Contribution of Three Different Regions of Isocitrate Dehydrogenases from Psychrophilic and Psychrotolerant Bacteria to Their Thermal Properties

Current Microbiology
Yuka Mouri, Yasuhiro Takada

Abstract

Monomeric isocitrate dehydrogenases of a psychrophilic bacterium, Colwellia maris, and a psychrotolerant bacterium, Pseudomonas psychrophila, (CmIDH and PpIDH) are cold-adapted and mesophilic, respectively. On the other hand, previous studies revealed that the monomeric IDH of Azotobacter vinelandii (AvIDH) is also mesophilic and the regions 2 and 3 among three regions of this enzyme are involved in the thermal properties. Therefore, to examine whether the region(s) responsible for the mesophilic properties are common between PpIDH and AvIDH, the genes of chimeric IDHs exchanging three regions of PpIDH and CmIDH in various combinations were constructed and overexpressed as His-tagged recombinant proteins in the Escherichia coli cells, and the chimeric and wild-type PpIDH and CmIDH were purified with Ni-chelating affinity column chromatography. The swapping chimeras of the regions 2 or 3 in PpIDH and CmIDH showed lower and higher optimum temperatures for activities and their thermostabilities than the wild-type ones, respectively. On the other hand, the exchange of the respective region 1 hardly influenced these properties of the two IDHs. Therefore, the regions 2 and 3 of the two IDHs were confirmed to be involved in their ther...Continue Reading

References

Jun 7, 1974·Biochimica Et Biophysica Acta·W F BurkeH C Reeves
Aug 1, 1972·Journal of Bacteriology·J Y D'Aoust, D J Kushner
Nov 26, 1998·International Journal of Systematic Bacteriology·I YumotoH Okuyama
Jan 19, 2006·Extremophiles : Life Under Extreme Conditions·Shinsuke MakiYasuhiro Takada
Jun 16, 2010·Archives of Microbiology·Shougo MatsuoYasuhiro Takada

Methods Mentioned

BETA
column chromatography
PCR

Related Concepts

Protein Domain
Bacterial Proteins
Enzyme Stability
Isocitrate Dehydrogenase-I
Alteromonadaceae
Chelating Agents
Escherichia coli
Genes
Isocitrate Dehydrogenase (NAD+)
Recombinant Proteins

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