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The unicellular green alga  Haematococcus pluvialis  has been recognized as an industry strain to produce simultaneously esterified astaxanthin (EAST) and triacylglycerol (TAG) under stress induction. It is necessary to identify the key enzymes involving in synergistic accumulation of EAST and TAG in  H. pluvialis . In this study, a novel diacylglycerol acyltransferase 1 was systematically characterized by  in vivo  and  in silico  assays. The upregulated expression of  HpDGAT1  gene was positively associated with the significant increase of TAG and EAST contents under stress conditions. Functional complementation by overexpressing  HpDGAT1  in a TAG-deficient yeast strain H1246 revealed that HpDGAT1 could restore TAG biosynthesis and exhibited a high substrate preference for monounsaturated fatty acyl-CoAs (MUFAs) and polyunsaturated fatty acyl-CoAs (PUFAs). Notably, heterogeneous expression of  HpDGAT1  in  Chlamydomonas reinhardtii  and  Arabidopsis thaliana  resulted in a significant enhancement of total oils and concurrently a high accumulation of MUFAs- and PUFAs-rich TAGs. Furthermore, molecular docking analysis indicated that HpDGAT1 contained AST-binding sites. These findings evidence a possible dual-function role for HpDGAT1 involving in TAG and EAST synthesis, demonstrating that it is a potential target gene to enrich AST accumulation in this alga and to design oil production in both commercial algae and oil crops.

frontiers in plant science

Characterization of a Haematococcus pluvialis Diacylglycerol Acyltransferase 1 and Its Potential in Unsaturated Fatty Acid-Rich Triacylglycerol Production