title: Identification of a Chlamydomonas plastidial 2-lysophosphatidic acid acyltransferase and its use to engineer microalgae with increased oil content.
abstract: Despite a strong interest in microalgal oil production, our understanding of the biosynthetic pathways that produce algal lipids and the genes involved in the biosynthetic processes remains incomplete. Here, we report that Chlamydomonas reinhardtii Cre09.g398289 encodes a plastid-targeted 2-lysophosphatidic acid acyltransferase (CrLPAAT1) that acylates the sn-2 position of a 2-lysophosphatidic acid to form phosphatidic acid, the first common precursor of membrane and storage lipids. In vitro enzyme assays showed that CrLPAAT1 prefers 16:0-CoA to 18:1-CoA as an acyl donor. Fluorescent protein-tagged CrLPAAT1 was localized to the plastid membrane in C. reinhardtii cells. Furthermore, expression of CrLPAAT1 in plastids led to a > 20% increase in oil content under nitrogen-deficient conditions. Taken together, these results demonstrate that CrLPAAT1 is an authentic plastid-targeted LPAAT in C. reinhardtii, and that it may be used as a molecular tool to genetically increase oil content in microalgae.
authors.affiliation: Department of Life Science, Pohang University of Science and Technology, Pohang, Korea.
authors.forename: Yasuyo
authors.initials: Y
authors.lastname: Yamaoka
authors.affiliation: Institut de Biosciences et Biotechnologies, CEA Cadarache, Saint-Paul-lez-Durance, France.
authors.forename: Dorine
authors.initials: D
authors.lastname: Achard
authors.affiliation: Department of Life Science, Pohang University of Science and Technology, Pohang, Korea.
authors.forename: Sunghoon
authors.initials: S
authors.lastname: Jang
authors.affiliation: Institut de Biosciences et Biotechnologies, CEA Cadarache, Saint-Paul-lez-Durance, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Legéret
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Sakura-Ku, Saitama, Japan.
authors.forename: Shogo
authors.initials: S
authors.lastname: Kamisuki
authors.affiliation: Department of Life Science, Pohang University of Science and Technology, Pohang, Korea.
authors.forename: Donghwi
authors.initials: D
authors.lastname: Ko
authors.affiliation: Institut de Biosciences et Biotechnologies, CEA Cadarache, Saint-Paul-lez-Durance, France.
authors.forename: Miriam
authors.initials: M
authors.lastname: Schulz-Raffelt
authors.affiliation: Department of Life Science, Pohang University of Science and Technology, Pohang, Korea.
authors.forename: Yeongho
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Life Science, Pohang University of Science and Technology, Pohang, Korea.
authors.forename: Won-Yong
authors.initials: WY
authors.lastname: Song
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Sakura-Ku, Saitama, Japan.|JST, CREST, Chiyoda-ku, Tokyo, Japan.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Nishida
authors.affiliation: Institut de Biosciences et Biotechnologies, CEA Cadarache, Saint-Paul-lez-Durance, France. yonghua.li@cea.fr.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Li-Beisson
authors.affiliation: Department of Life Science, Pohang University of Science and Technology, Pohang, Korea. ylee@postech.ac.kr.|Department of Integrative Bioscience & Biotechnology, Pohang University of Science and Technology, Pohang, Korea. ylee@postech.ac.kr.
authors.forename: Youngsook
authors.initials: Y
authors.lastname: Lee
mesh_terms: D000217:Acyltransferases / Q000235:genetics*; D002696:Chlamydomonas / Q000201:enzymology*; D058086:Microalgae / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D010938:Plant Oils / Q000378:metabolism; D018087:Plastids / Q000201:enzymology*
chemical_list: D010938:Plant Oils; D000217:Acyltransferases; C018764:2-acylglycerophosphate acyltransferase
keywords: acyl specificity; lysophosphatidic acid acyltransferase; microalgae; oil content; plastid transformation; triacylglycerols