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Language: korean

Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received September 14, 2016
Accepted December 20, 2016

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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대사공학으로 제작된 재조합 Klebsiella pneumoniae를 이용한 아세토인 생산

Acetoin Production Using Metabolically Engineered Klebsiella pneumoniae

장지웅 정휘민 김덕균 오민규^†

Ji-Woong Jang Hwi-Min Jung Duck Gyun Kim Min-Kyu Oh^†

고려대학교 화공생명공학과, 02841 서울특별시 성북구 안암로 145

Department of Chemical and Biological Engineering, Korea University, 145, Anam-ro, Seongbuk-gu, Seoul, 02841, Korea

mkoh@korea.ac.kr

Korean Chemical Engineering Research, April 2017, 55(2), 237-241(5), 10.9713/kcer.2017.55.2.237 Epub 31 March 2017

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Abstract

아세토인(acetoin)은 식품과 화학산업에서 플랫폼 물질로 이용되며 산업적으로 다양한 응용이 가능한 물질이다. 본 연구에서는 대사공학(metabolic engineering)을 이용하여 아세토인의 생산량이 증가한 재조합 Klebsiella pneumoniae를 구축하였다. 우선 2,3-부탄디올(2,3-butanediol)생산을 위해 제작되었던 재조합 K. pneumoniae (KMK-05)에서 두 가지 2,3-butanediol dehydrogenase (budC, dhaD)를 유전체에서 제거하여 아세토인 생산량을 늘리고, 전사인자 중 하나인 AcoK를 제거하여 아세토인을 분해하는 효소의 발현량을 줄였다. 그리고 NADH oxidase를 발현시켜 세포 내 산화 환원 균형(redox balance)을 맞춰 대사흐름을 개선하였다. 이렇게 대사공학을 통해 구축된 재조합 Klebsiella pneumoniae (KJW-03-nox)로 아세토인 생산량과 수율을 높였고, 36시간 동안의 유가식 배양을 진행하여 51 g/L의 아세토인 농도와 최대 생산성 2.6 g/L/h을 달성하였다.

Acetoin is variously applicable platform chemical in chemical and food industry. In this study, Klebsiella pneumoniae was engineered for acetoin production using metabolic engineering. From the recombinant Klebsiella pneumoniae (KMK-05) producing 2,3-butanediol, budC and dhaD genes encoding two 2,3-butanediol dehydrogenases were deleted to reduce 2,3-butanediol production. Furthermore, a transcriptional regulator, AcoK, was deleted to reduce the expression levels of acetoin degrading enzyme. Lastly, NADH oxidase was overexpressed for adjusting intracellular redox balance. The resulting strain (KJW-03-nox) produced considerable amount of acetoin, with concentration reaching 51 g/L with 2.6 g/L/h maximum productivity in 36 h fed-batch fermentation.

Keywords

Metabolic engineering Acetoin 2,3-Butanediol dehydrogenase NADH oxidase

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