Isolation and properties of a particulate fraction for the desaturation of palmitic acid from Alcaligenes faecalis.
A particulate fraction obtained from Alcaligenes faecalis could desaturate palmitic acid to palmitoleic acid. NADPH, ATP, CoA, Fe2+ and Mg2+ were essential cofactors for the reaction. The desaturation showed an absolute requirement for O2. Metal ions like Mn2+, Mo6+ and Cu2+ did not affect the desaturation, while Zn2+ was inhibitory. Sulfhydryl agents such as cysteine, glutathione and beta-mercaptoethanol had no effect, but SH-blocking agents like HgCl2 and p-hydroxymercuribenzoate inhibited the reaction. Azide and cyanide strongly inhibited the reaction while CO had no effect. The presence of a b-type cytochrome in the enzyme preparation was confirmed by the spectral studies on the reaction of enzyme with NADPH. Involvement of b-type cytochrome in the desaturation reaction was demonstrated by the reoxidation of b-type cytochrome initially reduced with NADPH, by the addition of palmitic acid and other cofactors. The pH optimum for the enzyme activity was 7.4. The optimum temperature for enzyme activity was 25 degrees C and maximum activity was obtained at the end of 45 min.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000421:Alcaligenes / Q000201:enzymology*; D002855:Chromatography, Thin Layer; D003065:Coenzyme A / Q000378:metabolism; D003580:Cytochromes / Q000378:metabolism; D044943:Fatty Acid Desaturases / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010169:Palmitic Acids / Q000378:metabolism*
D003580:Cytochromes; D010169:Palmitic Acids; D009249:NADP; D000255:Adenosine Triphosphate; D007501:Iron; D044943:Fatty Acid Desaturases; D008274:Magnesium; D003065:Coenzyme A