期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 111, 期 38, 页码 13822-13827出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1408090111
关键词
copper enzymes; oxygen activation; CBM20
资金
- National Institutes of Health [P30-EB009998]
- US Department of Energy [DE-AC02-98CH10886]
- Energy Biosciences Institute
- Scripps Research Institute
The recently discovered fungal and bacterial polysaccharide mono-oxygenases PMOs) are capable of oxidatively cleaving chitin, cellulose, and hemicelluloses that contain beta(1 -> 4) linkages between glucose or substituted glucose units. They are also known collectively as lytic PMOs, or LPMOs, and individually as AA9 formerly GH61), AA10 formerly CBM33), and AA11 enzymes. PMOs share several conserved features, including a monocopper center coordinated by a bidentate N-terminal histidine residue and another histidine ligand. A bioinformatic analysis using these conserved features suggested several potential new PMO families in the fungus Neurospora crassa that are likely to be active on novel substrates. Herein, we report on NCU08746 that contains a C-terminal starch-binding domain and an N-terminal domain of previously unknown function. Biochemical studies showedthatNCU08746requires copper, oxygen, and a source of electrons to oxidize the C1 position of glycosidic bonds in starch substrates, but not in cellulose or chitin. Starch contains alpha(1 -> 4) and a1. 6) linkages and exhibits higher order structures compared with chitin and cellulose. Cellobiose dehydrogenase, the biological redox partner of cellulose-active PMOs, can serve as the electron donor for NCU08746. NCU08746 contains one copper atom per protein molecule, which is likely coordinated by two histidine ligands as shown by X-ray absorption spectroscopy and sequence analysis. Results indicate that NCU08746 and homologs are starch-active PMOs, supporting the existence of a PMO superfamily with a much broader range of substrates. Starch-active PMOs provide an expanded perspective on studies of starch metabolism and may have potential in the food and starch-based biofuel industries.
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