| Affiliation: | 1. School of Chemical and Biological Engineering, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul, 08826 Republic of Korea;2. Institute of Molecular Biology and Genetics, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul, 08826 Republic of Korea
Institute of Engineering Research, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul, 08826 Republic of Korea;3. Institute of Molecular Biology and Genetics, Seoul National University, Gwanak-ro 1, Gwanak-gu, Seoul, 08826 Republic of Korea;4. Department of Biological and Chemical Engineering, Hongik University, Sejong-ro 2639, Jochiwon-eup, Sejong, 30016 Republic of Korea;5. Department of Environmental Engineering, Ajou University, World cup-ro 206, Yeongtong-gu, Suwon, 16499 Republic of Korea |
| Abstract: | Tryptophan halogenases are found in diverse organisms and catalyze regiospecific halogenation. They play an important role in the biosynthesis of halogenated indole alkaloids, which are biologically active and of therapeutic importance. Here, a tryptophan 6-halogenase (SatH) from Streptomyces albus was characterized by using a whole-cell reaction system in Escherichia coli. SatH showed substrate specificity for chloride and bromide ions, leading to regiospecific halogenation at the C6-position of l -tryptophan. In addition, SatH exhibited higher performance in bromination than that of previously reported tryptophan halogenases in the whole-cell reaction system. Through structure-based protein mutagenesis, it has been revealed that two consecutive residues, A78/V79 in SatH and G77/I78 in PyrH, are key determinants in the regioselectivity difference between tryptophan 6- and 5-halogenases. Substituting the AV with GI residues switched the regioselectivity of SatH by moving the orientation of tryptophan. These data contribute to an understanding of the key residues that determine the regioselectivity of tryptophan halogenases. |
