Enhanced production of individual ganoderic acids by integrating Vitreoscilla haemoglobin expression and calcium ion induction in liquid static cultures of Ganoderma lingzhi

Ganoderic acids produced by Ganoderma exhibit anticancer and antimetastatic activities. A novel approach by combining Vitreoscilla haemoglobin (VHb) expression and calcium ion induction was developed to enhance ganoderic acid (GA) production in liquid static cultures of G. lingzhi. The maximum contents of GA-O, GA-S and GA-Me were 1451.33 +/- 67.50, 1431.23 +/- 79.74 and 1283.81 +/- 85.13 mu g per 100 mg cell weight, respectively under the integrated approach, which are the highest contents as ever reported in Ganoderma. The contents of squalene and lanosterol were increased by 2.0- and 3.0-fold in this case compared with those in the control. The transcription levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase, farnesyl-diphosphate synthase, squalene synthase and cytochrome P450 CYP5150L8 were upregulated by 2.56-, 3.31-, 2.59- and 6.12-fold respectively. Additionally, the expression of VHb improved the ratio of type I to type II GA in liquid static cultivation of G. lingzhi. The transcription levels of cyp512a2, cyp512v2 and cyp512a13, candidate cytochrome P450 genes involved in oxidative modification of the lanostane skeleton in GA biosynthesis, were also increased by 2.28-, 2.65- and 3.54-fold in the VHb-expressing strain respectively. Our results illustrated that the approach described here efficiently improved GA production in G. lingzhi fermentation.