Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases. / Tandrup, Tobias; Tryfona, Theodora; Frandsen, Kristian Erik Høpfner; Johansen, Katja Salomon; Dupree, Paul; Lo Leggio, Leila.

In: Biochemistry, Vol. 59, No. 36, 2020, p. 3347-3358.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Tandrup, T, Tryfona, T, Frandsen, KEH, Johansen, KS, Dupree, P & Lo Leggio, L 2020, 'Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases', Biochemistry, vol. 59, no. 36, pp. 3347-3358. https://doi.org/10.1021/acs.biochem.0c00312

APA

Tandrup, T., Tryfona, T., Frandsen, K. E. H., Johansen, K. S., Dupree, P., & Lo Leggio, L. (2020). Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases. Biochemistry, 59(36), 3347-3358. https://doi.org/10.1021/acs.biochem.0c00312

Vancouver

Tandrup T, Tryfona T, Frandsen KEH, Johansen KS, Dupree P, Lo Leggio L. Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases. Biochemistry. 2020;59(36):3347-3358. https://doi.org/10.1021/acs.biochem.0c00312

Author

Tandrup, Tobias ; Tryfona, Theodora ; Frandsen, Kristian Erik Høpfner ; Johansen, Katja Salomon ; Dupree, Paul ; Lo Leggio, Leila. / Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases. In: Biochemistry. 2020 ; Vol. 59, No. 36. pp. 3347-3358.

Bibtex

@article{50281ab32b3b4114894360576af05ade,
title = "Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases",
abstract = "Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that cleave polysaccharide substrates oxidatively. First discovered because of their action on recalcitrant crystalline substrates (chitin and cellulose), a number of LPMOs are now reported to act on soluble substrates, including oligosaccharides. However, crystallographic complexes with oligosaccharides have been reported for only a single LPMO so far, an enzyme from the basidiomycete fungus Lentinus similis (LsAA9_A). Here we present a more detailed comparative study of LsAA9_A and an LPMO from the ascomycete fungus Collariella virescens (CvAA9_A) with which it shares 41.5{\%} sequence identity. LsAA9_A is considerably more thermostable than CvAA9_A, and the structural basis for the difference has been investigated. We have compared the patterns of oligosaccharide cleavage and the patterns of binding in several new crystal structures explaining the basis for the product preferences of the two enzymes. Obtaining structural information about complexes of LPMOs with carbohydrates has proven to be very difficult in general judging from the structures reported in the literature thus far, and this can be attributed only partly to the low affinity for small substrates. We have thus evaluated the use of differential scanning fluorimetry as a guide to obtaining complex structures. Furthermore, an analysis of crystal packing of LPMOs and glycoside hydrolases corroborates the hypothesis that active site occlusion is a very significant problem for LPMO-substrate interaction analysis by crystallography, due to their relatively flat and extended substrate binding sites.",
author = "Tobias Tandrup and Theodora Tryfona and Frandsen, {Kristian Erik H{\o}pfner} and Johansen, {Katja Salomon} and Paul Dupree and {Lo Leggio}, Leila",
year = "2020",
doi = "10.1021/acs.biochem.0c00312",
language = "English",
volume = "59",
pages = "3347--3358",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "36",

}

RIS

TY - JOUR

T1 - Oligosaccharide Binding and Thermostability of Two Related AA9 Lytic Polysaccharide Monooxygenases

AU - Tandrup, Tobias

AU - Tryfona, Theodora

AU - Frandsen, Kristian Erik Høpfner

AU - Johansen, Katja Salomon

AU - Dupree, Paul

AU - Lo Leggio, Leila

PY - 2020

Y1 - 2020

N2 - Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that cleave polysaccharide substrates oxidatively. First discovered because of their action on recalcitrant crystalline substrates (chitin and cellulose), a number of LPMOs are now reported to act on soluble substrates, including oligosaccharides. However, crystallographic complexes with oligosaccharides have been reported for only a single LPMO so far, an enzyme from the basidiomycete fungus Lentinus similis (LsAA9_A). Here we present a more detailed comparative study of LsAA9_A and an LPMO from the ascomycete fungus Collariella virescens (CvAA9_A) with which it shares 41.5% sequence identity. LsAA9_A is considerably more thermostable than CvAA9_A, and the structural basis for the difference has been investigated. We have compared the patterns of oligosaccharide cleavage and the patterns of binding in several new crystal structures explaining the basis for the product preferences of the two enzymes. Obtaining structural information about complexes of LPMOs with carbohydrates has proven to be very difficult in general judging from the structures reported in the literature thus far, and this can be attributed only partly to the low affinity for small substrates. We have thus evaluated the use of differential scanning fluorimetry as a guide to obtaining complex structures. Furthermore, an analysis of crystal packing of LPMOs and glycoside hydrolases corroborates the hypothesis that active site occlusion is a very significant problem for LPMO-substrate interaction analysis by crystallography, due to their relatively flat and extended substrate binding sites.

AB - Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that cleave polysaccharide substrates oxidatively. First discovered because of their action on recalcitrant crystalline substrates (chitin and cellulose), a number of LPMOs are now reported to act on soluble substrates, including oligosaccharides. However, crystallographic complexes with oligosaccharides have been reported for only a single LPMO so far, an enzyme from the basidiomycete fungus Lentinus similis (LsAA9_A). Here we present a more detailed comparative study of LsAA9_A and an LPMO from the ascomycete fungus Collariella virescens (CvAA9_A) with which it shares 41.5% sequence identity. LsAA9_A is considerably more thermostable than CvAA9_A, and the structural basis for the difference has been investigated. We have compared the patterns of oligosaccharide cleavage and the patterns of binding in several new crystal structures explaining the basis for the product preferences of the two enzymes. Obtaining structural information about complexes of LPMOs with carbohydrates has proven to be very difficult in general judging from the structures reported in the literature thus far, and this can be attributed only partly to the low affinity for small substrates. We have thus evaluated the use of differential scanning fluorimetry as a guide to obtaining complex structures. Furthermore, an analysis of crystal packing of LPMOs and glycoside hydrolases corroborates the hypothesis that active site occlusion is a very significant problem for LPMO-substrate interaction analysis by crystallography, due to their relatively flat and extended substrate binding sites.

U2 - 10.1021/acs.biochem.0c00312

DO - 10.1021/acs.biochem.0c00312

M3 - Journal article

C2 - 32818374

AN - SCOPUS:85091125130

VL - 59

SP - 3347

EP - 3358

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 36

ER -

ID: 254669710