Dionisio Giuseppe, Kryger Per, Steenberg Tove
Department of Molecular Biology, Aarhus University, Flakkebjerg, Forsøgsvej 1, 4200 Slagelse, Denmark.
Department of Agroecology-Entomology and Plant Pathology, Aarhus University, Flakkebjerg, Forsøgsvej 1, 4200 Slagelse, Denmark.
Insects. 2016 Oct 14;7(4):54. doi: 10.3390/insects7040054.
is an entomopathogenic fungus that grows both in vivo and in vitro. In vivo it can colonize live insect hosts, and tissue digestion occurs by secreted hydrolytic exoenzymes. It can also colonize dead insect tissue provided this is free from competing microorganisms. Depending on whether the host is alive or dead the expression (quality/quantity) of the exoenzymes may vary. We have grown several isolates of in shaking flasks for 120 h at 25 °C in order to evaluate the maximal exoenzyme production using two diet regimes. As sole carbon, nitrogen, and phosphate sources we used 1% shrimp chitin and either 0.5% / of dead intact American cockroach () or their isolated cuticles. This is the first report of a differential proteomics of exoenzymes performed by label-free nano-LC MS/MS. Total proteolytic enzyme activity was mainly due to Pr1A or Pr1B depending on the isolate and the diet regime. The most differentially secreted enzymes were: the cuticle-degrading subtilisin Pr1A, GH13 alpha-glycosidase, glucan endo-1,3-beta-glucosidase, subtilisin-like proteinase Spm1, lipase 1, beta-1,3 exoglucanase, and endo-1,3-beta-glucosidase. Among the isolates analyzed, Bb 678 and Bb BG were the most active in Pr1A secretion.
是一种既能在体内生长也能在体外生长的昆虫病原真菌。在体内,它可以定殖于活的昆虫宿主,通过分泌的水解性胞外酶进行组织消化。它也可以定殖于死亡的昆虫组织,前提是该组织没有竞争性微生物。根据宿主是活的还是死的,胞外酶的表达(质量/数量)可能会有所不同。我们在25℃下于摇瓶中培养了几种该真菌的分离株120小时,以便使用两种饮食方案评估最大胞外酶产量。作为唯一的碳、氮和磷源,我们使用了1%的虾壳聚糖以及0.5%完整的死美洲大蠊()或其分离的表皮。这是通过无标记纳米液相色谱串联质谱法对该真菌胞外酶进行差异蛋白质组学研究的首次报告。总蛋白酶活性主要归因于Pr1A或Pr1B,这取决于分离株和饮食方案。分泌差异最大的酶有:角质层降解枯草杆菌蛋白酶Pr1A、GH13α-糖苷酶、葡聚糖内切-1,3-β-葡糖苷酶、类枯草杆菌蛋白酶Spm1、脂肪酶1、β-1,3外切葡聚糖酶和内切-1,3-β-葡糖苷酶。在所分析的分离株中,Bb 678和Bb BG在Pr1A分泌方面最为活跃。