Publication: Creating Flavin Reductase Variants with Thermostable and Solvent-Tolerant Properties by Rational-Design Engineering
Issued Date
2020-05-15
Resource Type
ISSN
14397633
14394227
14394227
Other identifier(s)
2-s2.0-85084935248
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Mahidol University
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SCOPUS
Bibliographic Citation
ChemBioChem. Vol.21, No.10 (2020), 1481-1491
Suggested Citation
Somchart Maenpuen, Vinutsada Pongsupasa, Wiranee Pensook, Piyanuch Anuwan, Napatsorn Kraivisitkul, Chatchadaporn Pinthong, Jittima Phonbuppha, Thikumporn Luanloet, Hein J. Wijma, Marco W. Fraaije, Narin Lawan, Pimchai Chaiyen, Thanyaporn Wongnate Creating Flavin Reductase Variants with Thermostable and Solvent-Tolerant Properties by Rational-Design Engineering. ChemBioChem. Vol.21, No.10 (2020), 1481-1491. doi:10.1002/cbic.201900737 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/56111
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Title
Creating Flavin Reductase Variants with Thermostable and Solvent-Tolerant Properties by Rational-Design Engineering
Abstract
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim We have employed computational approaches—FireProt and FRESCO—to predict thermostable variants of the reductase component (C1) of (4-hydroxyphenyl)acetate 3-hydroxylase. With the additional aid of experimental results, two C1 variants, A166L and A58P, were identified as thermotolerant enzymes, with thermostability improvements of 2.6–5.6 °C and increased catalytic efficiency of 2- to 3.5-fold. After heat treatment at 45 °C, both of the thermostable C1 variants remain active and generate reduced flavin mononucleotide (FMNH−) for reactions catalyzed by bacterial luciferase and by the monooxygenase C2 more efficiently than the wild type (WT). In addition to thermotolerance, the A166L and A58P variants also exhibited solvent tolerance. Molecular dynamics (MD) simulations (6 ns) at 300–500 K indicated that mutation of A166 to L and of A58 to P resulted in structural changes with increased stabilization of hydrophobic interactions, and thus in improved thermostability. Our findings demonstrated that improvements in the thermostability of C1 enzyme can lead to broad-spectrum uses of C1 as a redox biocatalyst for future industrial applications.
