Analysis of the amino acid residues involved in the thermal properties of the monomeric isocitrate dehydrogenases of the psychrophilic bacterium Colwellia maris and the mesophilic bacterium Azotobacter vinelandii

Bioscience, Biotechnology, and Biochemistry
Takayuki Kurihara, Yasuhiro Takada

Abstract

Cold-adapted monomeric isocitrate dehydrogenase of a psychrophilic bacterium, Colwellia maris, (CmIDH) showed a high degree of amino acid sequential identity (69.5%) to a mesophilic nitrogen-fixing bacterium, Azotobacter vinelandii, (AvIDH). In this study, three Ala residues of CmIDH and the corresponding Pro residues of AvIDH were exchanged by site-directed mutagenesis, and several properties of single, double, and triple mutants of the two enzymes were investigated. The mutated CmIDHs, which replaced Ala719 with Pro, showed increased activity and elevation of the optimum temperature and thermostability for activity. In contrast, mutants of AvIDH, in which Pro717 was replaced by Ala, decreased the thermostability for activity. These results indicate that Ala719 of CmIDH and Pro717 of AvIDH are involved in thermostability. On the other hand, mutated CmIDH, in which Ala710 was replaced by Pro, and the corresponding AvIDH mutant, which replaced Pro708 with Ala, showed higher and lower specific activity than the corresponding wild-type enzymes, suggesting that Pro708 of AvIDH is involved in its high catalytic ability. Furthermore, the exchange mutations between Ala740 in CmIDH and the corresponding Pro738 in AvIDH resulted in decr...Continue Reading

References

Nov 26, 1998·International Journal of Systematic Bacteriology·I YumotoH Okuyama
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Jan 11, 2001·Biochimica Et Biophysica Acta·P P SheridanJ E Brenchley
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Jan 19, 2006·Extremophiles : Life Under Extreme Conditions·Shinsuke MakiYasuhiro Takada
Mar 11, 2010·Extremophiles : Life Under Extreme Conditions·Rami Al KhudaryGarabed Antranikian
Apr 8, 2011·Protein Engineering, Design & Selection : PEDS·Ronny MartinezDanilo Roccatano
Jan 1, 2006·CSH Protocols·Joseph Sambrook, David W Russell
Jan 1, 2006·CSH Protocols·Joseph Sambrook, David W Russell
Jan 1, 2006·CSH Protocols·Joseph Sambrook, David W Russell
Jan 1, 2006·CSH Protocols·Joseph Sambrook, David W Russell

Related Concepts

Azotobacter
Enzyme Stability
Alkalescens-Dispar Group
Isocitrate Dehydrogenase-I
Amino Acid Substitution
Alteromonadaceae
Biocatalysis
Alanine
Isocitrate Dehydrogenase (NAD+)
Nitrogen

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