American Journal of Medical and Biological Research
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American Journal of Medical and Biological Research. 2016, 4(3), 42-52
DOI: 10.12691/ajmbr-4-3-2
Open AccessArticle

Role of Some Metal Ions on Steady–state Kinetics of Engineered Wild–type and Manganese (II) Binding Site Mutants of Recombinant Phlebia radiata Manganese Peroxidase 3 (rPr-MnP3)

Usenobong F. Ufot1, , Aniefiok E. Ite2, 3, Idorenyin H. Usoh1 and Monday I. Akpanabiatu4

1Department of Biological Sciences, Akwa Ibom State University, P.M.B. 1167, Uyo, Akwa Ibom State, Nigeria

2Department of Chemistry, Akwa Ibom State University, P.M.B. 1167, Uyo, Akwa Ibom State, Nigeria

3Research and Development Unit, Akwa Ibom State University, P.M.B. 1167, Uyo, Akwa Ibom State, Nigeria

4Department of Biochemistry, University of Uyo, P. M. B. 1017, Uyo, Akwa Ibom State, Nigeria

Pub. Date: August 06, 2016

Cite this paper:
Usenobong F. Ufot, Aniefiok E. Ite, Idorenyin H. Usoh and Monday I. Akpanabiatu. Role of Some Metal Ions on Steady–state Kinetics of Engineered Wild–type and Manganese (II) Binding Site Mutants of Recombinant Phlebia radiata Manganese Peroxidase 3 (rPr-MnP3). American Journal of Medical and Biological Research. 2016; 4(3):42-52. doi: 10.12691/ajmbr-4-3-2

Abstract

This study investigated the steady-state kinetics of engineered wild-type and manganese (II) binding site mutants of recombinant Phlebia radiata manganese peroxidase 3(rPr-MnP3). The effect (activation or inhibition) of some metal ions (Co2+, Zn2+ Cu2+ and Na+) on the activity of rPr-MnP3 enzymes was also studied. The results obtained showed that the rPr-MnP3 mutants in which the metal binding functionality has been largely lost have been created. Na+ (mono-valent ion) and Co2+showed similar characteristics by exhibiting stimulatory effects on the activity of wild-type rPr-MnP3. However, Cu2+ and Zn2+ had mixed inhibitory effects on wild-type and mutants (E40H, E44H, E40H/E44H). It was observed that Cu2+ was by far the strongest inhibitor of engineered rPr-MnP3 enzymes while Co2+ exhibited a non-competitive inhibitory effect on the double mutant (E40H/E44H) and D186H activities. In addition, Zn2+ and Cu2+also had non-competitive inhibitory effect on D186H mutant enzyme activity. The results obtained further showed that the competitive inhibitory effect of Cu2+observed in other rPr-MnP3 enzymes is largely removed in D186H mutant enzyme. Generally, histidine substitution retained a strong selectivity for Cu2+ as competitive inhibitor. Zn2+ being generally non-competitive suggest involvement of sites other than the Mn (II) binding site. This study showed that rPr-MnP3 enzymes function with alternate ligands in the Mn2+ binding site and does not have absolute obligate requirement for all carboxylate ligand set.

Keywords:
peroxidase Phlebiaradiata steady-state wild-type mutants metal ions inhibitors

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