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Title 

CRISPR interference-guided balancing of a biosynthetic mevalonate pathway increases terpenoid production

 

크리스퍼 간섭을 이용한 메발로네이트 대사경로의 조절을 통해 터펜 생산 증대

Authors 

Seong Keun KimGui Hwan HanWonjae SeongHaseong KimS W KimDae-Hee LeeSeung Goo Lee

Publisher 

Elsevier

Issue Date 

2016

Citation 

Metabolic Engineering, vol. 38, no. 0, pp. 228-240

Keywords 

CRISPR interferenceEscherichia coliMevalonate pathwayTerpenoid

Abstract 

Methods for simple and efficient regulation of metabolic pathway genes are essential for maximizing product titers and conversion yields, and for minimizing the metabolic burden caused by heterologous expression of multiple genes often in the operon context. Clustered regularly interspaced short palindromic repeats (CRISPR) interference (CRISPRi) is emerging as a promising tool for transcriptional modulation. In this study, we developed a regulatable CRISPRi system for fine-tuning biosynthetic pathways and thus directing carbon flux toward target product synthesis. By exploiting engineered Escherichia coli harboring a biosynthetic mevalonate (MVA) pathway and plant-derived terpenoid synthases, the CRISPRi system successfully modulated the expression of all the MVA pathway genes in the context of operon and blocked the transcription of the acetoacetyl-CoA thiolase enzyme that catalyzes the first step in the MVA pathway. This CRISPRi-guided balancing of expression of MVA pathway genes led to enhanced production of (-)-α-bisabolol (C15) and lycopene (C40) and alleviation of cell growth inhibition that may be caused by expression of multiple enzymes or production of toxic intermediate metabolites in the MVA pathway. Coupling CRISPRi to cell growth by regulating an endogenous essential gene (ispA) increased isoprene (C5) production. The regulatable CRISPRi system proved to be a robust platform for systematic modulation of biosynthetic and endogenous gene expression, and can be used to tune biosynthetic metabolic pathways. Its application can enable the development of microbial ‘smart cell’ factories that can produce other industrially valuable products in the future.

ISSN 

1096-7176

Link 

http://dx.doi.org/10.1016/j.ymben.2016.08.006

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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