Nomaguchi Tatsuhiro, Maeda Yoshiaki, Liang Yue, Yoshino Tomoko, Asahi Toru, Tanaka Tsuyoshi
Department of Advanced Science and Engineering, Graduate School of Advanced Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan.
J Biosci Bioeng. 2018 Aug;126(2):258-265. doi: 10.1016/j.jbiosc.2018.03.003. Epub 2018 Apr 5.
Microalgal biofuels are desirable alternatives for traditional liquid fuels, but further improvements of productivity are necessary for microalgal biofuels to be economically feasible. As strategies for improving lipid productivity, repression of lipid degradation has a great potential because the lipid accumulation level is determined by a balance between lipid synthesis and degradation. However, studies of lipid degradation and its primary functioning enzyme triacylglycerol (TAG) lipases in microalgae are currently limited. In this study, we report the comprehensive analysis of TAG lipases in the oleaginous diatom Fistulifera solaris JPCC DA0580. First, we confirmed that TAGs in the lipid accumulation organelles, oil bodies, were degraded when the cells were transferred from the nutrient depleted conditions to nutrient replete condition. Further analysis revealed that, after eicosapentaenoic acid (C20:5n-3)-containing TAGs are were degraded, its molecular skeletons were likely to be recycled to produce glycolipids and phospholipids for chloroplast regeneration and cell growth, respectively. Next, we searched putative TAG lipase genes from the draft genome sequence of F. solaris, and discovered 42 candidates based on the amino acid sequence homology analysis. Subsequent transcriptome analysis revealed that 16 of the 42 lipase genes were up-regulated during lipid degradation. Among the up-regulated lipases, a number of enzymes were predicted to localize in endoplasmic reticulum which is closely associated to the lipid accumulation organelles, oil bodies. Our study provided new insights of lipid degradation in oleaginous microalgae, and putative TAG lipases which could be candidates for metabolic engineering in future study to improve microalgal lipid productivity.
微藻生物燃料是传统液体燃料的理想替代品,但要使微藻生物燃料在经济上可行,还需要进一步提高其生产力。作为提高脂质生产力的策略,抑制脂质降解具有很大潜力,因为脂质积累水平由脂质合成与降解之间的平衡决定。然而,目前关于微藻中脂质降解及其主要功能酶三酰甘油(TAG)脂肪酶的研究有限。在本研究中,我们报告了对富油硅藻太阳瘘孔藻JPCC DA0580中TAG脂肪酶的综合分析。首先,我们证实当细胞从营养耗尽条件转移到营养充足条件时,脂质积累细胞器油体中的TAG会被降解。进一步分析表明,含二十碳五烯酸(C20:5n-3)的TAG被降解后,其分子骨架可能分别被循环利用以产生糖脂和磷脂,用于叶绿体再生和细胞生长。接下来,我们从太阳瘘孔藻的基因组草图序列中搜索假定的TAG脂肪酶基因,并基于氨基酸序列同源性分析发现了42个候选基因。随后的转录组分析表明,42个脂肪酶基因中有16个在脂质降解过程中上调。在上调的脂肪酶中,许多酶预计定位于与脂质积累细胞器油体密切相关的内质网中。我们的研究为富油微藻中的脂质降解提供了新见解,并为未来研究中通过代谢工程提高微藻脂质生产力提供了可能的TAG脂肪酶候选基因。