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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Anti-wrinkle Activity of $\beta$-carotene Extracted & Purified from Recombinant Escherichia coli

재조합 대장균으로부터 추출.정제된 베타-카로틴의 주름개선 활성

  • Jo, Ji-Song (Department of Chemical & Biological Engineering and ERI, GyeongSang National University) ;
  • Ku, Bo-Mi (Department of Anatomy & Neurobiology, GyeongSang National University) ;
  • Kang, Sang-Soo (Department of Anatomy & Neurobiology, GyeongSang National University) ;
  • Lee, Jae-Ran (Department of Chemical & Biological Engineering and ERI, GyeongSang National University) ;
  • Kim, You-Geun (KB Cosmetics) ;
  • Lee, He (Department of Chemical & Biological Engineering and ERI, GyeongSang National University) ;
  • Kim, Sung-Bae (Department of Chemical & Biological Engineering and ERI, GyeongSang National University) ;
  • Kim, Seon-Won (Division of Applied Life Science, GyeongSang National University) ;
  • Kim, Chang-Joon (Department of Chemical & Biological Engineering and ERI, GyeongSang National University) ;
  • Chung, In-Young (Department of Electronics and Communications, Kwangwoon University)
  • 조지송 (경상대학교 생명화학공학과 및 공학연구원) ;
  • 구보미 (경상대학교 의과대학 해부학교실) ;
  • 강상수 (경상대학교 의과대학 해부학교실) ;
  • 이재란 (경상대학교 생명화학공학과 및 공학연구원) ;
  • 김유근 (KB코스매틱) ;
  • 이학 (경상대학교 생명화학공학과 및 공학연구원) ;
  • 김성배 (경상대학교 생명화학공학과 및 공학연구원) ;
  • 김선원 (경상대학교 응용생명과학부) ;
  • 김창준 (경상대학교 생명화학공학과 및 공학연구원) ;
  • 정인영 (광운대학교 전자통신공학과)
  • Published : 2008.12.31
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Abstract

This paper described the extraction/purification of $\beta$-carotene from recombinant E.coli and evaluation of anti-wrinkle activity of purified $\beta$-carotene. No significant differences in extraction yields were observed when hexane or isobutyl acetate was used. However, extraction from wet-cell cake resulted in 2-fold higher amount of $\beta$-carotene than that from dry cells. Disruption of 5 g-wet cells by ultrasonic homogenizer, acetone dehydration, extraction with isobutyl acetate resulted in 36 mg of $\beta$-carotene corresponding to 61.2% of recovery. The formation and separation of $\beta$-carotene crystal improved the purity. 633 mg of $\beta$-carotene crystal with 93% purity was obtained from 223 g/L of wet-cell cake harvested from 2.5-L fed-batch culture broth. The cultures of normal human primary fibroblast were performed to investigate the effect of $\beta$-carotene on cytotoxicity as MTT assay and anti-wrinkle activity as collagen synthesis assays. $1.7{\mu}M$ of $\beta$-carotene was found to be optimal concentration at which 1.4-fold higher amount of collagen was synthesized than that in absence of $\beta$-carotene. This indicates that highly purified $\beta$-carotene can be obtained from recombinant E.coli by applying simple method with less toxic solvent and can be used in functional cosmetics as anti-wrinkle agent.

재조합 대장균으로부터 베타-카로틴을 추출/정제하고 정제된 베타-카로틴의 주름개선 성능을 평가하였다. 탈수 용매, 추출 용매에 따른 추출량의 큰 변화는 발견되지 않았으나, wet-cell cake으로부터 추출한 경우가 dry cell로부터 추출한 경우에 비하여 2배 많은 양의 베타-카로틴이 추출되었다. 5 g의 wet-cell cake을 파쇄하고, 아세톤으로 탈수한 후, isobutyl acetate로 추출한 결과 36 mg의 베타-카로틴이 추출되었는데, 이는 61.2% 회수율에 해당한다. 베타-카로틴의 결정을 생성시키고 이를 분리한 결과 순도가 증가하였다. 재조합 대장균 배양액 2.5 L로부터 얻은 wet-cell cake 223 g의 탈수, 세포 파쇄 후 세포 잔사물로부터 베타-카로틴 추출, 농축, 결정화 등을 통하여 최종적으로 순도 93%인 베타-카로틴 결정 633 mg을 얻었다. 사람섬유아세포 배양을 통하여 정제된 베타-카로틴의 세포 독성 및 콜라겐 합성에 미치는 영향을 조사하였다. $1.7{\mu}M$의 베타-카로틴을 첨가 시 세포에 독성을 거의 나타내지 않았으며 콜라겐 합성양은 첨가하지 않은 경우에 비하여 1.4배 증가하였다. 본 결과는 저 유독성 용매를 사용하고 기존보다 간단한 방법을 사용하여 재조합 대장균으로부터 고순도의 베타-카로틴을 정제할 수 있고 이를 기능성 화장품의 주름개선제로 사용될 수 있음을 시사한다.

Keywords

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