Production of L-DOPA and dopamine in recombinant bacteria bearing the Vitreoscilla hemoglobin gene.
Given the well-established beneficial effects of Vitreoscilla hemoglobin (VHb) on heterologous organisms, the potential of this protein for the production of L-DOPA and dopamine in two bacteria, Citrobacter freundii and Erwinia herbicola, was investigated. The constructed recombinants bearing the VHb gene (vgb(+)) had substantially higher levels of cytoplasmic L-DOPA (112 mg/L for C. freundii and 97 mg/L for E. herbicola) than their respective hosts (30.4 and 33.8 mg/L) and the vgb(-) control strains (35.6 and 35.8 mg/L). Further, the vgb(+) recombinants of C. freundii and E. herbicola had 20-fold and about two orders of magnitude higher dopamine levels than their hosts, repectively. The activity of tyrosine phenol-lyase, the enzyme converting L-tyrosine to L-DOPA, was well-correlated to cytoplasmic L-DOPA levels. As cultures aged, higher tyrosine phenol-lyase activity of the vgb(+) strains was more apparent.
Kurt
Asli Giray
AG
Department of Biology, Inonu University, Malatya, Turkey.
Aytan
Emel
E
Ozer
Ufuk
U
Ates
Burhan
B
Geckil
Hikmet
H
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016971:Citrobacter freundii / Q000235:genetics / Q000378:metabolism; D004298:Dopamine / Q000096:biosynthesis* / Q000235:genetics; D004885:Erwinia / Q000235:genetics / Q000378:metabolism; D006420:Hemeproteins / Q000235:genetics / Q000378:metabolism*; D007980:Levodopa / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D054793:Truncated Hemoglobins / Q000235:genetics / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism; D014447:Tyrosine Phenol-Lyase / Q000378:metabolism
D001426:Bacterial Proteins; D006420:Hemeproteins; D011994:Recombinant Proteins; D054793:Truncated Hemoglobins; C100771:Vgb protein, Bacteria; C082613:hemoglobin protein, Vitreoscilla; D014443:Tyrosine; D007980:Levodopa; D014447:Tyrosine Phenol-Lyase; D004298:Dopamine