| PRAKINEE, Kridsadakorn, Aisaraphon PHINTHA, Surawit VISITSATTHAWONG, Narin LAWAN, Jeerus SUCHARITAKUL, Chadaporn KANTIWIRIYAWANITCH, Jiří DAMBORSKÝ, Penchit CHITNUMSUB, Van Pee KARL-HEINZ a Pimchai CHAIYEN. Mechanism-guided tunnel engineering to increase the efficiency of a flavin-dependent halogenase. NATURE CATALYSIS. ENGLAND: NATURE PORTFOLIO, 2022, roč. 5, č. 6, s. 534-544. ISSN 2520-1158. Dostupné z: https://dx.doi.org/10.1038/s41929-022-00800-8. |
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@article{1883608,
author = {Prakinee, Kridsadakorn and Phintha, Aisaraphon and Visitsatthawong, Surawit and Lawan, Narin and Sucharitakul, Jeerus and Kantiwiriyawanitch, Chadaporn and Damborský, Jiří and Chitnumsub, Penchit and KarlandHeinz, Van Pee and Chaiyen, Pimchai},
article_location = {ENGLAND},
article_number = {6},
doi = {http://dx.doi.org/10.1038/s41929-022-00800-8},
keywords = {TRYPTOPHAN 7-HALOGENASE; BIOCATALYTIC SCOPE; MOLECULAR-DYNAMICS; KINETIC MECHANISM; CHLORINATION; BIOSYNTHESIS; INTERMEDIATE; INSIGHTS; PHENOL; REBH},
language = {eng},
issn = {2520-1158},
journal = {NATURE CATALYSIS},
title = {Mechanism-guided tunnel engineering to increase the efficiency of a flavin-dependent halogenase},
url = {https://www.nature.com/articles/s41929-022-00800-8},
volume = {5},
year = {2022}
}
TY - JOUR
ID - 1883608
AU - Prakinee, Kridsadakorn - Phintha, Aisaraphon - Visitsatthawong, Surawit - Lawan, Narin - Sucharitakul, Jeerus - Kantiwiriyawanitch, Chadaporn - Damborský, Jiří - Chitnumsub, Penchit - Karl-Heinz, Van Pee - Chaiyen, Pimchai
PY - 2022
TI - Mechanism-guided tunnel engineering to increase the efficiency of a flavin-dependent halogenase
JF - NATURE CATALYSIS
VL - 5
IS - 6
SP - 534-544
EP - 534-544
PB - NATURE PORTFOLIO
SN - 25201158
KW - TRYPTOPHAN 7-HALOGENASE
KW - BIOCATALYTIC SCOPE
KW - MOLECULAR-DYNAMICS
KW - KINETIC MECHANISM
KW - CHLORINATION
KW - BIOSYNTHESIS
KW - INTERMEDIATE
KW - INSIGHTS
KW - PHENOL
KW - REBH
UR - https://www.nature.com/articles/s41929-022-00800-8
N2 - Although flavin-dependent halogenases (FDHs) are attractive for C-H bond activation, their applications are limited due to low turnover and stability. We have previously shown that leakage of a halogenating intermediate, hypohalous acid (HOX), causes FDHs to be inefficient by lessening halogenation yield. Here we employed a mechanism-guided semi-rational approach to engineer the intermediate transfer tunnel connecting two active sites of tryptophan 6-halogenase (Thal). This Thal-V82I variant generates less HOX leakage and possesses multiple catalytic improvements such as faster halogenation, broader substrate utilization, and greater thermostability and pH tolerance compared with the wildtype Thal. Stopped-flow and rapid quench kinetics analyses indicated that rate constants of halogenation and flavin oxidation are faster for Thal-V82I. Molecular dynamics simulations revealed that the V82I substitution introduces hydrophobic interactions which regulate tunnel dynamics to accommodate HOX and cause rearrangement of water networks, allowing better use of various substrates than the wildtype. Our approach demonstrates that an in-depth understanding of reaction mechanisms is valuable for improving efficiency of FDHs.
ER -
PRAKINEE, Kridsadakorn, Aisaraphon PHINTHA, Surawit VISITSATTHAWONG, Narin LAWAN, Jeerus SUCHARITAKUL, Chadaporn KANTIWIRIYAWANITCH, Jiří DAMBORSKÝ, Penchit CHITNUMSUB, Van Pee KARL-HEINZ a Pimchai CHAIYEN. Mechanism-guided tunnel engineering to increase the efficiency of a flavin-dependent halogenase. \textit{NATURE CATALYSIS}. ENGLAND: NATURE PORTFOLIO, 2022, roč.~5, č.~6, s.~534-544. ISSN~2520-1158. Dostupné z: https://dx.doi.org/10.1038/s41929-022-00800-8.
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