Heterologous expression of Gaeumannomyces graminis lipoxygenase in Aspergillus nidulans.
Aspergillus sp. contain ppo genes coding for Ppo enzymes that produce oxylipins from polyunsaturated fatty acids. These oxylipins function as signal molecules in sporulation and influence the asexual to sexual ratio of Aspergillus sp. Fungi like Aspergillus nidulans and Aspergillus niger contain just ppo genes where the human pathogenic Aspergillus flavus and Aspergillus fumigatus contain ppo genes as well as lipoxygenases. Lipoxygenases catalyze the synthesis of oxylipins and are hypothesized to be involved in quorum-sensing abilities and invading plant tissue. In this study we used A. nidulans WG505 as an expression host to heterologously express Gaeumannomyces graminis lipoxygenase. The presence of the recombinant LOX induced phenotypic changes in A. nidulans transformants. Also, a proteomic analysis of an A. nidulans LOX producing strain indicated that the heterologous protein was degraded before its glycosylation in the secretory pathway. We observed that the presence of LOX induced the specific production of aminopeptidase Y that possibly degrades the G. graminis lipoxygenase intercellularly. Also the presence of the protein thioredoxin reductase suggests that the G. graminis lipoxygenase is actively repressed in A. nidulans.
Laboratory of Systems and Synthetic Biology, Microbial Systems and Synthetic Biology, Dreijenplein 10, Wageningen 6703 HB, Netherlands.
Ruud
R
Heshof
Laboratory of Systems and Synthetic Biology, Microbial Systems and Synthetic Biology, Dreijenplein 10, Wageningen 6703 HB, Netherlands.
J Paul
JP
van Schayck
Laboratory of Systems and Synthetic Biology, Microbial Systems and Synthetic Biology, Dreijenplein 10, Wageningen 6703 HB, Netherlands.
Juan Antonio
JA
Tamayo-Ramos
Laboratory of Systems and Synthetic Biology, Microbial Systems and Synthetic Biology, Dreijenplein 10, Wageningen 6703 HB, Netherlands.
Leo H
LH
de Graaff
Aspergillus; Gaeumannomyces graminis; Lipoxygenase; Protease; Proteomics; Thioredoxin reductase