Biosynthesis of D-danshensu from L-DOPA using engineered Escherichia coli whole cells
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D-Danshensu (D-DSS), a traditional Chinese medicine, is used to treat cardiovascular and cerebrovascular diseases. However, current isolation protocols for D-DSS both natural and synthetic are not ideal; therefore, in this study, we have developed a whole-cell biotransformation method to produce D-DSS from L-DOPA. This was done by co-expressing L-amino acid deaminase (aadL), D-lactate dehydrogenase (ldhD), and glucose dehydrogenase (gdh). To begin to optimize the production of D-DSS, varying copy number plasmids were used to express each of the required genes. The resulting strain, Escherichia coli ALG7, which strongly overexpressed aadL, ldhD, and weakly overexpressed gdh, yielded a 378% increase in D-DSS production compared to E. coli ALG1. Furthermore, the optimal reaction conditions for the production of D-DSS were found to be a pH of 7.5, temperature at 35 °C, and 50 g/L wet cells for 12 h. Under these optimized conditions, the D-DSS amount achieved 119.1 mM with an excellent ee (> 99.9%) and a productivity of 9.9 mM/h.
KeywordsD-Danshensu Escherichia coli L-DOPA One-pot synthesis Whole-cell biotransformation
This study received financial support from the National Key Scientific Instrument and Equipment Development Project of China (2013YQ17052504), the national first-class discipline program of Light Industry Technology and Engineering (LITE2108-04), 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 Shaanxi Province (2015), and The Key Project of Research and Development Plan of Shaanxi (2017ZDCXL-SF-01-02-01).
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflict of interest.
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