Enhancement of fatty acid biosynthesis by exogenous acetyl-CoA carboxylase and pantothenate kinase in Escherichia coli
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ORIGINAL RESEARCH PAPER
Enhancement of fatty acid biosynthesis by exogenous acetylCoA carboxylase and pantothenate kinase in Escherichia coli Shusaku Satoh . Miho Ozaki . Saki Matsumoto . Takumi Nabatame . Moena Kaku . Takashi Shudo . Munehiko Asayama . Shigeru Chohnan
Received: 6 May 2020 / Accepted: 1 September 2020 Ó Springer Nature B.V. 2020
Abstract Objectives To establish a technique for efficient fatty acid production through enhancement of coenzyme A (CoA) biosynthesis and malonyl-CoA supply by introducing exogenous pantothenate kinase (coaA) and acetyl-CoA carboxylase (acc) in Escherichia coli. Results The expression of acc, obtained from Corynebacterium glutamicum, accumulated 2.2-fold more fatty acids in E. coli. The addition of coaA from Pseudomonas putaida or fatty acid synthase (fasA) from C. glutamicum resulted in a 3.1- and 3.6-fold increase in fatty acid synthesis in E. coli cells, which expressed acc and coaA, or acc and fasA, respectively. The transformants, simultaneously possessing all three genes, produced 5.6-fold more fatty acids. The strain possessing acc, coaA, and fasA stored 691 mg/L of fatty acids, primarily as phospholipids, inside the inner membrane after 72-h cultivation. In addition,
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02996-w) contains supplementary material, which is available to authorized users. S. Satoh M. Ozaki S. Matsumoto T. Nabatame M. Kaku T. Shudo M. Asayama S. Chohnan (&) Department of Food and Life Sciences, Ibaraki University College of Agriculture, 3-21-1 Chuo, Ami, Ibaraki 300-0393, Japan e-mail: [email protected]
19% of the total CoA pool was occupied by malonylCoA. Conclusions Increased malonyl-CoA significantly contributed to fatty acid production, and the effect was boosted by the expanded total CoA pool. Manipulation of the intracellular CoA species is effective for fatty acid production in E. coli. Keywords Coenzyme A Pantothenate kinase Acetyl-CoA carboxylase Fatty acid synthase Escherichia coli
Introduction Coenzyme A (CoA) is an essential cofactor, along with ATP and NAD(P)H, involved in numerous enzymatic reactions. Hence, biosynthetic pathway engineering and regeneration of these cofactors are achievable to increase useful substance production (Liu and Chen 2011; Wang et al. 2013). Although the techniques to manipulate intracellular ATP and NAD(P)H levels are well studied, there are few reports on CoA cofactor engineering. Furthermore, most of these studies reported the manipulation of malonyl-CoA levels using acetyl-CoA carboxylase (Acc) (Davis et al. 2000; Miyahisa et al. 2005; Leonard et al. 2007; Lu et al. 2008; Zha et al. 2009; Jeon et al. 2011; Meng et al. 2011; Rathnasingh et al.
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Biotechnol Lett
2012; Cheng et al. 2016). Acc catalyzes the carboxylation of acetyl-CoA to form malonyl-CoA. There are structurally two types of bacterial Acc: one consists of four subunits available in most
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