Redox self-sufficient biocatalyst system for conversion of 3,4-Dihydroxyphenyl-l-alanine into (R)- or (S)-3,4-Dihydroxyphenyllactic acid
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We developed an efficient multi-enzyme cascade reaction to produce (R)- or (S)-3,4-Dihydroxyphenyllactic acid [(R)- or (S)-Danshensu, (R)- or (S)-DSS] from 3,4-Dihydroxyphenyl-l-alanine (l-DOPA) in Escherichia coli by introducing tyrosine aminotransferase (tyrB), glutamate dehydrogenase (cdgdh) and d-aromatic lactate dehydrogenase (csldhD) or l-aromatic lactate dehydrogenase (tcldhL). First, the genes in the pathway were overexpressed and fine-tuned for (R)- or (S)-DSS production. The resulting strain, E. coli TGL 2.1 and E. coli TGL 2.2, which overexpressed tyrB with the stronger T7 promoter and cdgdh, csldhD or tcldhL with the weaker Trc promoter, E. coli TGL 2.1 yielded 57% increase in (R)-DSS production: 59.8 ± 2.9 mM. Meanwhile, E. coli TGL 2.2 yielded 54% increase in (S)-DSS production: 52.2 ± 2.4 mM. The optimal concentration of L-glutamate was found to be 20 mM for production of (R)- or (S)-DSS. Finally, l-DOPA were transformed into (R)- or (S)-DSS with an excellent enantiopure form (enantiomeric excess > 99.99%) and productivity of 6.61 mM/h and 4.48 mM/h, respectively.
Keywords(R)-3,4-Dihydroxyphenyllactic acid (S)-3,4-Dihydroxyphenyllactic acid 3,4-Dihydroxyphenyl-l-alanine Self-sufficient Whole-cell biotransformation
We thank the National Key Scientific Instrument and Equipment Development Project of China (2013YQ17052504), national first-class discipline program of Light Industry Technology and Engineering (LITE2108-04), Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX19_1841), the Program for Changjiang Scholars and Innovative Research Team in the University of Ministry of Education of China (IRT_15R55), the seventh group of Hundred-Talent Program of Shanxi Province (2015), and The Key Project of Research and Development Plan of Shaanxi (2017ZDCXL-SF-01-02-01) for financial support.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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