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Efficient synthesis of functional isoprenoids from acetoacetate through metabolic pathway-engineered Escherichia coli

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Abstract

We show here an efficient synthesis system of isoprenoids from acetoacetate as the main substrate. We expressed in Escherichia coli a Streptomyces mevalonate pathway gene cluster starting from HMG-CoA synthase and including isopentenyl diphosphate isomerase (idi) type 2 gene and the yeast idi type 1 and rat acetoacetate-CoA ligase (Aacl) genes. When the α-humulene synthase (ZSS1) gene of shampoo ginger was expressed in this transformant, the resultant E. coli produced 958 μg/mL culture of α-humulene with a lithium acetoacetate (LAA) supplement, which was a 13.6-fold increase compared with a control E. coli strain expressing only ZSS1. Next, we investigated if this E. coli strain engineered to utilize acetoacetate can synthesize carotenoids effectively. When the crtE, crtB, and crtI genes required for lycopene synthesis were expressed in the transformant, lycopene amounts reached 12.5 mg/g dry cell weight with addition of LAA, an 11.8-fold increase compared with a control expressing only the three crt genes. As for astaxanthin production with the E. coli transformant, in which the crtE, crtB, crtI, crtY, crtZ, and crtW genes were expressed, the total amount of carotenoids produced (astaxanthin, lycopene, and phytoene) was significantly increased to 7.5 times that of a control expressing only the six crt genes.

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Acknowledgements

This work has been supported as a part of the Research and Development Program for New Bio-Industry Initiatives (2006–2010) of Bio-Oriented Technology Research Advancement Institution (BRAIN).

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Authors and Affiliations

  1. Central Laboratories for Frontier Technology, Kirin Holdings Co., Ltd., i-BIRD 3-570, Suematsu, Nonoichi-machi, Ishikawa, 921-8836, Japan

    Hisashi Harada & Norihiko Misawa

  2. Department of Bioscience, Graduate School of Agriculture, Kinki University, Nakamachi, Nara, 631-8505, Japan

    Fengnian Yu, Sho Okamoto & Ryutaro Utsumi

  3. Laboratory of Cell Biotechnology, Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

    Tomohisa Kuzuyama

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  1. Hisashi Harada
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  2. Fengnian Yu
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  3. Sho Okamoto
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  4. Tomohisa Kuzuyama
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  5. Ryutaro Utsumi
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Correspondence to Norihiko Misawa.

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Harada, H., Yu, F., Okamoto, S. et al. Efficient synthesis of functional isoprenoids from acetoacetate through metabolic pathway-engineered Escherichia coli . Appl Microbiol Biotechnol 81, 915–925 (2009). https://doi.org/10.1007/s00253-008-1724-7

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