Expression of two old yellow enzyme homologues from Gluconobacter oxydans and identification of their citral hydrogenation abilities.
Two genes that encode proteins which share 30-35% sequence identity with yeast OYE (Old Yellow Enzyme, an NAD(P)H FMN-oxidoreductase), the well-studied archetype of the OYE protein family, have been identified in Gluconobacter oxydans M5. The two genes are localized in the chromosome and plasmid, respectively. Comparison of the deduced amino acid sequences of the enzymes with database entries revealed 75.1% similarity and 64.9% identity to that of the Pseudomonas syringae pv. glycinea NAD(P)H-dependent 2-cyclohexen-1-one reductase. The two proteins were expressed as His-tag fusion proteins in Escherichia coli and purified. The ability of the purified proteins to hydrogenate citral was identified. The results showed that the alpha,beta-double bond of citral cis-isomer 'neral' could be stereoselectively reduced to produce citronellal by the purified OYE homologues.
Yin
Bo
B
State key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.
Yang
Xuepeng
X
Wei
Guodong
G
Ma
Yushu
Y
Wei
Dongzhi
D
D000080462:Acyclic Monoterpenes; D000447:Aldehydes / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008401:Gas Chromatography-Mass Spectrometry; D020600:Gluconobacter oxydans / Q000201:enzymology* / Q000235:genetics; D006865:Hydrogenation; D015394:Molecular Structure; D039821:Monoterpenes / Q000737:chemistry / Q000378:metabolism*; D009252:NADPH Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D000080462:Acyclic Monoterpenes; D000447:Aldehydes; D039821:Monoterpenes; D011994:Recombinant Proteins; D009252:NADPH Dehydrogenase; C108217:citronellal; C007076:citral