Biosynthesis of a Novel Ganoderic Acid in Saccharomyces cerevisiae and Research of its Antitumor Activity

Ganoderic acids (GAs), mainly from the traditional Chinese medicinal Ganoderma lucidum, possess antitumor, anti-metastasis and other significant pharmacological activities. Due to the difficulty to purify the GAs monomer with low yield and similar structure, the pharmacological activities of most of GA have not been clarified, which hindered their widespread application. In this study, we expressed our previously identified two CYPs, CYP5150L8 and CYP5139G1 in Saccharomyces cerevisiae to generate two plasmid-containing yeast strain Y2P, which can produce 3,28-dihydroxy-lanosta-8,24-dien-26-oic acid (DHLDOA) at concentration of 0.6 mg/L. To improve the production of DHLDOA, we transfer CYP partner, cytochrome P450 reductase, into the CYP5150 and CYP5139G1 co-expression Y2P strain to obtain 3 plasmids co-transformation strain Y3P, and used 3 antibiotics to avoid plasmid loss during cell cultivation. The DHLDOA production of S. cerevisiae Y3P reached to 2.9 and 11.1 mg/L without or with adding hygromycin, G418, and zeocin, respectively. In addition, it was found that DHLDOA was effective against non-small cell lung cancer but had low toxicity to normal cells. This study may facilitate the wide-spread application of GA-DHLDOA and further improvement of DHLDOA production by the yeast fermentation.