Earticle

초록

영어

Using Bacillus subtilis spore display system, with cotG as an anchoring motif, levansucrase from Zymomonas mobilis, was displayed on the outer surface of Bacillus subtilis spore. Flow cytometry of DB104 (pSDJH-cotG-levU) spore, proved the surface localization of CotG-LevU fusion protein on the spore compared to that of DB104. Enzymatic activity of DB104 (pSDJH-cotG-levU) spore showed more than 1.5 times higher levansucrase specific activity compared to that of the host spore, which is a remarkable increase of enzymatic activity considering the existence of sacA (sucrase) and sacB (levansucrase) in the Bacillus subtilis chromosome. The spore integrity, revealed by sporulation frequency test after heat and lysozyme treatment of spore, did not changed at all in spite of the CotG-LevU fusion protein incorporation into the spore coat layer during spore formation process. These data prove again that Bacillus subtilis spore could be considered as good live immobilization vehicle for efficient bioconversion process.

목차

Abstract
 1. 서론
 2. 재료 및 방법
  2.1. 균주 및 배양 조건
  2.2. 유전자 증폭 (PCR amplification)
  2.3. 유전자 조작 (DNA manipulation)
  2.4. 고초균으로의 형질 전환 (Transformation into Bacillus subtilis)
  2.5. 고초균 포자 분리 및 정제 방법 (Purification of Bacillus subtilis spore)
  2.6. 유세포 분석 (Flow cytometric analysis)
  2.7. levansucrase 효소 역가 측정
  2.8. 포자 형성 빈도 측정 (Sporulation frequency measurement)
 3. 결과 및 고찰
  3.1. CotG-LevU 융합 단백질 발현 벡터 제작 및 고초균 균주의 구축
  3.2. 유세포 분석기를 이용한 levansucrase의 포자 표면 발현
  3.3. 포자 표면 발현된 levansucrase 효소 활성 측정
  3.4. 열처리와 lysozyme 처리를 통한 포자 안정성 검증
 4. 결론
 감사
 References

키워드

저자

• 김준형 [ June-Hyung Kim | 동아대학교 화학공학과 ]
• 최수근 [ Soo-Keun Choi | 한국생명공학연구원 ]
• 정흥채 [ Heung-Chae Jung | 한국생명공학연구원 ]
• 반재구 [ Jae-Gu Pan | 한국생명공학연구원 ]
• 김병기 [ Byung-Gee Kim | 서울대학교 화학생물공학부 ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  KSBB Journal

• 간기

  격월간

• pISSN

  1225-7117

• 수록기간

  2009~2013

• 십진분류

  KDC 476 DDC 576