Department of Biological Chemistry, Chubu University, 1200 Matsumoto-cho, Kasugai, 487-8501 Japan.
Japan Science and Technology Agency, CREST, 4-1-8 Honmachi, Kwaguchi, Saitama 332-0012, Japan.
Plant Cell Physiol. 2022 Jan 25;63(1):82-91. doi: 10.1093/pcp/pcab147.
Cyanobacterial mutants defective in acyl-acyl carrier protein synthetase (Aas) produce free fatty acids (FFAs) because the FFAs generated by deacylation of membrane lipids cannot be recycled. An engineered Aas-deficient mutant of Synechocystis sp. PCC 6803 grew normally under low-light (LL) conditions (50 µmol photons m-2 s-1) but was unable to sustain growth under high-light (HL) conditions (400 µmol photons m-2 s-1), revealing a crucial role of Aas in survival under the HL conditions. Several-times larger amounts of FFAs were produced by HL-exposed cultures than LL-grown cultures. Palmitic acid accounted for ∼85% of total FFAs in HL-exposed cultures, while C18 fatty acids (FAs) constituted ∼80% of the FFAs in LL-grown cultures. Since C16 FAs are esterified to the sn-2 position of lipids in the Synechocystis species, it was deduced that HL irradiation activated deacylation of lipids at the sn-2 position. Heterologous expression of FarB, the FFA exporter protein of Neisseria lactamica, prevented intracellular FFA accumulation and rescued the growth defect of the mutant under HL, indicating that intracellular FFA was the cause of growth inhibition. FarB expression also decreased the 'per-cell' yield of FFA under HL by 90% and decreased the proportion of palmitic acid to ∼15% of total FFA. These results indicated that the HL-induced lipid deacylation is triggered not by strong light per se but by HL-induced damage to the cells. It was deduced that there is a positive feedback loop between HL-induced damage and lipid deacylation, which is lethal unless FFA accumulation is prevented by Aas.
产氰蓝细菌突变体中酰基辅酶 A 合成酶(Aas)缺陷,因为膜脂去酰化产生的游离脂肪酸(FFA)不能被回收。Synechocystis sp. PCC 6803 的工程 Aas 缺陷突变体在低光(LL)条件(50 µmol 光子 m-2 s-1)下正常生长,但在高光(HL)条件(400 µmol 光子 m-2 s-1)下无法维持生长,表明 Aas 在 HL 条件下的生存中起着至关重要的作用。与 LL 生长培养物相比,HL 暴露培养物产生的 FFA 量要大几倍。HL 暴露培养物中棕榈酸占总 FFA 的约 85%,而 LL 生长培养物中 C18 脂肪酸(FA)占 FFA 的约 80%。由于 C16 FAs 酯化到 Synechocystis 物种的 sn-2 位置的脂质中,因此推断 HL 照射激活了 sn-2 位置的脂质去酰化。Neisseria lactamica 的 FFA 外排蛋白 FarB 的异源表达防止了细胞内 FFA 的积累,并挽救了突变体在 HL 下的生长缺陷,表明细胞内 FFA 是生长抑制的原因。FarB 表达还使 HL 下“每细胞”FFA 产量降低了 90%,并将棕榈酸的比例降低到总 FFA 的约 15%。这些结果表明,HL 诱导的脂质去酰化不是由强光本身引起的,而是由 HL 诱导的细胞损伤引起的。推断 HL 诱导的损伤和脂质去酰化之间存在正反馈回路,如果不防止 Aas 积累 FFA,该回路是致命的。