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Proteomic enzyme analysis of the marine fungus Paradendryphiella salina reveals alginate lyase as a minimal adaptation strategy for brown algae degradation

机译:对海洋真菌Paradendryphiella salina的蛋白质组酶分析表明藻酸盐裂解酶是褐藻降解的最小适应策略

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摘要

We set out to investigate the genetic adaptations of the marine fungus Paradendryphiella salina CBS112865 for degradation of brown macroalgae. We performed whole genome and transcriptome sequencing and shotgun proteomic analysis of the secretome of P. salina grown on three species of brown algae and under carbon limitation. Genome comparison with closely related terrestrial fungi revealed that P. salina had a similar but reduced CAZyme profile relative to the terrestrial fungi except for the presence of three putative alginate lyases from Polysaccharide Lyase (PL) family 7 and a putative PL8 with similarity to ascomycete chondroitin AC lyases. Phylogenetic and homology analyses place the PL7 sequences amongst mannuronic acid specific PL7 proteins from marine bacteria. Recombinant expression, purification and characterization of one of the PL7 genes confirmed the specificity. Proteomic analysis of the P. salina secretome when growing on brown algae, revealed the PL7 and PL8 enzymes abundantly secreted together with enzymes necessary for degradation of laminarin, cellulose, lipids and peptides. Our findings indicate that the basic CAZyme repertoire of saprobic and plant pathogenic ascomycetes, with the addition of PL7 alginate lyases, provide P. salina with sufficient enzymatic capabilities to degrade several types of brown algae polysaccharides.
机译:我们着手研究海洋真菌对虾褐藻CBS112865对棕色大型藻类降解的遗传适应性。我们进行了全基因组和转录组测序,并对在三种碳藻限制下生长在三种褐藻上的盐沼假单胞菌的分泌组进行了鸟枪蛋白质组分析。基因组与密切相关的陆生真菌的比较显示,与陆地真菌相比,盐沼疟原虫具有相似但降低的CAZyme谱,除了存在来自多糖裂解酶(PL)家族7的三种推定藻酸盐裂解酶和与子囊软骨素相似的推定PL8外AC裂解酶。系统发生和同源性分析将PL7序列置于来自海洋细菌的甘露糖醛酸特异性PL7蛋白之中。 PL7基因之一的重组表达,纯化和鉴定证实了特异性。对在褐色藻类上生长的盐沼假单胞菌分泌蛋白组进行蛋白质组学分析,发现PL7和PL8酶与层粘连蛋白,纤维素,脂质和肽降解所必需的酶一起大量分泌。我们的发现表明,腐殖酸和植物病原性子囊藻的基本CAZyme库,加上PL7海藻酸盐裂解酶,为盐沼假单胞菌提供了足以降解几种褐藻多糖的酶促能力。

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