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

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

Received: 10 February 2014; in revised form: 25 March 2014 / Accepted: 26 March 2014 / Published: 3 April 2014
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: 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.
Keywords: thermophilic polygalacturonase; CbPelA; exo-PGase; Caldicellulosiruptor bescii
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MDPI and ACS Style

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.

AMA Style

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. International Journal of Molecular Sciences. 2014; 15(4):5717-5729.

Chicago/Turabian Style

Chen, Yanyan; Sun, Dejun; Zhou, Yulai; Liu, Liping; Han, Weiwei; Zheng, Baisong; Wang, Zhi; Zhang, Zuoming. 2014. "Cloning, Expression and Characterization of a Novel Thermophilic Polygalacturonase from Caldicellulosiruptor bescii DSM 6725." Int. J. Mol. Sci. 15, no. 4: 5717-5729.

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