Applied Microbiology and Biotechnology

, Volume 100, Issue 4, pp 1823–1831 | Cite as

Deletion of a gene cluster for [Ni-Fe] hydrogenase maturation in the anaerobic hyperthermophilic bacterium Caldicellulosiruptor bescii identifies its role in hydrogen metabolism

  • Minseok Cha
  • Daehwan Chung
  • Janet Westpheling
Applied genetics and molecular biotechnology


The anaerobic, hyperthermophlic, cellulolytic bacterium Caldicellulosiruptor bescii grows optimally at ∼80 °C and effectively degrades plant biomass without conventional pretreatment. It utilizes a variety of carbohydrate carbon sources, including both C5 and C6 sugars, released from plant biomass and produces lactate, acetate, CO2, and H2 as primary fermentation products. The C. bescii genome encodes two hydrogenases, a bifurcating [Fe-Fe] hydrogenase and a [Ni-Fe] hydrogenase. The [Ni-Fe] hydrogenase is the most widely distributed in nature and is predicted to catalyze hydrogen production and to pump protons across the cellular membrane creating proton motive force. Hydrogenases are the key enzymes in hydrogen metabolism and their crystal structure reveals complexity in the organization of their prosthetic groups suggesting extensive maturation of the primary protein. Here, we report the deletion of a cluster of genes, hypABFCDE, required for maturation of the [Ni-Fe] hydrogenase. These proteins are specific for the hydrogenases they modify and are required for hydrogenase activity. The deletion strain grew more slowly than the wild type or the parent strain and produced slightly less hydrogen overall, but more hydrogen per mole of cellobiose. Acetate yield per mole of cellobiose was increased ∼67 % and ethanol yield per mole of cellobiose was decreased ∼39 %. These data suggest that the primary role of the [Ni-Fe] hydrogenase is to generate a proton gradient in the membrane driving ATP synthesis and is not the primary enzyme for hydrogen catalysis. In its absence, ATP is generated from increased acetate production resulting in more hydrogen produced per mole of cellobiose.


Anaerobe Hyperthermophile Caldicellulosiruptor bescii Hydrogen Bifurcating [Fe-Fe] hydrogenase [Ni-Fe] hydrogenase Hydrogenase maturation proteins 



We thank Jennifer Copeland and Elise Snyder for the outstanding technical assistance, Brian Davison for providing the switchgrass used in this study, Sidney Kushner for the expert technical advice, William Whitman for the advice and use of his GC, Joe Groom and Jenna Young for the critical review of the manuscript. The BioEnergy Science Center is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

253_2015_7025_MOESM1_ESM.pdf (356 kb)
ESM 1 (PDF 356 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Minseok Cha
    • 1
    • 2
  • Daehwan Chung
    • 1
    • 2
  • Janet Westpheling
    • 1
    • 2
  1. 1.Department of GeneticsUniversity of GeorgiaAthensUSA
  2. 2.The BioEnergy Science CenterOak Ridge National LaboratoryOak RidgeUSA

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