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http://dx.doi.org/10.3839/jabc.2009.019

Cloning and Biochemical Characterization of Aspartate Aminotransferase from Xanthomonas oryzae pv. oryzae  

Kang, Han-Chul (Department of Functional Bio-material, National Academy of Agricultural Science, Rural Development Administration)
Yoon, Sang-Hong (Department of Functional Bio-material, National Academy of Agricultural Science, Rural Development Administration)
Lee, Chang-Mook (Department of Functional Bio-material, National Academy of Agricultural Science, Rural Development Administration)
Publication Information
Journal of Applied Biological Chemistry / v.52, no.3, 2009 , pp. 109-115 More about this Journal
Abstract
The gene encoding a putative aspartate aminotransferase in Xanthomonas oryzae pv. oryzae (Xoo) was cloned using PCR technique. The gene was ligated with pET-21(a) vector containing His6 tag and expressed in E. coli BL21(DE3). Affinity purification of the recombinant aspartate aminotransferase with Ni-NTA resin resulted in one band by SDS-PAGE analysis. The purified enzyme showed a molecular weight of 43 kDa, as expected. The enzyme was the most active toward L-aspartate as an amino donor, indicating that the purified enzyme is one of aspartate aminotrans-ferases exist in Xoo. Optimal activity of the enzyme was observed at around pH 7.5 and stability was much higher at alkaline pH rather than acidic pH values. The enzyme was considerably activated by the presence of manganese ion, showing about 157% of control activity at 1.0 mM.
Keywords
aspartate aminotransferase; Xanthomonas oryzae pv. oryzae;
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