Construction of an Efficient and Robust Aspergillus terreus Cell Factory for Monacolin J Production

Abstract

Monacolin J is a key precursor for the synthesis of the cholesterol-lowering drug simvastatin. Industrially, monacolin J is manufactured through the alkaline hydrolysis of the fungal polyketide lovastatin, which is relatively complex and environmentally unfriendly. A cell factory for monacolin J production was created by heterologously introducing lovastatin hydrolase into Aspergillus terreus in our previous study. However, residual lovastatin remained a problem for the downstream product purification. In this study, we used combined metabolic engineering strategies to create a more efficient and robust monacolin J-producing cell factory that completely lacks lovastatin residue. The complete deletion of the key gene lovF blocked the biosynthesis of lovastatin and led to a large accumulation of monacolin J without any lovastatin residue. Additionally, the overexpression of the specific transcription factor lovE under the PgpdAt promoter further increased the titer of monacolin J by 52.5% to 5.5 g L–1. Interestingly, the fermentation robustness was also significantly improved by the expression of lovE. This improvement not only avoids the process of alkaline hydrolysis but also simplifies the downstream separation process.

Growing a circular economy with fungal biotechnology: a white paper

“…Current research is focused on re-routing lovastatin biosynthesis to one of its biosynthetic intermediates, monacolin J, which is the preferred precursor for Zocor synthesis [24]. Notably, a genetically engineered A. oryzae strain can reach monacolin J concentrations of about 5.5 g/L, which is considerably higher than has been achieved in heterologous hosts, such as S. cerevisiae (75 mg/L) or Pichia pastoris (renamed Komagataella phaffii; 600 mg/L) [25]. P. chrysogenum (P. rubens) is an antibiotics producer and the main cell factory used to produce penicillin and semisynthetic derivatives, with production levels up to 55 g/L [26].…”

Engineering cytochrome P450 enzyme systems for biomedical and biotechnological applications

“…3, compound 2) (34); and Aspergillus terreus LovA is responsible for biosynthesizing monacolin J acid (Fig. 3, compound 3), the precursor of a series of cholesterol-lowering statin drugs (35,36). The production of high value-added chemical intermediates from phenolic environmental pollutants has been achieved with P450 biodegradation systems (32,37,38), and soluble P450s have been used in bacterial cell libraries to mimic human P450 drug metabolic profiles (39,40).…”

Yeast transcriptional device libraries enable precise synthesis of value-added chemicals from methanol

“…Recently, we achieved an MJ production of 3.2 g/l by heterologous synthesis of this compound in P. pastoris using ethanol as a substrate ( 29 ). In addition, a high MJ production of 5.5 g/l from glucose achieved by rewiring metabolic pathways has been reported in the native fungus Aspergillus terreus ( 72 ). Therefore, there is still room for improvement in the synthesis of MJ using methanol as the sole carbon source.…”