Improved taxadiene production by optimizing DXS expression and fusing short-chain prenyltransferases

• Published in: New biotechnology 2024-11, Vol.83, p.66-73
• Main Authors: He, Siqi, Bekhof, Anne-Sophie M.W., Popova, Eli Z., van Merkerk, Ronald, Quax, Wim J.
• Format: Article
• Language: English
• Subjects: Alkenes - metabolism; antineoplastic agents; Bacillus subtilis; biosynthesis; biotechnology; dimethylallyltranstransferase; Dimethylallyltranstransferase - genetics; Dimethylallyltranstransferase - metabolism; Diterpenes - metabolism; Escherichia coli - genetics; Escherichia coli - metabolism; genes; paclitaxel; Pentosyltransferases - biosynthesis; Pentosyltransferases - genetics; Pentosyltransferases - metabolism; Prenyltransferase; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Taxadiene; Transferases
• ISSN: 1871-6784; 1876-4347; 1876-4347
• DOI: 10.1016/j.nbt.2024.06.007

This study highlights the significance of overexpressing 1-deoxy-d-xylulose-5-phosphate synthase (DXS) from the MEP (methylerythritol 4-phosphate) pathway, in addition to short-chain prenyltransferase fusions for the improved production of the diterpene, taxa-4,11-diene, the first committed intermediate in the production of anti-cancer drug paclitaxel. The results showed that the strain which has (i) the taxadiene synthase (txs) gene integrated into the genome, (ii) the MEP pathway genes overexpressed, (iii) the fpps-crtE prenyltransferases fusion protein and (iv) additional expression of 1-deoxy-d-xylulose-5-phosphate synthase (DXS), yielded the highest production of taxa-4,11-diene at 390 mg/L (26 mg/L/OD600). This represents a thirteen-fold increase compared to the highest reported concentration in B. subtilis. The focus on additional overexpression of DXS and utilizing short-chain prenyltransferase fusions underscores their pivotal role in achieving significant titer improvements in terpene biosynthesis. [Display omitted] •Utilizing Bacillus subtilis as a cell factory for the production of taxadiene.•Additional expression of DXS from MEP pathway can boost taxadiene production.•Application of fused prenyltransferases can enhance the yield of taxadiene.•The highest taxadiene production so far in B. subtilis has been achieved.