Crystal structure of a family 6 cellobiohydrolase from the basidiomycete Phanerochaete chrysosporium

Mikako Tachioka, Akihiko Nakamura, Takuya Ishida, Kiyohiko Igarashi, Masahiro Samejima

    Research output: Contribution to journalArticleScientificpeer-review

    7 Citations (Scopus)

    Abstract

    Cellobiohydrolases belonging to glycoside hydrolase family 6 (CBH II, Cel6A) play key roles in the hydrolysis of crystalline cellulose. CBH II from the white-rot fungus Phanerochaete chrysosporium (PcCel6A) consists of a catalytic domain (CD) and a carbohydrate-binding module connected by a linker peptide, like other known fungal cellobiohydrolases. In the present study, the CD of PcCel6A was crystallized without ligands, and p-nitrophenyl ß-d-cellotrioside (pNPG3) was soaked into the crystals. The determined structures of the ligand-free and pNPG3-soaked crystals revealed that binding of cellobiose at substrate subsites +1 and +2 induces a conformational change of the N-terminal and C-terminal loops, switching the tunnel-shaped active site from the open to the closed form.The crystal structure of the catalytic domain of a glycoside hydrolase family 6 cellobiohydrolase from the basidiomycete P. chrysosporium was solved in apo and cellobiose-liganded forms at 1.2 and 2.1 Å resolution, respectively.
    Original languageEnglish
    Pages (from-to)398-403
    JournalActa Crystallographica Section F: Structural Biology Communications
    Volume73
    Issue number7
    DOIs
    Publication statusPublished - 2017
    MoE publication typeA1 Journal article-refereed

    Funding

    This research was supported by a Grant-in-Aid for Innovative Areas from the Japanese Ministry of Education, Culture, Sports and Technology (MEXT) to KI (Nos. 24114001 and 24114008), a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS) to KI (B, No. 24380089), an Impulsing Paradigm Change through Disruptive Technologies (ImPACT) from the Japan Science and Technology Agency (JST) to KI and a Grant-in-Aid for JSPS Fellows from JSPS to MT (15J10657).

    Keywords

    • biomass utilization
    • carbohydrate-active enzymes
    • cellobiohydrolase
    • cellulases
    • Phanerochaete chrysosporium
    • Trisaccharides/chemistry
    • Protein Conformation, alpha-Helical
    • Catalytic Domain
    • Gene Expression
    • Nitrobenzenes/chemistry
    • Phanerochaete/chemistry
    • Models, Molecular
    • Substrate Specificity
    • Cellulose 1,4-beta-Cellobiosidase/chemistry
    • Crystallography, X-Ray
    • Pichia/genetics
    • Amino Acid Motifs
    • Recombinant Proteins/chemistry
    • Protein Conformation, beta-Strand
    • Fungal Proteins/chemistry
    • Cloning, Molecular
    • Hydrophobic and Hydrophilic Interactions
    • Protein Binding
    • Protein Interaction Domains and Motifs

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