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

Pernille Foged Jensen; Anders Kadziola; Gerard Comamala; Dorotea R Segura; Lars Anderson; Jens-Christian N Poulsen; Kim Krighaar Rasmussen; Shilpi Agarwal; Rajendra K Sainathan; Rune Nygaard Monrad; Allan Svendsen; Jens Erik Nielsen; Leila Lo Leggio; Kasper D Rand

doi:10.1016/j.chembiol.2018.10.016

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

Pernille Foged Jensen, Anders Kadziola, Gerard Comamala, Dorotea R Segura, Lars Anderson, Jens-Christian N Poulsen, Kim Krighaar Rasmussen, Shilpi Agarwal, Rajendra K Sainathan, Rune Nygaard Monrad, Allan Svendsen, Jens Erik Nielsen, Leila Lo Leggio, Kasper D Rand

3 Citations (Scopus)

Abstract

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.

Original language	English
Journal	Cell Chemical Biology
Volume	26
Issue number	2
Pages (from-to)	191-202.e1-e6
Number of pages	19
ISSN	2451-9448
DOIs	https://doi.org/10.1016/j.chembiol.2018.10.016
Publication status	Published - 21 Feb 2019

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Jensen, P. F., Kadziola, A., Comamala, G., Segura, D. R., Anderson, L., Poulsen, J-C. N., Rasmussen, K. K., Agarwal, S., Sainathan, R. K., Monrad, R. N., Svendsen, A., Nielsen, J. E., Lo Leggio, L., & Rand, K. D. (2019). Structure and Dynamics of a Promiscuous Xanthan Lyase from Paenibacillus nanensis and the Design of Variants with Increased Stability and Activity. Cell Chemical Biology, 26(2), 191-202.e1-e6. https://doi.org/10.1016/j.chembiol.2018.10.016

Structure and Dynamics of a Promiscuous Xanthan Lyase from Paenibacillus nanensis and the Design of Variants with Increased Stability and Activity. / Jensen, Pernille Foged; Kadziola, Anders; Comamala, Gerard et al.

In: Cell Chemical Biology, Vol. 26, No. 2, 21.02.2019, p. 191-202.e1-e6.

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

Jensen, PF, Kadziola, A, Comamala, G, Segura, DR, Anderson, L, Poulsen, J-CN, Rasmussen, KK, Agarwal, S, Sainathan, RK, Monrad, RN, Svendsen, A, Nielsen, JE, Lo Leggio, L & Rand, KD 2019, 'Structure and Dynamics of a Promiscuous Xanthan Lyase from Paenibacillus nanensis and the Design of Variants with Increased Stability and Activity', Cell Chemical Biology, vol. 26, no. 2, pp. 191-202.e1-e6. https://doi.org/10.1016/j.chembiol.2018.10.016

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abstract = " 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. ",

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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

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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 - Chemistry and Biology

JF - Chemistry and Biology

IS - 2

ER -

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

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