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Crystallization of a fungal lytic polysaccharide monooxygenase expressed from glycoengineered Pichia pastoris for X‐ray and neutron diffraction
Author(s) -
O'Dell William B.,
Swartz Paul D.,
Weiss Kevin L.,
Meilleur Flora
Publication year - 2017
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x16020318
Subject(s) - pichia pastoris , lytic cycle , crystallization , microbiology and biotechnology , chemistry , biology , biochemistry , recombinant dna , genetics , organic chemistry , gene , virus
Lytic polysaccharide monooxygenases (LPMOs) are carbohydrate‐disrupting enzymes secreted by bacteria and fungi that break glycosidic bonds via an oxidative mechanism. Fungal LPMOs typically act on cellulose and can enhance the efficiency of cellulose‐hydrolyzing enzymes that release soluble sugars for bioethanol production or other industrial uses. The enzyme PMO‐2 from Neurospora crassa ( Nc PMO‐2) was heterologously expressed in Pichia pastoris to facilitate crystallographic studies of the fungal LPMO mechanism. Diffraction resolution and crystal morphology were improved by expressing Nc PMO‐2 from a glycoengineered strain of P. pastoris and by the use of crystal seeding methods, respectively. These improvements resulted in high‐resolution (1.20 Å) X‐ray diffraction data collection at 100 K and the production of a large Nc PMO‐2 crystal suitable for room‐temperature neutron diffraction data collection to 2.12 Å resolution.