Secretome analysis of the basidiomycete Pleurotus sapidus. Secretome of Pleurotus sapidus

A kinetic analysis of the secretome of P. sapidus was performed over the culture period of 12 days. Extracellular enzyme activities were determined in surface and submerged cultures, and more than 200 protein spots were separated by means of high resolution two-dimensional gel electrophoresis. Peptides of 144 proteins were tentatively identified by MALDI-MS/MS analyses. Though P. sapidus is a white-rot fungus, ~40% of the identified proteins were enzymes acting on carbohydrates. 28% of the proteins represented members of the glycoside hydrolase family, 7% carbohydrate esterases, and 5% polysaccharide lyases. Additionally, peptidases (8.3%) and oxidoreductases (8.3%) were identified. Complete cDNA sequences encoding various enzymes of the secretome have been cloned.

本研究在为期12天的培养周期内，对P. sapidus的分泌组（secretome）开展了动力学分析。研究测定了其表面培养与深层液体培养体系中的胞外酶活性，通过高分辨率二维凝胶电泳分离得到200余个蛋白质斑点。经基质辅助激光解吸电离串联质谱（MALDI-MS/MS）分析，对144种蛋白质的肽段进行了初步鉴定。尽管P. sapidus属于白腐真菌，但所鉴定的蛋白质中约40%为作用于碳水化合物的酶类：其中28%为糖苷水解酶家族成员，7%为碳水化合物酯酶，5%为多糖裂解酶；此外还鉴定出占比8.3%的肽酶与8.3%的氧化还原酶。本研究已克隆得到编码该分泌组中多种酶的完整互补DNA（cDNA）序列。