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Cloning, expression and characterization of xylose isomerase, XylA, from Caldanaerobacter subterraneus subsp. yonseiensis |
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| Authors: | Byoung-Chan Kim Sun Nyoung Yu Kwang Youn Kim Jung Sook Lee Yu-Ryang Pyun Soon Cheol Ahn |
| Institution: | 1. Biological Resource Center (BRC), Korean Collection for Type Cultures (KCTC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Yuseong-gu, Daejeon, 305-806, South Korea 2. Department of Microbiology and Immunology, Pusan National University School of Medicine, Yangsan, 626-870, Korea 3. Medical Research Institute, Pusan National University Hospital, Busan, 602-739, South Korea 4. Department of Biotechnology, Yonsei University, 134 Sinchon-dong, Seodaemun-gu, Seoul, 120-749, South Korea |
| Abstract: | The xylA gene, coding for xylose isomerase, from the extreme thermophile, Caldanaerobacter subterraneus subsp. yonseiensis was cloned, sequenced, and expressed in Escherichia coli. The nucleotide sequence of the xylA gene encoded a polypeptide of 438 residues with a calculated molecular weight of 50,170 Da. The purified XylA showed high sequence homology (92% identity) with that of Thermoanaerobacter thermohydrosulfuricus. The recombinant enzyme expressed in Escherichia coli was purified by heat treatment and gel chromatography. The purified enzyme was thermostable with optimal activity at 95°C. The enzyme required divalent cations including Zn2+ for its maximal activity and thermostability. |
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