A closer look at Aspergillus: Online monitoring via scattered light enables reproducible phenotyping

Development of microtiter plate based microbioreactor cultivation for Aspergillus giganteus with quasi-continuous online measurements. Different parameters such as well geometry, shaking frequency and morphology controlling agents were investigated in order to optimize the microtiter plate cultivation and scattered light signal towards reproducibility and homogeneity. An optimized medium was developed and scalability into stirred tank bioreactor cultivation was analyzed. As a transferability indicator the supernatant of both cultivation systems was analyzed for secreted protein patterns with a focus on an antifungal protein (AFP) and alpha-sarcin. These proteins were identified via LC-MS/MS.

本研究针对巨大曲霉（Aspergillus giganteus）构建了基于微孔板（microtiter plate）的微型生物反应器培养体系，并实现准连续在线监测。为优化该微孔板培养体系并提升散射光信号的重现性与均一性，研究考察了孔板几何构型、振荡频率及形态调控剂等多项参数。本研究研发得到优化后的培养基，并分析了该体系向搅拌罐式生物反应器培养的可放大性。以可转移性为评价指标，对两种培养体系的上清液开展分泌蛋白谱分析，重点关注抗真菌蛋白（AFP）与α-菌素（alpha-sarcin）。上述蛋白均通过液相色谱-串联质谱（LC-MS/MS）完成鉴定。