Redox self-sufficient biocatalyst system for conversion of 3,4-Dihydroxyphenyl-L-alanine into (R)- or (S)-3,4-Dihydroxyphenyllactic acid.
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.
['Xiong|Tianzhen|T|', 'Jiang|Jing|J|', 'Bai|Yajun|Y|', 'Fan|Tai-Ping|TP|', 'Zhao|Ye|Y|', 'Zheng|Xiaohui|X|', 'Cai|Yujie|Y|']
D000409:Alanine D055162:Biocatalysis D004926:Escherichia coli D007773:Lactates D010084:Oxidation-Reduction
Self-sufficient 3,4-Dihydroxyphenyl-L-alanine (S)-3,4-Dihydroxyphenyllactic acid (R)-3,4-Dihydroxyphenyllactic acid Whole-cell biotransformation