Engineering highly functional thermostable proteins using ancestral sequence reconstruction

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Engineering highly functional thermostable proteins using ancestral sequence reconstruction. / Gumulya, Yosephin; Baek, Jong-min; Wun, Shun-jie; Thomson, Raine E. S.; Harris, Kurt L.; Hunter, Dominic J. B.; Behrendorff, James B.Y.H.; Kulig, Justyna; Zheng, Shan; Wu, Xueming; Wu, Bin; Stok, Jeanette E.; De Voss, James J.; Schenk, Gerhard; Jurva, Ulrik; Andersson, Shalini; Isin, Emre M.; Bodén, Mikael; Guddat, Luke; Gillam, Elizabeth M. J.

In: Nature Catalysis, Vol. 1, No. 11, 2018, p. 878-888.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Gumulya, Y, Baek, J, Wun, S, Thomson, RES, Harris, KL, Hunter, DJB, Behrendorff, JBYH, Kulig, J, Zheng, S, Wu, X, Wu, B, Stok, JE, De Voss, JJ, Schenk, G, Jurva, U, Andersson, S, Isin, EM, Bodén, M, Guddat, L & Gillam, EMJ 2018, 'Engineering highly functional thermostable proteins using ancestral sequence reconstruction', Nature Catalysis, vol. 1, no. 11, pp. 878-888. https://doi.org/10.1038/s41929-018-0159-5

APA

Gumulya, Y., Baek, J., Wun, S., Thomson, R. E. S., Harris, K. L., Hunter, D. J. B., Behrendorff, J. B. Y. H., Kulig, J., Zheng, S., Wu, X., Wu, B., Stok, J. E., De Voss, J. J., Schenk, G., Jurva, U., Andersson, S., Isin, E. M., Bodén, M., Guddat, L., & Gillam, E. M. J. (2018). Engineering highly functional thermostable proteins using ancestral sequence reconstruction. Nature Catalysis, 1(11), 878-888. https://doi.org/10.1038/s41929-018-0159-5

Vancouver

Gumulya Y, Baek J, Wun S, Thomson RES, Harris KL, Hunter DJB et al. Engineering highly functional thermostable proteins using ancestral sequence reconstruction. Nature Catalysis. 2018;1(11):878-888. https://doi.org/10.1038/s41929-018-0159-5

Author

Gumulya, Yosephin ; Baek, Jong-min ; Wun, Shun-jie ; Thomson, Raine E. S. ; Harris, Kurt L. ; Hunter, Dominic J. B. ; Behrendorff, James B.Y.H. ; Kulig, Justyna ; Zheng, Shan ; Wu, Xueming ; Wu, Bin ; Stok, Jeanette E. ; De Voss, James J. ; Schenk, Gerhard ; Jurva, Ulrik ; Andersson, Shalini ; Isin, Emre M. ; Bodén, Mikael ; Guddat, Luke ; Gillam, Elizabeth M. J. / Engineering highly functional thermostable proteins using ancestral sequence reconstruction. In: Nature Catalysis. 2018 ; Vol. 1, No. 11. pp. 878-888.

Bibtex

@article{43c66336b762480a93bed53c4612114a,
title = "Engineering highly functional thermostable proteins using ancestral sequence reconstruction",
abstract = "Commercial biocatalysis requires robust enzymes that can withstand elevated temperatures and long incubations. Ancestral reconstruction has shown that pre-Cambrian enzymes were often much more thermostable than extant forms. Here, we resurrect ancestral enzymes that withstand ~30 °C higher temperatures and ≥100 times longer incubations than their extant forms. This is demonstrated on animal cytochromes P450 that stereo- and regioselectively functionalize unactivated C–H bonds for the synthesis of valuable chemicals, and bacterial ketol-acid reductoisomerases that are used to make butanol-based biofuels. The vertebrate CYP3 P450 ancestor showed a 60T50 of 66 °C and enhanced solvent tolerance compared with the human drug-metabolizing CYP3A4, yet comparable activity towards a similarly broad range of substrates. The ancestral ketol-acid reductoisomerase showed an eight-fold higher specific activity than the cognate Escherichia coli form at 25 °C, which increased 3.5-fold at 50 °C. Thus, thermostable proteins can be devised using sequence data alone from even recent ancestors.",
author = "Yosephin Gumulya and Jong-min Baek and Shun-jie Wun and Thomson, {Raine E. S.} and Harris, {Kurt L.} and Hunter, {Dominic J. B.} and Behrendorff, {James B.Y.H.} and Justyna Kulig and Shan Zheng and Xueming Wu and Bin Wu and Stok, {Jeanette E.} and {De Voss}, {James J.} and Gerhard Schenk and Ulrik Jurva and Shalini Andersson and Isin, {Emre M.} and Mikael Bod{\'e}n and Luke Guddat and Gillam, {Elizabeth M. J.}",
year = "2018",
doi = "10.1038/s41929-018-0159-5",
language = "English",
volume = "1",
pages = "878--888",
journal = "Nature Catalysis",
issn = "2520-1158",
publisher = "Macmillan Press",
number = "11",

