PMID: 2502399Jul 15, 1989Paper

Stability and activity of a thermostable malic enzyme in denaturants and water-miscible organic solvents

European Journal of Biochemistry
A GuagliardiS Bartolucci

Abstract

A study was made of the effects of common protein denaturants and water-miscible organic solvents on both the stability and activity of the malic enzyme [(S)-malate:NADP+ oxidoreductase (oxaloacetate-decarboxylating); EC 1.1.1.40] from the extreme thermoacidophilic archaebacterium Sulfolobus solfataricus. At 25 degrees C, the enzyme was not inactivated in 4 M urea or 0.05% SDS over 24 h, while the half-life was 30 min in 6 M guanidine hydrochloride and 5 h in 0.075% SDS. The enzyme stability in water-miscible organic solvents at 25 degrees C is somewhat surprising: after a 24-h incubation, the enzyme was completely active in 50% dimethylformamide; it lost 15% of its initial activity in 50% methanol or 15% ethanol. However, the resistance to organic solvents was greatly reduced at higher temperatures. The enzyme was able to catalyze the malate conversion even in the presence of 1.5% Triton X-100 or sodium deoxycholate. A number of solvents were found to stimulate the malic activity independent of time. Studies with 50% methanol revealed that the activation was reversible and inversely related to the temperature; moreover, the solvent was demonstrated to exclusively affect the maximal velocity of catalysis, the Km values for both...Continue Reading

References

May 14, 1986·Biochemical and Biophysical Research Communications·B Norling
Dec 1, 1987·Protein Engineering·M Vihinen
Dec 1, 1982·The Biochemical Journal·R M DanielM P Curran
Dec 1, 1980·Analytical Biochemistry·A J Furth
Aug 13, 1981·Biochimica Et Biophysica Acta·R Lamed, J G Zeikus
Jul 1, 1987·Biotechnology and Bioengineering·C LaaneC Veeger

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Citations

Apr 23, 2008·Applied Microbiology and Biotechnology·Ritu Sareen, Prashant Mishra
Dec 25, 2010·Extremophiles : Life Under Extreme Conditions·Hasan Cihad Tekedar, Gülşah Sanlı-Mohamed
Nov 1, 1992·Enzyme and Microbial Technology·G CartaM E Gibson
Feb 1, 1996·The International Journal of Biochemistry & Cell Biology·A GuagliardiS Bartolucci
Apr 23, 2013·Journal of Structural Biology·Sabrina ReichBernhard Hauer
Jan 19, 2010·Seikagaku. The Journal of Japanese Biochemical Society·Hiroyasu Ogino
Nov 28, 2009·Chembiochem : a European Journal of Chemical Biology·Björn V AdalbjörnssonNigel S Scrutton
Sep 24, 1992·Biochimica Et Biophysica Acta·G RaimoV Bocchini

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