Substrate-Dependent Alteration in the C- and O-Prenylation Specificities of Cannabis Prenyltransferase

• Published in: Biological & pharmaceutical bulletin 2024/02/17, Vol.47(2), pp.449-453
• Main Authors: Tanaya, Ryosuke, Kodama, Takeshi, Maneenet, Juthamart, Yasuno, Yoko, Nakayama, Atsushi, Shinada, Tetsuro, Takahashi, Hironobu, Ito, Takuya, Morita, Hiroyuki, Awale, Suresh, Taura, Futoshi
• Format: Article
• Language: English
• Subjects: Acids; acylphloroglucinol; antiausterity; Biological activity; biosynthesis; cannabinoid; Cannabis; Catalysts; Isomers; Pancreatic cancer; prenyltransferase; Prenyltransferases; Substrate specificity
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.b23-00868

CsPT4 is an aromatic prenyltransferase that synthesizes cannabigerolic acid (CBGA), the key intermediate of cannabinoid biosynthesis in Cannabis sativa, from olivetolic acid (OA) and geranyl diphosphate (GPP). CsPT4 has a catalytic potential to produce a variety of CBGA analogs via regioselective C-prenylation of aromatic substrates having resorcylic acid skeletons including bibenzyl 2,4-dihydroxy-6-phenylethylbenzoic acid (DPA). In this study, we further investigated the substrate specificity of CsPT4 using phlorocaprophenone (PCP) and 2′,4′,6′-trihydroxydihydrochalcone (THDC), the isomers of OA and DPA, respectively, and demonstrated that CsPT4 catalyzed both C-prenylation and O-prenylation reactions on PCP and THDC that share acylphloroglucinol substructures. Interestingly, the kinetic parameters of CsPT4 for these substrates differed depending on whether they underwent C-prenylation or O-prenylation, suggesting that this enzyme utilized different substrate-binding modes suitable for the respective reactions. Aromatic prenyltransferases that catalyze O-prenylation are rare in the plant kingdom, and CsPT4 was notable for altering the reaction specificity between C- and O-prenylations depending on the skeletons of aromatic substrates. We also demonstrated that enzymatically synthesized geranylated acylphloroglucinols had potent antiausterity activity against PANC-1 human pancreatic cancer cells, with 4′-O-geranyl THDC being the most effective. We suggest that CsPT4 is a valuable catalyst to generate biologically active C- and O-prenylated molecules that could be anticancer lead compounds.