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Process development for efficient biosynthesis of l-DOPA with recombinant Escherichia coli harboring tyrosine phenol lyase from Fusobacterium nucleatum


The tyrosine phenol lyase (TPL) catalyzed synthesis of L-DOPA was regarded as one of the most economic route for L-DOPA synthesis. In our previous study, a novel TPL from Fusobacterium nucleatum (Fn-TPL) was exploited for efficient biosynthesis of L-DOPA. However, the catalytic efficiency decreased when the reaction system expanded from 100 mL to 1 L. As such, the bioprocess for scale-up production of L-DOPA was developed in this study. To increase the stability of substrate and product, as well as decrease the by-product formation, the optimum temperature and pH were determined to be 15 °C and pH 8.0, respectively. The initial concentration of pyrocatechol, pyruvate and ammonium acetate was fixed at 8, 5 and 77 g/L and a fed-batch approach was applied with sodium pyruvate, pyrocatechol and ammonium acetate fed in a concentration of 5, 5 and 3.5 g/L, respectively. In addition, L-DOPA crystals were exogenously added to inhibit cell encapsulation by the precipitated product. The final L-DOPA concentration reached higher than 120 g/L with pyrocatechol conversion more than 96% in a 15-L stirred tank, demonstrating the great potential of Fn-TPL for industrial production of L-DOPA.

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This study was financially supported by the Natural Science Foundation of Zhejiang Province (LQ17B060004).

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Correspondence to Ren-Chao Zheng.

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Tang, X., Liu, X., Suo, H. et al. Process development for efficient biosynthesis of l-DOPA with recombinant Escherichia coli harboring tyrosine phenol lyase from Fusobacterium nucleatum. Bioprocess Biosyst Eng 41, 1347–1354 (2018). https://doi.org/10.1007/s00449-018-1962-8

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  • L-DOPA
  • Tyrosine phenol lyase
  • Process development
  • Fed-batch reaction