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Heterologous expression of Gaeumannomyces graminis lipoxygenase in Aspergillus nidulans

机译:禾本科牛油杆菌脂加氧酶在构巢曲霉中的异源表达

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摘要

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.
机译:曲霉包含编码Ppo酶的ppo基因,该酶可从多不饱和脂肪酸产生脂蛋白。这些脂蛋白在孢子形成中充当信号分子,并影响曲霉菌的无性与有性比。像构巢曲霉和黑曲霉这样的真菌仅含有ppo基因,而人类致病性黄曲霉和烟曲霉则含有ppo基因和脂氧合酶。脂氧合酶催化脂氧合蛋白的合成,并被认为与群体感应能力和入侵植物组织有关。在这项研究中,我们使用构巢曲霉WG505作为表达宿主,异源表达Gaeumannomyces graminis脂加氧酶。重组LOX的存在会诱导构巢曲霉转化子发生表型变化。而且,对构巢曲霉LOX产生菌株的蛋白质组学分析表明,异源蛋白在其分泌途径中的糖基化之前被降解。我们观察到,LOX的存在会诱导氨基肽酶Y的特定产生,这种酶可能会在细胞间降解格里米斯芽孢杆菌脂氧合酶。同样,蛋白硫氧还蛋白还原酶的存在表明,在构巢曲霉中,格里米纳斯氏菌脂氧化酶被积极抑制。

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