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Int. J. Mol. Sci. 2014, 15(4), 5717-5729; doi:10.3390/ijms15045717

Cloning, Expression and Characterization of a Novel Thermophilic Polygalacturonase from Caldicellulosiruptor bescii DSM 6725

Key Laboratory for Molecular Enzymology & Engineering of the Ministry of Education, School of Life Science, Jilin University, Changchun 130012, China
School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China
These authors contributed equally to this work.
Author to whom correspondence should be addressed.
Received: 10 February 2014 / Revised: 25 March 2014 / Accepted: 26 March 2014 / Published: 3 April 2014
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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We cloned the gene ACM61449 from anaerobic, thermophilic Caldicellulosiruptor bescii, and expressed it in Escherichia coli origami (DE3). After purification through thermal treatment and Ni-NTA agarose column extraction, we characterized the properties of the recombinant protein (CbPelA). The optimal temperature and pH of the protein were 72 °C and 5.2, respectively. CbPelA demonstrated high thermal-stability, with a half-life of 14 h at 70 °C. CbPelA also showed very high activity for polygalacturonic acid (PGA), and released monogalacturonic acid as its sole product. The Vmax and Km of CbPelA were 384.6 U·mg−1 and 0.31 mg·mL−1, respectively. CbPelA was also able to hydrolyze methylated pectin (48% and 10% relative activity on 20%–34% and 85% methylated pectin, respectively). The high thermo-activity and methylated pectin hydrolization activity of CbPelA suggest that it has potential applications in the food and textile industry. View Full-Text
Keywords: thermophilic polygalacturonase; CbPelA; exo-PGase; Caldicellulosiruptor bescii thermophilic polygalacturonase; CbPelA; exo-PGase; Caldicellulosiruptor bescii

This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Chen, Y.; Sun, D.; Zhou, Y.; Liu, L.; Han, W.; Zheng, B.; Wang, Z.; Zhang, Z. Cloning, Expression and Characterization of a Novel Thermophilic Polygalacturonase from Caldicellulosiruptor bescii DSM 6725. Int. J. Mol. Sci. 2014, 15, 5717-5729.

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