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Overexpression of acetyl-CoA carboxylase increases fatty acid production in the green alga Chlamydomonas reinhardtii

  • Duo Chen
  • Xue Yuan
  • Limin Liang
  • Kui Liu
  • Haoying Ye
  • Zhiyu Liu
  • Yanfei Liu
  • Luqiang Huang
  • Wenjin He
  • Youqiang Chen
  • Yanding ZhangEmail author
  • Ting XueEmail author
Original Research Paper
  • 43 Downloads

Abstract

Chlamydomonas reinhardtii is a photosynthetic unicellular model algae with multiple biotechnological advantages, and its fatty acids can be used to produce biofuels. Numerous studies suggest that acetyl-coA carboxylase (ACCa) catalyzes the first committed and rate-limiting step of fatty acid biosynthesis, thereby playing a central role in oil accumulation. Here, we cloned and overexpressed ACCa in C. reinhardtii to directly evaluate its effect on fatty acid synthesis. GC–MS analysis found that the unsaturated FAs contents of the CW15-24 and CW15-85 strains were 55.45% and 56.15%, which were significantly enriched compared to the wild type CW15 (48.39%). Under the optimized conditions, the content of lipid by overexpressed the ACCa gene in the mutant CW15-85 (0.46 g/l) was 1.16-fold greater than control through optimization of N and P sources. Altogether, our data clearly demonstrate that ACCa overexpression in C. reinhardtii can directly increase the synthesis of fatty acids.

Keywords

Acetyl-CoA carboxylase Biofuel Chlamydomonas reinhardtii Fatty acids Lipid 

Notes

Acknowledgements

We would like to thank the native English speaking scientists of Elixigen Company (Huntington Beach, California) for editing our manuscript. This work was supported by the Natural Science Foundation of Fujian Province, China (Grant Number 2017J01622) and the Sugar Crop Research System (Grant Number CARS-170501).

Supporting information

Supplementary Fig. 1—Characterization of the PHKA overexpression construct by PCR detection of Acca and mCherry and restriction analysis. (A) Acca amplification from three independent PHKA clones confirms insertion of Acca coding sequence (lane M, molecular weight marker; lanes 1-3, Acca). (B) Restriction validation of PHKA clones (lane M, molecular weight marker; lane 1, products of PHKA; lanes 2-3, EcoR I digestion; lanes 4-5, EcoR V and EcoR I digestion). (C) mCherry amplification from C. reinhardtii mutant genomic DNA (lane M, molecular weight marker; lanes 1-5, mCherry).

Supplementary Fig. 2−Protein spot hybridization using anti-mCherry and anti-GAPDH antibodies. (A) GAPDH (endogenous control) is detected in wild type (CW15) and ACCa-overexpressing transgenic clones (CW15-24, CW15-85). (B) mCherry is detected only in the ACCa-overexpressing (CW15-24, CW15-85) clones, suggesting successful transfection of the PHKA overexpression vector.

Supplementary material

10529_2019_2715_MOESM1_ESM.docx (268 kb)
Supplementary material 1 (DOCX 268 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.The Public Service Platform for Industrialization Development Technology of Marine Biological Medicine and Product of State Oceanic Administration, Key Laboratory of Developmental and Neural Biology, College of Life SciencesFujian Normal UniversityFuzhouPeople’s Republic of China
  2. 2.Center of Engineering Technology Research for Microalgae Germplasm Improvement of Fujian, Southern Institute of OceanographyFujian Normal UniversityFuzhouPeople’s Republic of China
  3. 3.Fisheries Research Institute of FujianXiamenPeople’s Republic of China
  4. 4.Fujian Fisheries Technical Extension CenterFuzhouPeople’s Republic of China

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