Purification and characterization of recombinant acetohydroxyacid synthase from Heamophillus influazae
Author affiliations
DOI:
https://doi.org/10.15625/0866-7160/v38n3.8382Keywords:
Haemophillus influenzae, acetohydroxyacid synthase, enzymatic activity, inhibitor, purification.Abstract
Acetohydroxyacid synthase (AHAS) presents only in plants and microorganisms. The enzyme catalyzes the first common step in the biosynthesis of branch chain amino acids (BCAAs), including isoleucine, leucine and valine. AHAS is also a potential target for controlling Haemophilus influenzae. In this study, the recombinant catalytic subunit of AHAS from H. influenza (Hin-AHAS) was expressed in Escherichia coli. The purified Hin-AHAS protein exhibited a molecular weight of approximately 63 kDa on SDS-PAGE gel. The apparent Vmax and Km values of the purified Hin-AHAS were determined to be 0.236 U/mg protein and 2.503 mM pyruvate, respectively. Two inhibitors of plant AHAS, namely ethoxysulfuron (ETS) and pyrazosulfuron ethyl, were shown to inhibit Hin-AHAS in a non-competitive manner with the IC50 values of 90.14 µM and 376.6 µM, respectively. This result showed that the purified enzyme can be used for screening of inhibitors against Hin-AHAS.
Downloads
Downloads
Published
How to Cite
Issue
Section
License
Academia Journal of Biology (AJB) is an open-access and peer-reviewed journal. The articles published in the AJB are licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND 4.0), which permits for immediate free access to the articles to read, download, copy, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited (with a link to the formal publication through the relevant DOI), and without subscription charges or registration barriers. The full details of the CC BY-NC-ND 4.0 License are available at https://creativecommons.org/licenses/by-nc-nd/4.0/.
