Structure and Dynamics of a Promiscuous Xanthan Lyase from Paenibacillus nanensis and the Design of Variants with Increased Stability and Activity

TY - JOUR

T1 - Structure and Dynamics of a Promiscuous Xanthan Lyase from Paenibacillus nanensis and the Design of Variants with Increased Stability and Activity

AU - Jensen, Pernille Foged

AU - Kadziola, Anders

AU - Comamala, Gerard

AU - Segura, Dorotea R

AU - Anderson, Lars

AU - Poulsen, Jens-Christian N

AU - Rasmussen, Kim Krighaar

AU - Agarwal, Shilpi

AU - Sainathan, Rajendra K

AU - Monrad, Rune Nygaard

AU - Svendsen, Allan

AU - Nielsen, Jens Erik

AU - Lo Leggio, Leila

AU - Rand, Kasper D

N1 - Copyright © 2018 Elsevier Ltd. All rights reserved.

PY - 2019/2/21

Y1 - 2019/2/21

N2 - We have characterized the structure and dynamics of the carbohydrate-modifying enzyme Paenibacillus nanensis xanthan lyase (PXL) involved in the degradation of xanthan by X-ray crystallography, small-angle X-ray scattering, and hydrogen/deuterium exchange mass spectrometry. Unlike other xanthan lyases, PXL is specific for both unmodified mannose and pyruvylated mannose, which we find is correlated with structural differences in the substrate binding groove. The structure of the full-length enzyme reveals two additional C-terminal modules, one of which belongs to a new non-catalytic carbohydrate binding module family. Ca 2+ are critical for the activity and conformation of PXL, and we show that their removal by chelating agents results in localized destabilization/unfolding of particularly the C-terminal modules. We use the structure and the revealed impact of Ca 2+ coordination on conformational dynamics to guide the engineering of PXL variants with increased activity and stability in a chelating environment, thus expanding the possibilities for industrial applications of PXL. Jensen et al. describe the full-length structure and solution-phase dynamics of a xanthan lyase (PXL), an enzyme participating in degradation of the industrial heteropolysaccharide, xanthan. Insights into the structure and the impact of chelating agents on PXL dynamics are used to produce PXL variants with improved activity and stability.

AB - We have characterized the structure and dynamics of the carbohydrate-modifying enzyme Paenibacillus nanensis xanthan lyase (PXL) involved in the degradation of xanthan by X-ray crystallography, small-angle X-ray scattering, and hydrogen/deuterium exchange mass spectrometry. Unlike other xanthan lyases, PXL is specific for both unmodified mannose and pyruvylated mannose, which we find is correlated with structural differences in the substrate binding groove. The structure of the full-length enzyme reveals two additional C-terminal modules, one of which belongs to a new non-catalytic carbohydrate binding module family. Ca 2+ are critical for the activity and conformation of PXL, and we show that their removal by chelating agents results in localized destabilization/unfolding of particularly the C-terminal modules. We use the structure and the revealed impact of Ca 2+ coordination on conformational dynamics to guide the engineering of PXL variants with increased activity and stability in a chelating environment, thus expanding the possibilities for industrial applications of PXL. Jensen et al. describe the full-length structure and solution-phase dynamics of a xanthan lyase (PXL), an enzyme participating in degradation of the industrial heteropolysaccharide, xanthan. Insights into the structure and the impact of chelating agents on PXL dynamics are used to produce PXL variants with improved activity and stability.

U2 - 10.1016/j.chembiol.2018.10.016

DO - 10.1016/j.chembiol.2018.10.016

M3 - Journal article

C2 - 30503284

SN - 2451-9448

VL - 26

SP - 191-202.e1-e6

JO - Cell Chemical Biology

JF - Cell Chemical Biology

IS - 2

ER -