Isovaleryl CoA dehydrogenase (IVDH) is a flavoprotein that introduce a trans-double bond between C2 and C3 of the isovaleryl CoA substrate, an intermediate in leucine catabolism pathway. Interrogation of the Paracoccus denitrificans Pd1222 genome has identified Pden_3633 gene as a candidate to encode for IVDH. In previous study by Rafid et al (under the publishing), this putative IVDH was expressed in E. coli and purified as N-terminal Strep-tagged protein. In current study, spectral properties of the purified IVDH were conducted and the results showed that the enzyme was obtained as an apoprotein. For this reason, IVDH has been reconstituted by incubation with flavin adenine dinucleotide (FAD) and the experiment showed that the ratio 1: 20% (IVDH: FAD molar excess) was achieved fully reconstitution. Furthermore, kinetic parameters of the reconstituted IVDH were studied and the enzyme exhibited a specific activity for isovaleryl-CoA substrate while Vmax and KM were estimated to be 600 mU mg-1 and 7.13 μM respectively. Finally, gel filtration by using HiLoad 16/60 Superdex 200 column was performed to investigate the quaternary structure of the recombinant IVDH. An apparent molecular weight of 171640 Da was determined.
Spectral Properties, Reconstitution and Kinetics of Paracoccous denitrificans Isovaleryl-CoA Dehydrogenase
Acyl-CoA Dehydrogenase
Paracoccus denitrificans
Reconstitution
Publication Date
Tue Jan 18 2022
Journal Name
Materials Science Forum
The Effect of Gamma Radiation on the Manufactured HgBa<sub>2</sub>Ca<sub>2</sub>Cu<sub>2.4</sub>Ag<sub>0.6</sub>O<sub>8+δ</sub> Compound
Gamma doses radiation
crystal size
strain
degree of crystallinity
Scherrer
crystallinity and Williamson equations.
In this article four samples of HgBa2Ca2Cu2.4Ag0.6O8+δ were prepared and irradiated with different doses of gamma radiation 6, 8 and 10 Mrad. The effects of gamma irradiation on structure of HgBa2Ca2Cu2.4Ag0.6O8+δ samples were characterized using X-ray diffraction. It was concluded that there effect on structure by gamma irradiation. Scherrer, crystallization, and Williamson equations were applied based on the X-ray diffraction diagram and for all gamma doses, to calculate crystal size, strain, and degree of crystallinity. I
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