Lipoxygenase from banana leaf: purification and characterization of an enzyme that catalyzes linoleic acid oxygenation at the 9-position.
The objective of the present study was to purify and characterize the lipoxygenase (LOX) from banana leaf (Giant Cavendishii, AAA), an unutilized bioresource. LOX was extracted, isolated, and purified 327-fold using 25-50% saturation of ammonium sulfate fractionation, hydroxyapatite column separation, and gel filtration on Superdex 200. The molecular mass of the purified LOX was 85 kDa, K(m) was 0.15 mM, and V(max) was 2.4 microM/min.mg using linoleic acid as substrate. Triton X-100 was required in the extraction medium; otherwise, no LOX activity was detected. LOX activity increased with the concentration of Triton X-100 with an optimum at 0.1%. The optimal pH of the purified LOX from banana leaf was 6.2, and optimal temperature was 40 degrees C. The LOX showed the highest reactivity toward 18:2 followed by 18:3 and 20:4. A very low reaction rate was observed toward 20:5 and 22:6. On the basis of retention time in normal phase HPLC, the products of 18:2 or 18:3 catalyzed by purified LOX were hydroperoxyoctadecadienoic acid or hydroperoxyoctadecatrienoic acid. It seems that 9-LOX is the predominant enzyme in banana leaf. Banada leaf dried at 110 degrees C for 2 h developed algal aroma. Banana leaf extract stored at 10 degrees C for 12 h formed an oolong tea-like flavor. Banana leaf extract reacted with 18:2 or soybean oil pretreated with bacterial lipase produced green and melon-like aroma, whereas the same reaction with 18:3 produced a sweet, fruity, cucumber-like flavor note.
Kuo
Jen-Min
JM
Department of Seafood Science, National Kaohsiung Marine University, Kaohsiung, Taiwan, R.O.C. ikuojm@mail.nkmu.edu.tw
Hwang
Ann
A
Yeh
Dong-Bor
DB
Pan
Min-Hsiung
MH
Tsai
Mei-Ling
ML
Pan
Bonnie Sun
BS
D001665:Binding Sites; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D019787:Linoleic Acid / Q000737:chemistry* / Q000378:metabolism*; D008084:Lipoxygenase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D028521:Musa / Q000201:enzymology*; D017830:Octoxynol; D009812:Odorants / Q000032:analysis; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000201:enzymology*
D010936:Plant Extracts; D017830:Octoxynol; D019787:Linoleic Acid; D008084:Lipoxygenase