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FCS 및 ECH를 이용한 재조합 대장균에서 L-타이로신으로부터 페놀알데히드 및 멜라닌 색소의 생산
- Author(s)
- 장세영
- Advisor
- 최권영
- Department
- 일반대학원 환경안전공학과
- Publisher
- The Graduate School, Ajou University
- Publication Year
- 2018-02
- Language
- eng
- Keyword
- 멜라닌; protocatechualdehyde
- Abstract
- 본 연구에서는 TAL (tyrosine ammonia-lyase), C3H (p-coumarate 3-hydroxylase), FCS (feruloyl-CoA synthetase) 및 ECH (enoyl-CoA hydratase)를 비롯한 L-타이로신 변환 효소를 발현하도록 대장균을 조작하였다. protocatechualdehyde와 p-hydroxybenzaldehyde 생산 경로로 구성된 이화 회로는 개별 효소의 조합 발현을 통해 재구성되었다. 첫째, FCS와 ECH를 발현하는 세포는 각각 5 mM의 카페익산과 페룰산을 기질로하여 protocatechualdehyde (70.5 %)와 vanillin (96.5 %)로 전환시킬 수 있다. 둘째, TAL과 C3H는 FCS와 ECH와 함께 발현되었다. 이 균주는 L-타이로신을 카페익산으로 전환시킨 다음 protocatechualdehyde로 전환시키는 것을 목표로 하였다. 실험 결과 p-coumarate 3-hydroxylase 과정을 생략하여 생성되는 p-hydroxybenzaldehyde가 protocatechualdehyde와 동시에 생성됨을 알게되었다. 또한, Ascorbic acid는 catechol aldehyde계 멜라닌 생성 억제제로 사용되었으며 protocatechualdehyde와 p-hydroxybenzaldehyde의 생산 수율은 각각 31.0 ± 5.6, 24.0 ± 4.2 mg / L이었다. 마지막으로 카페인 산의 존재 하에서 FCS와 ECH를 동시에 발현시킴으로써 40.9 ± 6.2 mg / L / h의 높은 생산 속도로 카페익산 기반의 멜라닌이 관찰되었다.
- Alternative Abstract
- In this study, we engineered E. coli cells to express L-tyrosine converting enzymes, including tyrosine ammonia-lyase (TAL), p-coumarate 3-hydroxylase (C3H), feruloyl-CoA synthetase (FCS), and enoyl-CoA hydratase/aldolase (ECH). A catabolic circuit, which consisted of the protocatechualdehyde and p-hydroxybenzaldehyde production pathways, was reconstituted through combinatorial expression of discrete enzymes. First, cells expressing FCS and ECH could convert each 5 mM of caffeic acid and ferulic acid into protocatechualdehyde (70.5%) and vanillin (96.5%), respectively. Second, TAL and C3H were co-expressed with FCS and ECH. This strain converted L-tyrosine into caffeic acid, which was then converted into protocatechualdehyde. Ascorbic acid was used as an inhibitor of catechol aldehyde-based melanin formation, and the production yields of protocatechualdehyde and p-hydroxybenzaldehyde were 31.0 ± 5.6 and 24.0 ± 4.2 mg/L, respectively. Finally, caffeic acid-based melanin formation was observed with higher production rate of 40.9 ± 6.2 mg/L/h by co-expressing FCS and ECH in the presence of caffeic acid.
- Fulltext
- Appears in Collections:
- Graduate School of Ajou University > Department of Environmental Safety Engineering > 3. Theses(Master)
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