Functional screening of hydrolytic activities reveals an extremely thermostable cellulase from a deep-sea archaeon

Benedikt Leis, Simon Heinze, Angel Angelov, Vu Thuy Trang Pham, Andrea Thürmer, Mohamed Jebbar, Peter N. Golyshin, Wolfgang R. Streit, Rolf Daniel, Wolfgang Liebl*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    24 Scopus citations


    Extreme habitats serve as a source of enzymes that are active under extreme conditions and are candidates for industrial applications. In this work, six large-insert mixed genomic libraries were screened for hydrolase activities in a broad temperature range (8-70°C). Among a variety of hydrolytic activities, one fosmid clone, derived from a library of pooled isolates of hyperthermophilic archaea from deep sea vents, displayed hydrolytic activity on carboxymethyl cellulose substrate plates at 70°C but not at lower temperatures. Sequence analysis of the fosmid insert revealed a gene encoding a novel glycoside hydrolase family 12 (GHF12) endo-1,4-β-glucanase, termed Cel12E. The enzyme shares 45% sequence identity with a protein from the archaeon Thermococcus sp. AM4 and displays a unique multidomain architecture. Biochemical characterization of Cel12E revealed a remarkably thermostable protein, which appears to be of archaeal origin. The enzyme displayed maximum activity at 92°C and was active on a variety of linear 1,4-β-glucans like carboxymethyl cellulose, β-glucan, lichenan, and phosphoric acid swollen cellulose. The protein is able to bind to various insoluble β-glucans. Product pattern analysis indicated that Cel12E is an endo-cleaving β-glucanase. Cel12E expands the toolbox of hyperthermostable archaeal cellulases with biotechnological potential.

    Original languageEnglish (US)
    Article number00095
    JournalFrontiers in Bioengineering and Biotechnology
    Issue numberJUL
    StatePublished - 2015


    • Archaeal endoglucanase
    • Enzymatic characterization
    • Extreme thermostable protein
    • Functional screenings

    ASJC Scopus subject areas

    • Biotechnology
    • Bioengineering
    • Histology
    • Biomedical Engineering


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