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Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases

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Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases. / Srivastava, Prabhakar L; Johns, Sam T; Walters, Rebecca et al.
In: ACS Catalysis, Vol. 13, No. 21, 03.11.2023, p. 14199-14204.

Research output: Contribution to journal › Article › peer-review

HarvardHarvard

Srivastava, PL, Johns, ST, Walters, R, Miller, DJ, Van der Kamp, MW & Allemann, RK 2023, 'Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases', ACS Catalysis, vol. 13, no. 21, pp. 14199-14204. https://doi.org/10.1021/acscatal.3c03920

APA

Srivastava, P. L., Johns, S. T., Walters, R., Miller, D. J., Van der Kamp, M. W., & Allemann, R. K. (2023). Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases. ACS Catalysis, 13(21), 14199-14204. https://doi.org/10.1021/acscatal.3c03920

CBE

Srivastava PL, Johns ST, Walters R, Miller DJ, Van der Kamp MW, Allemann RK. 2023. Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases. ACS Catalysis. 13(21):14199-14204. https://doi.org/10.1021/acscatal.3c03920

MLA

Srivastava, Prabhakar L et al. "Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases". ACS Catalysis. 2023, 13(21). 14199-14204. https://doi.org/10.1021/acscatal.3c03920

VancouverVancouver

Srivastava PL, Johns ST, Walters R, Miller DJ, Van der Kamp MW, Allemann RK. Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases. ACS Catalysis. 2023 Nov 3;13(21):14199-14204. Epub 2023 Oct 20. doi: 10.1021/acscatal.3c03920

Author

Srivastava, Prabhakar L ; Johns, Sam T ; Walters, Rebecca et al. / Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases. In: ACS Catalysis. 2023 ; Vol. 13, No. 21. pp. 14199-14204.

RIS

TY - JOUR

T1 - Active Site Loop Engineering Abolishes Water Capture in Hydroxylating Sesquiterpene Synthases

AU - Srivastava, Prabhakar L

AU - Johns, Sam T

AU - Walters, Rebecca

AU - Miller, David J

AU - Van der Kamp, Marc W

AU - Allemann, Rudolf K

N1 - © 2023 The Authors. Published by American Chemical Society.

PY - 2023/11/3

Y1 - 2023/11/3

N2 - Terpene synthases (TS) catalyze complex reactions to produce a diverse array of terpene skeletons from linear isoprenyl diphosphates. Patchoulol synthase (PTS) from Pogostemon cablin converts farnesyl diphosphate into patchoulol. Using simulation-guided engineering, we obtained PTS variants that eliminate water capture. Further, we demonstrate that modifying the structurally conserved Hα-1 loop also reduces hydroxylation in PTS, as well as in germacradiene-11-ol synthase (Gd11olS), leading to cyclic neutral intermediates as products, including α-bulnesene (PTS) and isolepidozene (Gd11olS). Hα-1 loop modification could be a general strategy for engineering sesquiterpene synthases to produce complex cyclic hydrocarbons without the need for structure determination or modeling.

AB - Terpene synthases (TS) catalyze complex reactions to produce a diverse array of terpene skeletons from linear isoprenyl diphosphates. Patchoulol synthase (PTS) from Pogostemon cablin converts farnesyl diphosphate into patchoulol. Using simulation-guided engineering, we obtained PTS variants that eliminate water capture. Further, we demonstrate that modifying the structurally conserved Hα-1 loop also reduces hydroxylation in PTS, as well as in germacradiene-11-ol synthase (Gd11olS), leading to cyclic neutral intermediates as products, including α-bulnesene (PTS) and isolepidozene (Gd11olS). Hα-1 loop modification could be a general strategy for engineering sesquiterpene synthases to produce complex cyclic hydrocarbons without the need for structure determination or modeling.

U2 - 10.1021/acscatal.3c03920

DO - 10.1021/acscatal.3c03920

M3 - Article

C2 - 37942265

VL - 13

SP - 14199

EP - 14204

JO - ACS Catalysis

JF - ACS Catalysis

SN - 2155-5435

IS - 21

ER -