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Structural features on the substrate-binding surface of fungal lytic polysaccharide monooxygenases determine their oxidative regioselectivity

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Abstract
Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that oxidatively cleave many of nature's most recalcitrant polysaccharides by acting on the C1- and/or C4-carbon of the glycosidic bond. Here, the results of an extensive mutagenesis study on three LPMO representatives, Phanerochaete chrysosporium LPMO9D (C1-oxidizer), Neurospora crassa LPMO9C (C4), and Hypocrea jecorina LPMO9A (C1/C4), are reported. Using a previously published indicator diagram, the authors demonstrate that several structural determinants of LPMOs play an important role in their oxidative regioselectivity. N-glycan removal and alterations of the aromatic residues on the substrate-binding surface are shown to alter C1/C4-oxidation ratios. Removing the carbohydrate binding module (CBM) is found not to alter the regioselectivity of HjLPMO9A, although the effect of mutational changes is shown to increase in a CBM-free context. The accessibility to the solvent-exposed axial position of the copper-site reveales not to be a major regioselectivity indicator, at least not in PcLPMO9D. Interestingly, a HjLPMO9A variant lacking two surface exposed aromatic residues combines decreased binding capacity with a 22% increase in synergetic efficiency. Similarly to recent LPMO10 findings, our results suggest a complex matrix of surface-interactions that enables LPMO9s not only to bind their substrate, but also to accurately direct their oxidative force.
Keywords
enzyme engineering, lytic polysaccharide monooxygenase, oxidative regioselectivity, Pichia pastoris, SERRATIA-MARCESCENS, CRYSTAL-STRUCTURE, CELLULOSE, DEGRADATION, CLEAVAGE, FAMILY, DIVERSITY, PROTEINS, INSIGHTS, REVEALS

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Chicago
Danneels, Barbara, Magali Tanghe, and Tom Desmet. 2019. “Structural Features on the Substrate-binding Surface of Fungal Lytic Polysaccharide Monooxygenases Determine Their Oxidative Regioselectivity.” Biotechnology Journal 14 (3).
APA
Danneels, B., Tanghe, M., & Desmet, T. (2019). Structural features on the substrate-binding surface of fungal lytic polysaccharide monooxygenases determine their oxidative regioselectivity. BIOTECHNOLOGY JOURNAL, 14(3).
Vancouver
1.
Danneels B, Tanghe M, Desmet T. Structural features on the substrate-binding surface of fungal lytic polysaccharide monooxygenases determine their oxidative regioselectivity. BIOTECHNOLOGY JOURNAL. 2019;14(3).
MLA
Danneels, Barbara, Magali Tanghe, and Tom Desmet. “Structural Features on the Substrate-binding Surface of Fungal Lytic Polysaccharide Monooxygenases Determine Their Oxidative Regioselectivity.” BIOTECHNOLOGY JOURNAL 14.3 (2019): n. pag. Print.
@article{8605440,
  abstract     = {Lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that oxidatively cleave many of nature's most recalcitrant polysaccharides by acting on the C1- and/or C4-carbon of the glycosidic bond. Here, the results of an extensive mutagenesis study on three LPMO representatives, Phanerochaete chrysosporium LPMO9D (C1-oxidizer), Neurospora crassa LPMO9C (C4), and Hypocrea jecorina LPMO9A (C1/C4), are reported. Using a previously published indicator diagram, the authors demonstrate that several structural determinants of LPMOs play an important role in their oxidative regioselectivity. N-glycan removal and alterations of the aromatic residues on the substrate-binding surface are shown to alter C1/C4-oxidation ratios. Removing the carbohydrate binding module (CBM) is found not to alter the regioselectivity of HjLPMO9A, although the effect of mutational changes is shown to increase in a CBM-free context. The accessibility to the solvent-exposed axial position of the copper-site reveales not to be a major regioselectivity indicator, at least not in PcLPMO9D. Interestingly, a HjLPMO9A variant lacking two surface exposed aromatic residues combines decreased binding capacity with a 22\% increase in synergetic efficiency. Similarly to recent LPMO10 findings, our results suggest a complex matrix of surface-interactions that enables LPMO9s not only to bind their substrate, but also to accurately direct their oxidative force.},
  articleno    = {1800211},
  author       = {Danneels, Barbara and Tanghe, Magali and Desmet, Tom},
  issn         = {1860-6768},
  journal      = {BIOTECHNOLOGY JOURNAL},
  language     = {eng},
  number       = {3},
  pages        = {10},
  title        = {Structural features on the substrate-binding surface of fungal lytic polysaccharide monooxygenases determine their oxidative regioselectivity},
  url          = {http://dx.doi.org/10.1002/biot.201800211},
  volume       = {14},
  year         = {2019},
}

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