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