Enhanced chemiluminescence: a sensitive analytical system for detection of sweet potato peroxidase.
3-(10'-Phenothiazinyl)propane-1-sulfonate (SPTZ) was shown to be a potent enhancer of anionic sweet potato peroxidase (aSPP)-induced chemiluminescence. The optimal conditions for aSPP-catalyzed oxidation of luminol were investigated by varying the concentrations of luminol, hydrogen peroxide, Tris, and SPTZ as well as the pH values of the reaction mixture. Addition of 4-morpholinopyridine (MORP) to the reaction mixture markedly increased the light intensity. Using SPTZ and MORP together enhanced the effect 265 times. The lower detection limit (LDL) of SPP was 0.09 pM, approximately in 10 times lower than that for the cationic isozyme c of horseradish peroxidase/4-iodophenol system. It was shown that aSPP in the presence of SPTZ produced a longer lasting chemiluminescent signal.
Vdovenko
Marina M
MM
Chemical Enzymology Department, M.V. Lomonosov Moscow State University, Moscow, Russia.
Della Ciana
Leopoldo
L
Sakharov
Ivan Y
IY
D017738:Alkanesulfonic Acids / Q000737:chemistry; D001709:Biotechnology / Q000379:methods*; D006861:Hydrogen Peroxide / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D027723:Ipomoea batatas / Q000201:enzymology*; D007700:Kinetics; D008163:Luminescent Measurements / Q000379:methods*; D008165:Luminol / Q000737:chemistry / Q000378:metabolism; D009025:Morpholines / Q000737:chemistry; D009195:Peroxidase / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013843:Thiazines / Q000737:chemistry; D014325:Tromethamine / Q000737:chemistry
C543492:3-(10'-Phenothiazinyl)propane-1-sulfonate; D017738:Alkanesulfonic Acids; D009025:Morpholines; D010940:Plant Proteins; D013843:Thiazines; D014325:Tromethamine; D008165:Luminol; D006861:Hydrogen Peroxide; D009195:Peroxidase