Systems Biology of Recombinant Protein Production by Fungi

摘要: Systems biology has emerged as a highly potent tool for studying biological processes over the last decades. However, its application to complex metabolic such protein secretion is still at infant stage. Saccharomyces cerevisiae and Aspergillus oryzae are two important fungal cell factories which occupy significant proportions of recombinant productions, whereas various bottlenecks undiscovered mechanisms limit their full potential robust hosts. In this thesis, systems approaches were applied explore these organisms in respect production. By utilizing engineering yeast endogenous heme synthesis, we demonstrated possibility efficient production proteins (e.g. multimer with prosthetic group) by yeast. Applying inverse engineering, identified many genomic variants that may contribute improve Specifically, examined effect single point mutation on VTA1 encoding regulatory MVB pathway endocytosis. Our result suggests VTA1S196I might help accelerate nutrient uptake via endocytosis, subsequently enhanced synthesis secretion. Oxygen an element associated normal cellular metabolism well We studied how Rox1p, heme-dependent transcription repressor hypoxia-induced genes, affect yeast, under aerobic conditions. knocking out ROX1, observed 100% increase α-amylase Through genome wide transcriptome analysis several Rox1p targets based suggested roles improving productions. Lastly, applying comparative genomics study, enriched list core components involved secretory machinery A. oryzae. To verify list, high producing strains constructed. The transcriptional responses using microarray, through strategies including overexpressing up-regulated wall EglD Cwp1 genes extracellular competing pathway, proposed.