番茄红素是一种具有较高商业价值的萜类化合物,具有良好的抗氧化性,对心血管疾病有预防作用,可通过微生物发酵法高效生产。为了提高酿酒酵母胞内番茄红素积累,过量表达了三酰甘油途径(TAG途径)的关键基因dga1(酰基辅酶A:二酰基甘油酰基转移酶)、pah1(磷脂酸磷酸酶),同时敲入ldp1(脂滴附着蛋白);继续在ypl062w位点敲入来源于肠沙门氏菌的acs1L641P(胞内乙酰CoA合成酶),为番茄红素和脂质载体提供充足的前体;过表达pap1(聚腺苷酸聚合酶)以满足多个基因过表达的转录需求。综上,该文构建了一个适合番茄红素累积的细胞工厂,番茄红素单位产量为109.26 mg/g DCW。该研究通过代谢改造得到1株高产番茄红素重组菌,研究结果为后续进行发酵优化奠定了基础。
Lycopene is a terpenoid compound with high commercial value, which has good antioxidant properties and has a preventive effect on cardiovascular diseases. However, as an antioxidant, lycopene has certain toxicity to cells. According to studies, increasing the intracellular lipid content can effectively alleviate the toxic stress. In this manuscript, we overexpressed the key genes dga1 (acyl-CoA: diacylglycerol acyltransferase) and pah1 (phosphatidic acid phosphatase) of the triacylglycerol pathway (TAG pathway), and knock-in the ldp1 gene (lipid droplet attachment protein) as well, which improved the accumulation of lycopene in the cell. Sufficient precursor is provided by knocking out the ypl062w locus and simultaneously expressing the acs1L641P gene from Salmonella enterica heterologously, for both the lycopene synthesis and the TAG accumulation. pap1 (polynucleotide adenylyltransferase) was overexpressed to meet the transcription requirements of multiple gene overexpression. A cell factory suitable for lycopene accumulation was constructed, in which the unit yield of lycopene reached 109.26 mg/g DCW. A high-yielding lycopene recombinant strain was obtained through metabolic transformation, which laid the foundation for subsequent fermentation optimization.
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