Recombinant lipoxygenases as biocatalysts for natural flavour production.
We identified a lipoxygenase expressed early during almond seed development. Biochemical and molecular characterisation showed that the enzyme produces almost exclusively 9-hydroperoxides which have been demonstrated to be important factors for the production of characteristic aromas in several fruits. An almond LOX cDNA was identified by RT-PCR using RNA extracted from immature almond seeds. Sequence analysis revealed that the identified gene is closely related to tomato fruit and potato tuber lipoxygenases. The isolated cDNA was cloned into pET24a and the expression of recombinant protein was induced in E. coli. The presence of an active LOX was confirmed in cells containing the recombinant vector. HPLC analysis of the reaction products of recombinant almond LOX confirmed that the isolated cDNA encodes a 9-LOX.
Gallo
A
A
Istituto di Ricerca sulle Biotecnologie Agroalimentan, C.N.R., Via Prov. le Lecce-Monteroni, 73100 Lecee, Italy.
Mita
G
G
Zasiura
C
C
Casey
R
R
Santino
A
A
Zacheo
G
G
D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D031211:Corylus / Q000201:enzymology; D019649:Food Industry; D007700:Kinetics; D008084:Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D027861:Prunus / Q000201:enzymology / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000201:enzymology; D013649:Taste
D011994:Recombinant Proteins; D012313:RNA; D008084:Lipoxygenase