Structural Dynamics and Catalytic Properties of a Multimodular Xanthanase

Olga V. Moroz; Pernille F. Jensen; Sean P. Mcdonald; Nicholas Mcgregor; Elena Blagova; Gerard Comamala; Dorotea R. Segura; Lars Anderson; Santhosh M. Vasu; Vasudeva P. Rao; Lars Giger; Trine Holst Sørensen; Rune Nygaard Monrad; Allan Svendsen; Jens E. Nielsen; Bernard Henrissat; Gideon J. Davies; Harry Brumer; Kasper D. Rand; Keith S. Wilson

doi:10.1021/acscatal.8b00666

Structural Dynamics and Catalytic Properties of a Multimodular Xanthanase

Olga V. Moroz, Pernille F. Jensen, Sean P. Mcdonald, Nicholas Mcgregor, Elena Blagova, Gerard Comamala, Dorotea R. Segura, Lars Anderson, Santhosh M. Vasu, Vasudeva P. Rao, Lars Giger, Trine Holst Sørensen, Rune Nygaard Monrad, Allan Svendsen, Jens E. Nielsen, Bernard Henrissat, Gideon J. Davies, Harry Brumer, Kasper D. Rand, Keith S. Wilson

5 Citationer (Scopus)

Abstract

The precise catalytic strategies used for the breakdown of the complex bacterial polysaccharide xanthan, an increasingly frequent component of processed human foodstuffs, have remained a mystery. Here, we present characterization of an endo-xanthanase from Paenibacillus nanensis. We show that it is a CAZy family 9 glycoside hydrolase (GH9) responsible for the hydrolysis of the xanthan backbone capable of generating tetrameric xanthan oligosaccharides from polysaccharide lyase family 8 (PL8) xanthan lyase-treated xanthan. Three-dimensional structure determination reveals a complex multimodular enzyme in which a catalytic (α/α)₆ barrel is flanked by an N-terminal "immunoglobulin-like" (Ig-like) domain (frequently found in GH9 enzymes) and by four additional C-terminal all β-sheet domains that have very few homologues in sequence databases and at least one of which functions as a new xanthan-binding domain, now termed CBM84. The solution-phase conformation and dynamics of the enzyme in the native calcium-bound state and in the absence of calcium were probed experimentally by hydrogen/deuterium exchange mass spectrometry. Measured conformational dynamics were used to guide the protein engineering of enzyme variants with increased stability in the absence of calcium; a property of interest for the potential use of the enzyme in cleaning detergents. The ability of hydrogen/deuterium exchange mass spectrometry to pinpoint dynamic regions of a protein under stress (e.g., removal of calcium ions) makes this technology a strong tool for improving protein catalyst properties by informed engineering.

Originalsprog	Engelsk
Tidsskrift	ACS Catalysis
Vol/bind	8
Udgave nummer	7
Sider (fra-til)	6021-6034
ISSN	2155-5435
DOI	https://doi.org/10.1021/acscatal.8b00666
Status	Udgivet - 6 jul. 2018

Adgang til dokumentet

10.1021/acscatal.8b00666

Citationsformater

Moroz, O. V., Jensen, P. F., Mcdonald, S. P., Mcgregor, N., Blagova, E., Comamala, G., Segura, D. R., Anderson, L., Vasu, S. M., Rao, V. P., Giger, L., Sørensen, T. H., Monrad, R. N., Svendsen, A., Nielsen, J. E., Henrissat, B., Davies, G. J., Brumer, H., Rand, K. D., & Wilson, K. S. (2018). Structural Dynamics and Catalytic Properties of a Multimodular Xanthanase. ACS Catalysis, 8(7), 6021-6034. https://doi.org/10.1021/acscatal.8b00666

Moroz, OV, Jensen, PF, Mcdonald, SP, Mcgregor, N, Blagova, E, Comamala, G, Segura, DR, Anderson, L, Vasu, SM, Rao, VP, Giger, L, Sørensen, TH, Monrad, RN, Svendsen, A, Nielsen, JE, Henrissat, B, Davies, GJ, Brumer, H, Rand, KD & Wilson, KS 2018, 'Structural Dynamics and Catalytic Properties of a Multimodular Xanthanase', ACS Catalysis, bind 8, nr. 7, s. 6021-6034. https://doi.org/10.1021/acscatal.8b00666

@article{c856a7a0b827413f9a01a3aa6ef5c052,

title = "Structural Dynamics and Catalytic Properties of a Multimodular Xanthanase",

abstract = "The precise catalytic strategies used for the breakdown of the complex bacterial polysaccharide xanthan, an increasingly frequent component of processed human foodstuffs, have remained a mystery. Here, we present characterization of an endo-xanthanase from Paenibacillus nanensis. We show that it is a CAZy family 9 glycoside hydrolase (GH9) responsible for the hydrolysis of the xanthan backbone capable of generating tetrameric xanthan oligosaccharides from polysaccharide lyase family 8 (PL8) xanthan lyase-treated xanthan. Three-dimensional structure determination reveals a complex multimodular enzyme in which a catalytic (α/α)6 barrel is flanked by an N-terminal {"}immunoglobulin-like{"} (Ig-like) domain (frequently found in GH9 enzymes) and by four additional C-terminal all β-sheet domains that have very few homologues in sequence databases and at least one of which functions as a new xanthan-binding domain, now termed CBM84. The solution-phase conformation and dynamics of the enzyme in the native calcium-bound state and in the absence of calcium were probed experimentally by hydrogen/deuterium exchange mass spectrometry. Measured conformational dynamics were used to guide the protein engineering of enzyme variants with increased stability in the absence of calcium; a property of interest for the potential use of the enzyme in cleaning detergents. The ability of hydrogen/deuterium exchange mass spectrometry to pinpoint dynamic regions of a protein under stress (e.g., removal of calcium ions) makes this technology a strong tool for improving protein catalyst properties by informed engineering.",

