Abstract
Caldicellulosiruptor bescii, the most thermophilic cellulolytic bacterium, is rich in hydrolytic and accessory enzymes that can degrade untreated biomass, but the precise role of many these enzymes is unknown. One of such enzymes is a predicted GDSL lipase or esterase encoded by the locus Athe_0553. In this study, this probable esterase named as Cbes-AcXE2 was overexpressed in Escherichia coli. The Ni–NTA affinity purified enzyme exhibited an optimum pH of 7.5 at an optimum temperature of 70 °C. Cbes-AcXE2 hydrolyzed p-nitrophenyl (pNP) acetate, pNP-butyrate, and phenyl acetate with approximately equal efficiency. The specific activity and K M for the most preferred substrate, phenyl acetate, were 142 U/mg and 0.85 mM, respectively. Cbes-AcXE2 removed the acetyl group of xylobiose hexaacetate and glucose pentaacetate like an acetyl xylan esterase (AcXE). Bioinformatics analyses suggested that Cbes-AcXE2, which carries an SGNH hydrolase-type esterase domain, is a member of an unclassified carbohydrate esterase (CE) family. Moreover, Cbes-AcXE2 is evolutionarily and biochemically similar to an unclassified AcXE, Axe2, of Geobacillus stearothermophilus. Thus, we proposed a novel family of carbohydrate esterase for both Cbes-AcXE2 and Axe2.
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Acknowledgements
We thank Prof. M.W.W Adams (University of Georgia, USA) for the kind gift of the genomic DNA of C. bescii. We also thank Dr. Ajit Tiwari for carefully reading the manuscript and providing his suggestions. This research was financially supported by Department of Biotechnology (Government of India) under the program Energy Bioscience Overseas Fellowship (No. BT/NBDB/22/06/2011) and DBT-ICT Centre for Energy Biosciences – Phase II (BT/EB/ICT-Extension/2012).
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Communicated by H. Atomi.
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Soni, S., Sathe, S.S., Odaneth, A.A. et al. SGNH hydrolase-type esterase domain containing Cbes-AcXE2: a novel and thermostable acetyl xylan esterase from Caldicellulosiruptor bescii . Extremophiles 21, 687–697 (2017). https://doi.org/10.1007/s00792-017-0934-2
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DOI: https://doi.org/10.1007/s00792-017-0934-2