Laboratory of Cyanobacterial Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Program of Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Sci Rep. 2018 Sep 10;8(1):13510. doi: 10.1038/s41598-018-31789-5.
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.
天然 plsX 和 plsC 基因的过表达首先及时报告了在 Synechocystis sp. PCC 6803 中脂质含量的显著增加。所有过表达(OX)菌株的生长速率、包括叶绿素 a 和类胡萝卜素在内的细胞内色素含量和氧气释放速率与野生型相似。对于脂肪酸组成,饱和脂肪酸,特别是棕榈酸(16:0)在 OX 菌株中明显增加,而不饱和脂肪酸略有增加,特别是亚油酸(18:2)和α-亚麻酸(18:3)。在正常条件下,plsC/plsX 过表达(OX+XC)菌株产生约 24.3%w/dcw 的高脂质含量,并通过乙酸诱导进一步提高到 39.1%w/dcw。该 OX+XC 工程菌株能够降低与聚 3-羟基丁酸(PHB)合成有关的phaA 转录本水平在乙酸处理下。此外,基因转录本的表达水平表明,plsX 和 plsC/plsX 过表达菌株还增加了 accA 转录本的含量,该基因涉及乙酰辅酶 A 的不可逆羧化生成丙二酰辅酶 A。总的来说,与野生型相比,这些过表达菌株通过部分克服脂质生产的限制,显著增加了更高的脂质含量。