Abstract
The overexpression of native plsX and plsC genes involving in fatty acid/phospholipid synthesis first timely-reported the significantly enhanced lipid contents in Synechocystis sp. PCC 6803. Growth rate, intracellular pigment contents including chlorophyll a and carotenoids, and oxygen evolution rate of all overexpressing (OX) strains were normally similar as wild type. For fatty acid compositions, saturated fatty acid, in particular palmitic acid (16:0) was dominantly increased in OX strains whereas slight increases of unsaturated fatty acids were observed, specifically linoleic acid (18:2) and alpha-linolenic acid (18:3). The plsC/plsX-overexpressing (OX + XC) strain produced high lipid content of about 24.3%w/dcw under normal condition and was further enhanced up to 39.1%w/dcw by acetate induction. This OX + XC engineered strain was capable of decreasing phaA transcript level which related to poly-3-hydroxybutyrate (PHB) synthesis under acetate treatment. Moreover, the expression level of gene transcripts revealed that the plsX- and plsC/plsX-overexpression strains had also increased accA transcript amounts which involved in the irreversible carboxylation of acetyl-CoA to malonyl-CoA. Altogether, these overexpressing strains significantly augmented higher lipid contents when compared to wild type by partly overcoming the limitation of lipid production.
Highlights
The third generation of renewable energy resource, especially cyanobacteria, does not compete with human food resources when compared to first and second generations[1]
The enzyme lysophosphatidic acid acyltransferase or 1-sn-glycerol-3-phosphate acyltransferase (LPAAT; PlsC; EC 2.3.2.51) catalyzes the second step in phospholipid biosynthesis, and its function might close proximity to the first step catalyzed by glycerol-3-phosphate acyltransferase (GPAT)
We confirmed the transcription of the introduced genes using Reverse transcription-polymerase chain reaction (RT-PCR) (Fig. 4)
Summary
The third generation of renewable energy resource, especially cyanobacteria, does not compete with human food resources when compared to first and second generations[1]. We constructed three engineered Synechocystis 6803 strains overexpressing plsX-, plsC- and plsC/plsX which significantly enhanced lipid production compared to Synechocystis PCC 6803 wild type.
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