}

RIS

TY - JOUR

T1 - Engineering highly functional thermostable proteins using ancestral sequence reconstruction

AU - Gumulya, Yosephin

AU - Baek, Jong-min

AU - Wun, Shun-jie

AU - Thomson, Raine E. S.

AU - Harris, Kurt L.

AU - Hunter, Dominic J. B.

AU - Behrendorff, James B.Y.H.

AU - Kulig, Justyna

AU - Zheng, Shan

AU - Wu, Xueming

AU - Wu, Bin

AU - Stok, Jeanette E.

AU - De Voss, James J.

AU - Schenk, Gerhard

AU - Jurva, Ulrik

AU - Andersson, Shalini

AU - Isin, Emre M.

AU - Bodén, Mikael

AU - Guddat, Luke

AU - Gillam, Elizabeth M. J.

PY - 2018

Y1 - 2018

N2 - Commercial biocatalysis requires robust enzymes that can withstand elevated temperatures and long incubations. Ancestral reconstruction has shown that pre-Cambrian enzymes were often much more thermostable than extant forms. Here, we resurrect ancestral enzymes that withstand ~30 °C higher temperatures and ≥100 times longer incubations than their extant forms. This is demonstrated on animal cytochromes P450 that stereo- and regioselectively functionalize unactivated C–H bonds for the synthesis of valuable chemicals, and bacterial ketol-acid reductoisomerases that are used to make butanol-based biofuels. The vertebrate CYP3 P450 ancestor showed a 60T50 of 66 °C and enhanced solvent tolerance compared with the human drug-metabolizing CYP3A4, yet comparable activity towards a similarly broad range of substrates. The ancestral ketol-acid reductoisomerase showed an eight-fold higher specific activity than the cognate Escherichia coli form at 25 °C, which increased 3.5-fold at 50 °C. Thus, thermostable proteins can be devised using sequence data alone from even recent ancestors.

AB - Commercial biocatalysis requires robust enzymes that can withstand elevated temperatures and long incubations. Ancestral reconstruction has shown that pre-Cambrian enzymes were often much more thermostable than extant forms. Here, we resurrect ancestral enzymes that withstand ~30 °C higher temperatures and ≥100 times longer incubations than their extant forms. This is demonstrated on animal cytochromes P450 that stereo- and regioselectively functionalize unactivated C–H bonds for the synthesis of valuable chemicals, and bacterial ketol-acid reductoisomerases that are used to make butanol-based biofuels. The vertebrate CYP3 P450 ancestor showed a 60T50 of 66 °C and enhanced solvent tolerance compared with the human drug-metabolizing CYP3A4, yet comparable activity towards a similarly broad range of substrates. The ancestral ketol-acid reductoisomerase showed an eight-fold higher specific activity than the cognate Escherichia coli form at 25 °C, which increased 3.5-fold at 50 °C. Thus, thermostable proteins can be devised using sequence data alone from even recent ancestors.

U2 - 10.1038/s41929-018-0159-5

DO - 10.1038/s41929-018-0159-5

M3 - Journal article

VL - 1

SP - 878

EP - 888

JO - Nature Catalysis

JF - Nature Catalysis

SN - 2520-1158

IS - 11

ER -

ID: 212167152