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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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.
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.
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.