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  • TAG pathway engineering via GPAT2 concurrently potentiates abiotic stress tolerance and oleaginicity in Phaeodactylum tricornutum.

TAG pathway engineering via GPAT2 concurrently potentiates abiotic stress tolerance and oleaginicity in Phaeodactylum tricornutum.

Biotechnology for biofuels (2020-09-19)
Xiang Wang, Si-Fen Liu, Ruo-Yu Li, Wei-Dong Yang, Jie-Sheng Liu, Carol Sze Ki Lin, Srinivasan Balamurugan, Hong-Ye Li
ABSTRACT

Despite the great potential of marine diatoms in biofuel sector, commercially viable biofuel production from native diatom strain is impractical. Targeted engineering of TAG pathway represents a promising approach; however, recruitment of potential candidate has been regarded as critical. Here, we identified a glycerol-3-phosphate acyltransferase 2 (GPAT2) isoform and overexpressed in Phaeodactylum tricornutum. GPAT2 overexpression did not impair growth and photosynthesis. GPAT2 overexpression reduced carbohydrates and protein content, however, lipid content were significantly increased. Specifically, TAG content was notably increased by 2.9-fold than phospho- and glyco-lipids. GPAT2 overexpression elicited the push-and-pull strategy by increasing the abundance of substrates for the subsequent metabolic enzymes, thereby increased the expression of LPAAT and DGAT. Besides, GPAT2-mediated lipid overproduction coordinated the expression of NADPH biosynthetic genes. GPAT2 altered the fatty acid profile in TAGs with C16:0 as the predominant fatty acid moieties. We further investigated the impact of GPAT2 on conferring abiotic stress, which exhibited enhanced tolerance to hyposaline (70%) and chilling (10 ºC) conditions via altered fatty acid saturation level. Collectively, our results exemplified the critical role of GPAT2 in hyperaccumulating TAGs with altered fatty acid profile, which in turn uphold resistance to abiotic stress conditions.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2′,7′-Dichlorofluorescin diacetate, ≥97%
Sigma-Aldrich
Triton X-100, laboratory grade