Lipid hydrogenation induces elevated 18:1-CoA desaturase activity in Candida lipolytica microsomes.
Microsomal membranes prepared from the mesophilic yeast Candida lipolytica grown at 10 degrees C were hydrogenated by the homogeneous Pd-catalyst, palladium di (sodium alizarine sulfonate) (Pd(QS)2). After hydrogenation to various levels, the microsomes were washed free of the Pd-complex and transferred to a reaction mixture (containing NADH, MgCl2, ATP, CoA and [14C]18:1-CoA) for assay of 18:1-CoA desaturase activity. Microviscosity alterations were also followed by measuring changes in DPH fluorescence polarization. Rapid catalytic hydrogenation of unsaturated fatty acids of the lipids occurred within 20-120 s, resulting in large increases in 16:0, 18:0 and 18:1 acids and decreases in 18:2 acid. In the range 7-20% 18:0 content, a pronounced increase in desaturase activity was observed, with a maximum of greater than 2-fold at a 18:0 content of 12%, followed by a decrease to the initial activity at 33% 18:0 content. These changes were well-correlated with changes in microviscosity, maximal desaturase activity occurring in the DPH fluorescence anisotropy range of 0.23-0.24; above and below this range, desaturase activities were close to the initial control values. It is suggested that the hydrogenation-induced increase in the formation of 18:2 from 18:1-CoA (proceeding partly through direct desaturation of PC) may be due to changes in conformation of the membrane-bound desaturase enzyme complex as a result of controlled rigidification of the surrounding lipids. The operation of such a self-regulating control mechanism would be consistent with a previously proposed model for microsomal desaturase action.
Institute of Biochemistry, BRC, Szeged, Hungary.
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Horváth
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Török
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Vígh
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Kates
D002175:Candida / Q000201:enzymology*; D044943:Fatty Acid Desaturases / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D005454:Fluorescence Polarization; D006865:Hydrogenation; D050356:Lipid Metabolism; D008861:Microsomes / Q000201:enzymology*; D013230:Stearoyl-CoA Desaturase / Q000378:metabolism*
D005227:Fatty Acids; D044943:Fatty Acid Desaturases; D013230:Stearoyl-CoA Desaturase