Efficient synthesis of functional isoprenoids from acetoacetate through metabolic pathway-engineered <Emphasis Type="Italic">Escherichia coli</Emphasis> |
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Authors: | Hisashi?Harada Fengnian?Yu Sho?Okamoto Tomohisa?Kuzuyama Ryutaro?Utsumi Email author" target="_blank">Norihiko?MisawaEmail author |
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Institution: | (1) Central Laboratories for Frontier Technology, Kirin Holdings Co., Ltd., i-BIRD 3-570, Suematsu, Nonoichi-machi Ishikawa, 921-8836, Japan;(2) Department of Bioscience, Graduate School of Agriculture, Kinki University, Nakamachi Nara, 631-8505, Japan;(3) Laboratory of Cell Biotechnology, Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan |
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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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Keywords: | α -Humulene Lycopene Astaxanthin Sesquiterpene Carotenoids |
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