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Metabolic Engineering of Escherichia coli for Enhanced Production of ent-Kaurenoic Acid
- Alternative Title
- Soon Hwan Oh
- Author(s)
- 오순환
- Alternative Author(s)
- Soon Hwan Oh
- Advisor
- 이평천
- Department
- 일반대학원 분자과학기술학과
- Publisher
- The Graduate School, Ajou University
- Publication Year
- 2017-02
- Language
- eng
- Alternative Abstract
- Plant derived terpenoid compounds are used as aromatic and pharmacological agents and they are industrially valuable materials. The ent-kaurenoic acid, a kauranic diterpenoid, and its derivatives have been shown to have a variety of pharmacological activities and the interest in their utilization is increasing. In particular, steviol glycosides derived from ent-kaurenoids are highly sweet and non-caloric natural products, which are valuable in the food and beverage industry. In order to enhance the biosynthetic pathway of ent-kaurenoic acid in E. coli, genome editing techniques were used to insert precursor related genes into the E. coli genomic DNA and induce stable overexpression. The production of ent-kaurenoic acid in the obtained genome edited strain increased 6.3 times compared to the non-overexpressed strain and 1.9 times compared to the overexpressed strain using plasmid system. The gene transcription levels between over-expression systems were compared by quantitative polymerase chain reaction (qPCR) and the transcription level of genome insertion system was lower than that of plasmid system. The ent-kaurene oxidase (KO) wild-type and three mutants were tested in flasks and 1.5 L bioreactor culture. The ent-kaurenoic acid production in the S44T mutant increased 2.2-fold and 1.2-fold in each experiment compared to the wild-type. The results of this study could be used as a basis for the development of microbial biosynthetic strains of other terpenoid compounds as well as ent-kaurenoids and various derivatives.
- Fulltext
- Appears in Collections:
- Graduate School of Ajou University > Department of Molecular Science and Technology > 3. Theses(Master)
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