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High-throughput insertional mutagenesis reveals novel targets for enhancing lipid accumulation in Nannochloropsis oceanica
Journal article   Open access   Peer reviewed

High-throughput insertional mutagenesis reveals novel targets for enhancing lipid accumulation in Nannochloropsis oceanica

Christian Sudfeld, Michal Hubacek, Daniel Figueiredo, Mihris I. S. Naduthodi, John van der Oost, Rene H. Wijffels, Maria J. Barbosa and Sarah D'Adamo
Metabolic engineering, Vol.66, pp.239-258
07/01/2021
DOI: 10.1016/j.ymben.2021.04.012
PMID: 33971293
url
https://doi.org/10.1016/j.ymben.2021.04.012View
Published (Version of record) Open Access

Abstract

The microalga Nannochloropsis oceanica is considered a promising platform for the sustainable production of high-value lipids and biofuel feedstocks. However, current lipid yields of N. oceanica are too low for economic feasibility. Gaining fundamental insights into the lipid metabolism of N. oceanica could open up various possibilities for the optimization of this species through genetic engineering. Therefore, the aim of this study was to discover novel genes associated with an elevated neutral lipid content. We constructed an insertional mutagenesis library of N. oceanica, selected high lipid mutants by five rounds of fluorescence-activated cell sorting, and identified disrupted genes using a novel implementation of a rapid genotyping procedure. One particularly promising mutant (HLM23) was disrupted in a putative APETALA2-like transcription factor gene. HLM23 showed a 40%-increased neutral lipid content, increased photosynthetic performance, and no growth impairment. Furthermore, transcriptome analysis revealed an upregulation of genes related to plastidial fatty acid biosynthesis, glycolysis and the Calvin-Benson-Bassham cycle in HLM23. Insights gained in this work can be used in future genetic engineering strategies for increased lipid productivity of Nannochloropsis.
Biotechnology & Applied Microbiology Life Sciences & Biomedicine Science & Technology

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