엔테로코커스 페컬리스 유래의 독성인자의 동정과 응용: 특성, 고정화, 하이드로젤 형성
Identification and Application of a Virulence Factor (EfEstA) from Enterococcus faecalis: Characterization, Immobilization, and Hydrogel Formation
- 주제(키워드) virulence factor , CLEAs , Hydrogel
- 발행기관 아주대학교
- 지도교수 김두헌
- 발행년도 2013
- 학위수여년월 2014. 2
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 파일정보 pdf
- 실제URI http://www.dcollection.net/handler/ajou/000000016376
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The virulence factors from pathogenic bacteria are useful for industrial applications because of their high stabilities and catalytic properties under harsh conditions. Here, characterization, immobilization, and hydrogel formation of a virulence factor, namely, EfEstA from Enterococcus faecalis are presented. The molecular characteristics of EfEstA were investigated by performing SDS-PAGE, zymogram assay, dynamic light scattering, mass spectrometry, enzyme assays, molecular modeling, circular dichroism spectroscopy, and fluorescence analysis. In addition, cross-linked enzyme aggregates (CLEAs) of EfEstA exhibited high recycling processes and improved stabilities compared to free EfEstA. Furthermore, macroscopic hydrogels of EfEstA could be used as an environment friendly carrier to absorb several metal ions. These characteristics of EfEstA shed light on the design and fabrication of new biomaterials based on bacterial virulence factors for the pharmaceutical, biotechnological, and chemical industries.
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Contents
Abstract ⅰ
List of Figures ⅱ
Ⅰ. Introduction 1
Ⅱ. Materials and methods 3
2.1 Bacterial strains, enzymes, and reagents 3
2.2 Sequence analysis and homology modeling 3
2.3 Cloning, expression, and purification of EfEstA 4
2.4. Enzyme assays 4
2.5. Circular dichroism and fluorescence analysis 5
2.6. Activity and aggregation in high salt conditions 5
2.7. Preparation of crosslinked enzyme aggregates (CLEAs) 6
2.8. Hydrogel formation of EfEstA 6
Ⅲ. Results and Discussion 8
3.1 Bioinformatic analysis of EfEstA 8
3.2 Biochemical analysis of EfEstA 11
3.3 Structural analysis of EfEstA 16
3.4 Salt- and chemical-stabilities of EfEstA 19
3.5 CLEAs of EfEstA 25
3.6 Hydrogel formation of EfEstA 27
Ⅳ. Conclusions 29
Ⅴ. Appendix 47
5.1 Introduction 48
5.2 Materials and Methods 50
5.3 Results and discussion 53
5.4. Conclusions 63
Reference 64
