Enzymatic carotenoid cleavage in star fruit (Averrhoa carambola).
This paper presents the first description of an enzyme fraction exhibiting carotenoid cleavage activity isolated from fruit skin of Averrhoa carambola. Partial purification of the enzyme could be achieved by acetone precipitation, ultrafiltration (300 kDa, 50 kDa), isoelectric focusing (pH 3-10) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (7.5%). In this way, an enzymatically active protein fraction was obtained, consisting of four proteins in the molecular weight range of between 12 and 90 kDa. Using beta-carotene as substrate, the enzyme activity was detected spectrophotometrically at 505 nm. The main reaction product, detected by GC analysis, was beta-ionone. This proves that the isolated enzymes are closely related to aroma metabolism and release of star fruit. The time constant of the reaction was 16.6 min, the Michaelis Constant K(m)=3.6 micromol 1(-1) and the maximum velocity V(max)=10.5 x 10(-3) micromol l(-1) s(-1) mg((Protein))(-1). The optimum temperature was 45 degrees C.
Fleischmann
Peter
P
Institut für Lebensmittelchemie, Technische Universität Braunschweig, Schleinitzstrasse 20, D-38106 Braunschweig, Germany. p.fleischmann@tu-bs.de
Watanabe
Naoharu
N
Winterhalter
Peter
P
D002338:Carotenoids / Q000737:chemistry / Q000378:metabolism*; D002849:Chromatography, Gas; D004591:Electrophoresis, Polyacrylamide Gel; D005638:Fruit / Q000201:enzymology*; D007525:Isoelectric Focusing; D007700:Kinetics; D045792:Norisoprenoids; D010084:Oxidation-Reduction; D027824:Rosaceae / Q000737:chemistry / Q000201:enzymology*; D013053:Spectrophotometry; D013696:Temperature; D013729:Terpenes / Q000378:metabolism; D019207:beta Carotene / Q000378:metabolism
D045792:Norisoprenoids; D013729:Terpenes; D019207:beta Carotene; D002338:Carotenoids; C008157:beta-ionone