As a kind of natural chemical, patchoulol is widely used in cosmetics industry. Compared with plant extraction, patchoulol production with microbial fermentation has great significance for saving cost and protecting environment. The solubility-enhancing tags were fused to patchoulol synthase PTS2 to improve soluble expression, and patchoulol production of strain Escherichia coli BL21/pMev-pET28a-PTS2T7A was increased by 39.74 %. Through the methods of homologous alignment, 3D structural modeling and analysis of surface amino acid, the amino-acid mutation sites of PTS2 were determined. Fermentation experiments using PTS2 mutants indicated that E457, K28, K136 and K343 were the key amino acid sites which can affect enzyme activity significantly. Furthermore, the eight genes of farnesyl pyrophosphate synthetic pathway were modularized, and strengthened by T7 promoter. Patchoulol production in the resulting strain BL21/pMev-T7M1M2+pET28a-PTS2 was increased by 55.95%. Finally, the optimal modification strategies were combined, and patchoulol production in the resulting strain BL21/pMev-T7M1M2+pET28a-PTS2T7A was cumulatively increased by 72.22%. The results provide reference for the production of patchoulol and other sesquiterpenes in E. coli.
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