Bornyl Diphosphate Synthase From Cinnamomum burmanni and Its Application for (+)-Borneol Biosynthesis in Yeast

作者：Rui Ma, Ping Su, Juan Guo, Baolong Jin, Qing Ma, Haiyan Zhang, Lingli Chen, Liuying Mao, Mei Tian, Chang‐Jiang‐Sheng Lai, Jinfu Tang, Guanghong Cui, Luqi Huang · 发表于：Frontiers in Bioengineering and Biotechnology · 年份：2021 · DOI：10.3389/fbioe.2021.631863 · 被引用次数：29 · 研究领域：Plant biochemistry and biosynthesis、Natural product bioactivities and synthesis、Pharmacological Effects of Natural Compounds

(+)-Borneol is a desirable monoterpenoid with effective anti-inflammatory and analgesic effects that is known as soft gold. (+)-bornyl diphosphate synthase is the key enzyme in the (+)-borneol biosynthesis pathway. Despite several reported (+)-bornyl diphosphate synthase genes, relatively low (+)-borneol production hinders the attempts to synthesize it using microbial fermentation. Here, we identified the highly specific (+)-bornyl diphosphate synthase CbTPS1 from Cinnamomum burmanni . An in vitro assay showed that (+)-borneol was the main product of CbTPS1 (88.70% of the total products), and the K m value was 5.11 ± 1.70 μM with a k cat value of 0.01 s –1 . Further, we reconstituted the (+)-borneol biosynthetic pathway in Saccharomyces cerevisiae . After tailored truncation and adding Kozak sequences, the (+)-borneol yield was improved by 96.33-fold to 2.89 mg⋅L –1 compared with the initial strain in shake flasks. This work is the first reported attempt to produce (+)-borneol by microbial fermentation. It lays a foundation for further pathway reconstruction and metabolic engineering production of this valuable natural monoterpenoid.