Effect of Temperature-Shift and Temperature-Constant Cultivation on the Monacolin K Biosynthetic Gene Cluster Expression in sp.

In this study, the effects of temperature-shift (from 30 to 25 °C) and temperature-constant (at 30 °C) cultivation on the mass of CG-6 mycelia and concentration of the produced monacolin K (MK) were monitored. The expression levels of the MK biosynthetic genes of CG-6 at constant and variable culture temperatures were analysed by real-time quantitative polymerase chain reaction (RT-qPCR). The total protein was collected and determined by liquid chromatography-electrospray ionisation with tandem mass spectrometry (LC-ESI-MS/MS). Results showed that the maximum mycelial mass in temperature-shift cultivation was only 0.477 g of dry cell mass per dish, which was lower than that in temperature-constant cultivation (0.581 g of dry cell mass per dish); however, the maximum concentration of MK in temperature-shift cultivation (34.5 µg/mL) was 16 times higher than that in temperature-constant cultivation at 30 °C (2.11 µg/mL). Gene expression analysis showed that the expression of the MK biosynthetic gene cluster at culture temperature of 25 °C was higher than that at 30 °C, which was similar to the trend of the MK concentration, except for individual and genes. Analysis of differential protein expression revealed that 2016 proteins were detected by LC-ESI-MS/MS. The expression level of efflux pump protein coded by the gene exhibited the same upregulated trend as the expression of in temperature-shift cultivation. Temperature-shift cultivation enhanced the expression of proteins in the secondary metabolite production pathway, but suppressed the expression of proteins involved in the mycelial growth.