains low amounts of fucose, making the process of obtaining fucose inefficient and expensive. Colanic acid (CA) is a capsular exopolysaccharide produced by Escherichia coli that has great potential as a source of fucose owing to its high proportion of fucose. In this study, I aimed to efficiently obtain fucose from the hydrolysis of CA and then produce 2´-fucosyllactose (2´-FL) and 2´-fucosylgalactose (2´-FG) using fucose in the CA using engineered yeast. For the acid hydrolysis of CA, operating conditions of 110°C, 4% (w/v) H3PO4, and 60 min were used. Using fucose from CA hydrolysate as a substrate, 2´-FL and 2´-FG were produced from engineered Saccharomyces cerevisiae. This is the first report of producing fucosylated oligosaccharides from CA. Considering the benefits of CA in obtaining fucose, these results can lead to further applications of CA as a fucose supplier.
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ABSTRACT OF THE THESIS i
LIST OF FIGURES v
LIST OF TABLES vi
1. Introduction 1
2. Materials and Methods 6
2.1. Mass production of colanic acid 6
2.2. Quantitative determination of colanic acid 8
2.3. Compositional analysis of colanic acid 10
2.4. Acid hydrolysis of colanic acid to produce fucose-containing hydrolysate 11
2.5. 2´-Fucosyllactose production using fucose-containing colanic acid hydrolysate 14
2.6. 2´-Fucosylgalactose production using fucose-containing colanic acid hydrolysate 16
2.7. Analysis of fucosylated oligosaccharides production using gas chromatography/time-of-flight‒mass spectrometry 22
2.8. Production and purification of 2´-fucosylgalactose by gel-filtration chromatography and high-performance liquid chromatography 23
3. Results and Discussion 25
3.1. Compositional analysis of colanic acid 25
3.2. Effect of operating conditions on acid hydrolysis of colanic acid 27
3.3. Production of 2´-fucosyllactose using fucose-containing colanic acid hydrolysate 34
3.4. Production of 2´-fucosylgalactose using fucose-containing colanic acid hydrolysate 39
4. Conclusions 48
References 49
