Department of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, Denmark.
Mosspiration Biotech, Hørsholm, Denmark.
PLoS One. 2020 Dec 7;15(12):e0243620. doi: 10.1371/journal.pone.0243620. eCollection 2020.
Plant-derived terpenoids are extensively used in perfume, food, cosmetic and pharmaceutical industries, and several attempts are being made to produce terpenes in heterologous hosts. Native hosts have evolved to accumulate large quantities of terpenes in specialized cells. However, heterologous cells lack the capacity needed to produce and store high amounts of non-native terpenes, leading to reduced growth and loss of volatile terpenes by evaporation. Here, we describe how to direct the sesquiterpene patchoulol production into cytoplasmic lipid droplets (LDs) in Physcomitrium patens (syn. Physcomitrella patens), by attaching patchoulol synthase (PTS) to proteins linked to plant LD biogenesis. Three different LD-proteins: Oleosin (PpOLE1), Lipid Droplet Associated Protein (AtLDAP1) and Seipin (PpSeipin325) were tested as anchors. Ectopic expression of PTS increased the number and size of LDs, implying an unknown mechanism between heterologous terpene production and LD biogenesis. The expression of PTS physically linked to Seipin increased the LD size and the retention of patchoulol in the cell. Overall, the expression of PTS was lower in the anchored mutants than in the control, but when normalized to the expression the production of patchoulol was higher in the seipin-linked mutants.
植物源萜类化合物广泛应用于香水、食品、化妆品和制药行业，目前正在尝试在异源宿主中生产萜类化合物。天然宿主已经进化到在特化细胞中积累大量萜类化合物。然而，异源细胞缺乏产生和储存大量非天然萜类化合物的能力，导致生长减少和挥发性萜类化合物通过蒸发而损失。在这里，我们描述了如何通过将贴梗醇合酶 (PTS) 连接到与植物脂滴生物发生相关的蛋白质上，将倍半萜贴梗醇定向生产到Physcomitrium patens（同物异名Physcomitrella patens）的细胞质脂滴 (LD) 中。我们测试了三种不同的 LD 蛋白：油蛋白 (PpOLE1)、脂滴相关蛋白 (AtLDAP1) 和 Seipin (PpSeipin325) 作为锚定蛋白。PTS 的异位表达增加了 LD 的数量和大小，这暗示了异源萜类化合物生产和 LD 生物发生之间存在未知的机制。PTS 与 Seipin 物理连接的表达增加了 LD 的大小和细胞内贴梗醇的保留。总的来说，锚定突变体中 PTS 的表达低于对照，但归一化到表达量后，与 seipin 相关的突变体中贴梗醇的产量更高。
