Cloning,Expression and Characterization of a Novel Thermophilic Polygalacturonase from Caldicellulosiruptor bescii DSM 6725 |
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| Authors: | Yanyan Chen Dejun Sun Yulai Zhou Liping Liu Weiwei Han Baisong Zheng Zhi Wang Zuoming Zhang |
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| Affiliation: | 1.Key Laboratory for Molecular Enzymology & Engineering of the Ministry of Education, School of Life Science, Jilin University, Changchun 130012, China; E-Mails: (Y.C.); (L.L.); (W.H.); (B.Z.); (Z.W.);2.School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China; E-Mails: (D.S.); (Y.Z.) |
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| Abstract: | We cloned the gene {"type":"entrez-protein","attrs":{"text":"ACM61449","term_id":"222457187","term_text":"ACM61449"}}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. |
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| Keywords: | thermophilic polygalacturonase CbPelA exo-PGase Caldicellulosiruptor bescii |
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