PMID: 10739959Mar 31, 2000

Functional expression of two Arabidopsis aldehyde oxidases in the yeast Pichia pastoris

Journal of Biochemistry
Hanae KoiwaiT Koshiba

Abstract

To investigate the biochemical and enzymatic properties of two aldehyde oxidase (AO) isoforms of Arabidopsis thaliana, we expressed AAO1 and AAO2 cDNAs in a heterologous yeast (Pichia pastoris) system and successfully obtained the proteins in active forms. The expressed AAO1 and AAO2 proteins gave activity bands with the same mobilities on native gel electrophoresis and exhibited the same substrate preferences on zymograms with 8 aldehydes as those of AOalpha and AOgamma in Arabidopsis seedlings, respectively. Furthermore, anti-AAO1 and anti-AAO2 antibodies, which specifically recognize the seedling AOalpha and AOgamma, respectively, reacted with the AAO1 and AAO2 proteins produced in P. pastoris, respectively. These results indicate that these AO proteins are accurately produced in the yeast system, as in Arabidopsis seedlings. Using AO preparations from P. pastoris, the enzymatic properties of Arabidopsis AOalpha and AOgamma were investigated. AOalpha showed a relatively wide substrate specificity for 7 aldehydes tested, with high affinity to benzaldehyde and indole-3-aldehyde, while AOgamma could most efficiently oxidize naphthaldehyde. AOalpha was strongly inhibited by iodoacetate and KCN, while AOgamma was inhibited not on...Continue Reading

Citations

Oct 16, 2012·Plant Molecular Biology·Maryam ZarepourFlorian Bittner
Sep 6, 2000·The Plant Journal : for Cell and Molecular Biology·M SeoT Koshiba
Sep 29, 2012·The Journal of Biological Chemistry·Catarina CoelhoMaria João Romão
Apr 4, 2014·Chemical Reviews·Luisa B Maia, José J G Moura
Oct 6, 2018·Bioscience, Biotechnology, and Biochemistry·Mami TakeiYoshihito Suzuki
Apr 13, 2020·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Breeanna MintmierPartha Basu

Related Concepts

AAO1 protein, Arabidopsis
Aldehyde Oxidoreductases
Aldehydes
Enzyme Inhibitors
Hydrogen-Ion Concentration
Alloenzymes
Pichia
Precipitin Tests
Proteins, Recombinant DNA
Substrate Specificity

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