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Thermostable Hemicellulases of a Bacterium, Geobacillus sp. DC3, Isolated from the Former Homestake Gold Mine in Lead, South Dakota

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Abstract

A thermophilic strain, Geobacillus sp. DC3, capable of producing hemicellulolytic enzymes was isolated from the 1.5-km depth of the former Homestake gold mine in Lead, South Dakota. The DC3 strain expressed a high level of extracellular endoxylanase at 39.5 U/mg protein with additional hemicellulases including β-xylosidase (0.209 U/mg) and arabinofuranosidase (0.230 U/mg), after the bacterium was grown in xylan for 24 h. Partially purified DC3 endoxylanase exhibited a molecular mass of approximately 43 kDa according to zymography with an optimal pH of 7 and optimal temperature of 70 °C. The kinetic constants, K m and V max, were 13.8 mg/mL and 77.5 μmol xylose/min·mg xylan, respectively. The endoxylanase was highly stable and maintained 70 % of its original activity after 16 h incubation at 70 °C. The thermostable properties and presence of three different hemicellulases of Geobacillus sp. DC3 strain support its potential application for industrial hydrolysis of renewable biomass such as lignocelluloses.

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Acknowledgments

The authors wish to thank Dr. Cynthia Anderson and Dr. Dave Bergmann at Black Hills State University for providing valuable help in obtaining and interpreting sequencing data. Additional thanks go toward Dr. Todd Menkhaus for lending equipment for this project. Part of this work was supported by the South Dakota NASA EPSCoR Research Infrastructure Development (RID) Grant No. NNX07AL04A and South Dakota Board of Regents (SDBOR/BHSU 2011-19005) as a subaward from Black Hills State University (BHSU/SDSMT BA1100002). Additional support was received from the South Dakota 2010 Center of Bioprocessing Research and Development.

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  1. Department of Chemical and Biological Engineering, South Dakota School of Mines and Technology, 501 East Saint Joseph Street, Rapid City, SD, 57701, USA

    Terran E. Bergdale & Sookie S. Bang

  2. Renewable Product Technology Research Unit, National Center for Agricultural Utilization Research (NCAUR), Agricultural Research Service (ARS), United States Department of Agriculture (USDA), 1815 North University Street, Peoria, IL, 61604, USA

    Stephen R. Hughes

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Bergdale, T.E., Hughes, S.R. & Bang, S.S. Thermostable Hemicellulases of a Bacterium, Geobacillus sp. DC3, Isolated from the Former Homestake Gold Mine in Lead, South Dakota. Appl Biochem Biotechnol 172, 3488–3501 (2014). https://doi.org/10.1007/s12010-014-0784-7

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