Metabolic engineering of hyperthermophilic archaeon thermococcus onnurineus for 3-hydroxypropionate and 3-hydroxyisobutyrate production = 대사공학을 통한 초호열성 고세균 Thermococcus onnurineus 를 이용한 3-hydroxypropionate와 3-hydroxyisobutyrate 생산

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Hyperthermphilic microorganisms has advantages for biochemical production such as reduced risk of bacterial contamination and reduced cooling requirement. Thermococcus onnurineus is a sulfur-reducing hyperthermophilic archaeon which was isolated from a deep-sea hydrothermal vent region. For biochemical production on this beneficial hyperthermophilic microbial platform, T. onnurineus was engineered using transformation with heterogeneous genes. 3-Hydroxypropionate (3-HP) and 3-Hydroxyisobutyrate (3-HIBA) were chosen as model products. For the construction of 3-HP platform, malonyl-CoA reductase (MCR) and malonate semialdehyde reductase (MSR) were introduced on the pyruvate degradation pathway of the host. T. onnurineus has several advantages as biochemical production platform. Cell mass and 3-HP could be produced simultaneously since the optimal temperatures for cell growth and related enzyme activity are close to each other and NAD(P)H, the common cofactor for both reductases, can be sufficiently generated by well-developed redox pathways. A recombinant strain for 3-HP platform was constructed successfully and $140 mg liter^{-1}$ of 3-HP was synthesized in fed-batch fermentation, Continuous fed-batch process increased 3-HP titer up to $191 mg liter^{-1}$ with a productivity was $25.3 mg liter^{-1} h^{-1}$. For the construction of 3-HIBA platform, propionate utilization pathways of the host was engineered with two heterogeneous genes of MCR and 3-Hydroxyisobutyrate dehydrogenase (HIBADH). By fed-batch fermentation, $2.36 mg liter^{-1}$ of 3-HIBA was synthesized. It is the first example of genetically engineered biochemical production platform based on T. onnurineus. However, further study is necessary to increase the productivity and to develop more valuable biochemical compounds.
Advisors
Chang, Yong Keunresearcher장용근researcher
Description
한국과학기술원 :생명화학공학과,
Publisher
한국과학기술원
Issue Date
2017
Identifier
325007
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생명화학공학과, 2017.2,[x, 112 p. :]

Keywords

hyperthermophilic archaeon; T. onnurineus; biochemical production platform; 3-hydroxypropionic acid; 3-hydroxyisobutyric acid; 초고온성 고세균; 서모코커스 온누리니우스; 바이오케미컬 생산 플랫폼; 3-하이드록시프로피온산; 3-하이드록시이소부틸산

URI
http://hdl.handle.net/10203/241875
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=675739&flag=dissertation
Appears in Collection
CBE-Theses_Ph.D.(박사논문)
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