Enhancing lipid productivity in Nannochloropsis salina by overexpression of endogenous glycerol-3-phosphate dehydrogenase

Koh, Hyun Gi; Chang, Yong Keun; Kang, Nam Kyu

doi:10.1007/s10811-023-03141-6

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“…Typically, the overexpression of genes involved in fatty acid synthesis has been mainly conducted, such as acetyl-CoA carboxylase (ACCase), 3-ketoacyl-ACP synthase (KAS), fatty acid synthase (FAS), and pyruvate dehydrogenase kinase (PDK) (Kang et al, 2022). Metabolic engineering of the TAG synthesis pathway has also been explored by regulating the key genes, including glycerol-3-phosphate dehydrogenase (GPDH), glycerol-3phosphate acyltransferases (GPAT), lysophosphatidic acid acyltransferases (LPAAT), phosphatidate phosphatase (PAP), and diacylglycerol transferase (DGAT) (Kang et al, 2022;Koh et al, 2023). Enhancing the Calvin cycle, gluconeogenesis, and glycolysis have also been studied to increase carbon flux towards lipids (Koh et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Increasing lipid production in Chlamydomonas reinhardtii through genetic introduction for the overexpression of glyceraldehyde-3-phosphate dehydrogenase

Shin,

Koh,

Park

et al. 2024

Front. Bioeng. Biotechnol.

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Microalgae, valued for their sustainability and CO2 fixation capabilities, are emerging as promising sources of biofuels and high-value compounds. This study aimed to boost lipid production in C. reinhardtii by overexpressing chloroplast glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a key enzyme in the Calvin cycle and glycolysis, under the control of a nitrogen-inducible NIT1 promoter, to positively impact overall carbon metabolism. The standout transformant, PNG#7, exhibited significantly increased lipid production under nitrogen starvation, with biomass rising by 44% and 76% on days 4 and 16, respectively. Fatty acid methyl ester (FAME) content in PNG#7 surged by 2.4-fold and 2.1-fold, notably surpassing the wild type (WT) in lipid productivity by 3.4 and 3.7 times on days 4 and 16, respectively. Transcriptome analysis revealed a tenfold increase in transgenic GAPDH expression and significant upregulation of genes involved in fatty acid and triacylglycerol synthesis, especially the gene encoding acyl-carrier protein gene (ACP, Cre13. g577100. t1.2). In contrast, genes related to cellulose synthesis were downregulated. Single Nucleotide Polymorphism (SNP)/Indel analysis indicated substantial DNA modifications, which likely contributed to the observed extensive transcriptomic and phenotypic changes. These findings suggest that overexpressing chloroplast GAPDH, coupled with genetic modifications, effectively enhances lipid synthesis in C. reinhardtii. This study not only underscores the potential of chloroplast GAPDH overexpression in microalgal lipid synthesis but also highlights the expansive potential of metabolic engineering in microalgae for biofuel production.

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Section: Introductionmentioning

confidence: 99%