author = "Moroz, {Olga V.} and Jensen, {Pernille F.} and Mcdonald, {Sean P.} and Nicholas Mcgregor and Elena Blagova and Gerard Comamala and Segura, {Dorotea R.} and Lars Anderson and Vasu, {Santhosh M.} and Rao, {Vasudeva P.} and Lars Giger and S{\o}rensen, {Trine Holst} and Monrad, {Rune Nygaard} and Allan Svendsen and Nielsen, {Jens E.} and Bernard Henrissat and Davies, {Gideon J.} and Harry Brumer and Rand, {Kasper D.} and Wilson, {Keith S.}",

year = "2018",

month = jul,

day = "6",

doi = "10.1021/acscatal.8b00666",

language = "English",

volume = "8",

pages = "6021--6034",

journal = "ACS Catalysis",

issn = "2155-5435",

publisher = "American Chemical Society",

number = "7",

}

TY - JOUR

T1 - Structural Dynamics and Catalytic Properties of a Multimodular Xanthanase

AU - Moroz, Olga V.

AU - Jensen, Pernille F.

AU - Mcdonald, Sean P.

AU - Mcgregor, Nicholas

AU - Blagova, Elena

AU - Comamala, Gerard

AU - Segura, Dorotea R.

AU - Anderson, Lars

AU - Vasu, Santhosh M.

AU - Rao, Vasudeva P.

AU - Giger, Lars

AU - Sørensen, Trine Holst

AU - Monrad, Rune Nygaard

AU - Svendsen, Allan

AU - Nielsen, Jens E.

AU - Henrissat, Bernard

AU - Davies, Gideon J.

AU - Brumer, Harry

AU - Rand, Kasper D.

AU - Wilson, Keith S.

PY - 2018/7/6

Y1 - 2018/7/6

N2 - The precise catalytic strategies used for the breakdown of the complex bacterial polysaccharide xanthan, an increasingly frequent component of processed human foodstuffs, have remained a mystery. Here, we present characterization of an endo-xanthanase from Paenibacillus nanensis. We show that it is a CAZy family 9 glycoside hydrolase (GH9) responsible for the hydrolysis of the xanthan backbone capable of generating tetrameric xanthan oligosaccharides from polysaccharide lyase family 8 (PL8) xanthan lyase-treated xanthan. Three-dimensional structure determination reveals a complex multimodular enzyme in which a catalytic (α/α)6 barrel is flanked by an N-terminal "immunoglobulin-like" (Ig-like) domain (frequently found in GH9 enzymes) and by four additional C-terminal all β-sheet domains that have very few homologues in sequence databases and at least one of which functions as a new xanthan-binding domain, now termed CBM84. The solution-phase conformation and dynamics of the enzyme in the native calcium-bound state and in the absence of calcium were probed experimentally by hydrogen/deuterium exchange mass spectrometry. Measured conformational dynamics were used to guide the protein engineering of enzyme variants with increased stability in the absence of calcium; a property of interest for the potential use of the enzyme in cleaning detergents. The ability of hydrogen/deuterium exchange mass spectrometry to pinpoint dynamic regions of a protein under stress (e.g., removal of calcium ions) makes this technology a strong tool for improving protein catalyst properties by informed engineering.

AB - The precise catalytic strategies used for the breakdown of the complex bacterial polysaccharide xanthan, an increasingly frequent component of processed human foodstuffs, have remained a mystery. Here, we present characterization of an endo-xanthanase from Paenibacillus nanensis. We show that it is a CAZy family 9 glycoside hydrolase (GH9) responsible for the hydrolysis of the xanthan backbone capable of generating tetrameric xanthan oligosaccharides from polysaccharide lyase family 8 (PL8) xanthan lyase-treated xanthan. Three-dimensional structure determination reveals a complex multimodular enzyme in which a catalytic (α/α)6 barrel is flanked by an N-terminal "immunoglobulin-like" (Ig-like) domain (frequently found in GH9 enzymes) and by four additional C-terminal all β-sheet domains that have very few homologues in sequence databases and at least one of which functions as a new xanthan-binding domain, now termed CBM84. The solution-phase conformation and dynamics of the enzyme in the native calcium-bound state and in the absence of calcium were probed experimentally by hydrogen/deuterium exchange mass spectrometry. Measured conformational dynamics were used to guide the protein engineering of enzyme variants with increased stability in the absence of calcium; a property of interest for the potential use of the enzyme in cleaning detergents. The ability of hydrogen/deuterium exchange mass spectrometry to pinpoint dynamic regions of a protein under stress (e.g., removal of calcium ions) makes this technology a strong tool for improving protein catalyst properties by informed engineering.

U2 - 10.1021/acscatal.8b00666

DO - 10.1021/acscatal.8b00666

M3 - Journal article

SN - 2155-5435

VL - 8

SP - 6021

EP - 6034

JO - ACS Catalysis

JF - ACS Catalysis

IS - 7

ER -

Structural Dynamics and Catalytic Properties of a Multimodular Xanthanase

Abstract

Adgang til dokumentet

Fingeraftryk

Citationsformater