Recombinant Cyclodextrinase from Thermococcus kodakarensis KOD1: Expression, Purification, and Enzymatic CharacterizationReport as inadecuate




Recombinant Cyclodextrinase from Thermococcus kodakarensis KOD1: Expression, Purification, and Enzymatic Characterization - Download this document for free, or read online. Document in PDF available to download.

Archaea - Volume 2015 2015, Article ID 397924, 8 pages -

Research Article

College of Plant Sciences, Jilin University, Changchun 130062, China

Department of Life Science, Chung-Ang University, Seoul 156-756, Republic of Korea

Received 5 September 2014; Revised 15 December 2014; Accepted 7 January 2015

Academic Editor: María J. Bonete

Copyright © 2015 Ying Sun et al. 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

A gene encoding a cyclodextrinase from Thermococcus kodakarensis KOD1 CDase-Tk was identified and characterized. The gene encodes a protein of 656 amino acid residues with a molecular mass of 76.4 kDa harboring four conserved regions found in all members of the α-amylase family. A recombinant form of the enzyme was purified by ion-exchange chromatography, and its catalytic properties were examined. The enzyme was active in a broad range of pH conditions pHs 4.0–10.0, with an optimal pH of 7.5 and a temperature optimum of 65°C. The purified enzyme preferred to hydrolyze β-cyclodextrin CD but not α- or γ-CD, soluble starch, or pullulan. The final product from β-CD was glucose. The and values were 3.13 ± 0.47 U mg

and 2.94 ± 0.16 mg mL

for β-CD. The unique characteristics of CDase-Tk with a low catalytic temperature and substrate specificity are discussed, and the starch utilization pathway in a broad range of temperatures is also proposed.





Author: Ying Sun, Xiaomin Lv, Zhengqun Li, Jiaqiang Wang, Baolei Jia, and Jinliang Liu

Source: https://www.hindawi.com/



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