Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Jiangsu Provincial Key Lab for the Chemistry and Utilization of Agro-Forest Biomass, College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, PR China.
J Agric Food Chem. 2024 Aug 28;72(34):18890-18897. doi: 10.1021/acs.jafc.4c05387. Epub 2024 Aug 14.
Bark beetles, major pests that bore into forest stems, cause significant economic damage to forests globally. (+)-α-Pinene is the precursor to (+)--verbenol, a crucial component of the aggregation pheromones produced by bark beetles. This paper describes the synthesis of (+)--verbenol in . Initially, the truncation position of (+)-α-pinene synthase (PS30 from ) and monoterpene precursor (geranyl diphosphate/neryl diphosphate) synthases were evaluated. Neryl diphosphate synthase from (NPPS1) and truncated (+)-α-pinene synthase (PS30-39) were selected as promising candidates. Subsequently, the titer of (+)-α-pinene was significantly increased 8.9-fold by using the fusion tag CM29, which enhanced the solubility of PS30-39. In addition, by optimizing expression elements (ribosomal binding sites, linkers, and up elements) and overexpressing CM29*PS30-39, a yield of 134.12 mg/L (+)-α-pinene was achieved. Finally, the first synthesis of enantiopure (+)--verbenol was achieved by introducing a cytochrome P450 mutant from (P450), resulting in a yield of 11.13 mg/L. This study lays the groundwork for developing verbenol-based trapping technology for controlling bark beetles.
松小蠹是钻入森林树干的主要害虫，给全球森林造成了重大的经济损失。(+)-α-蒎烯是(+)-马鞭草烯醇的前体，马鞭草烯醇是松小蠹聚集信息素的重要组成部分。本文描述了 (+)-马鞭草烯醇在 的合成。首先，评估了(+)α-蒎烯合酶（来自 的 PS30）和单萜前体（香叶基二磷酸/橙花基二磷酸）合酶的截断位置。选择来自 的橙花基二磷酸合酶（NPPS1）和截断的(+)-α-蒎烯合酶（PS30-39）作为有前途的候选物。随后，使用融合标签 CM29 可将 (+)-α-蒎烯的产量显著提高 8.9 倍，这增强了 PS30-39 的可溶性。此外，通过优化表达元件（核糖体结合位点、接头和上游元件）并过表达 CM29*PS30-39，实现了 134.12mg/L 的(+)-α-蒎烯产量。最后，通过引入来自 的细胞色素 P450 突变体（P450）实现了对映纯(+)-马鞭草烯醇的首次 合成，产量为 11.13mg/L。这项研究为开发基于马鞭草烯醇的诱捕技术来控制松小蠹奠定了基础。
