Fig. 1.
Levodopa is deaminated by Clostridium sporogenes. a Full reductive deamination pathway of C. sporogenes is depicted resulting in the full deamination (R-propionic acid) of (non)-proteinogenic aromatic amino acids ((N)PAAAs). The red arrow indicates a disrupted deamination pathway of C. sporogenes, where the dehydratase subunit fldC is mutagenized, resulting in a pool of partially deaminated metabolites (R-lactic acid) by C. sporogenes. b HPLC-ED curves from supernatant of a C. sporogenes batch culture conversion of levodopa (3-(3,4-dihydroxyphenyl)alanine) over time. At the beginning of growth (timepoint 0 h), 100 μM of levodopa (blue) is added to the culture medium; the black line in the chromatogram depicts the control samples. In 24 h, levodopa is completely converted to DHPPA (3-(3,4-dihydroxyphenyl)propionic acid), the deaminated product of levodopa. Other aromatic amino acids from the medium, tryptophan and tyrosine (which are detectable with ED), are converted to the deaminated products IPA (indole-3-propionic acid) and 4-HPPA (3-(4-hydroxyphenyl)propionic acid). c Quantification (n = 3) of levodopa conversion to DHPPA by C. sporogenes wild type (also see Additional File 1: Table S1). d Analysis of the supernatant of CSΩfldC shows that levodopa is not deaminated to DHPPA but to its intermediate product DHPLA (3-(3,4-dihydroxyphenyl)lactic acid) within 24 h. Tryptophan and tyrosine are converted to their intermediates ILA (indole-3-lactic acid) and 4-HPLA (3-(4-hydroxyphenyl)lactic acid), respectively. e Quantification (n = 3) of levodopa conversion to DHPLA by C. sporogenes ΩfldC (also see Additional File 1: Table S1). All experiments were performed in 3 independent biological replicates, and means with error bars representing the SEM are depicted