Contributes for the development of Ashbya gossypii as a cell factory

Abstract

[Excerpt] Ashbya gossypii is a filamentous Saccharomycete long known by the scientific and industrial communities, first as a cotton pathogen and subsequently as a riboflavin overproducer. Its industrial relevance combined with its high genetic similarity with Saccharomyces cerevisiae promoted the development of a significant molecular and in silico toolbox for its genetic engineering. This, together with the increasing knowledge of its genome, transcriptome and metabolism has helped designing effective metabolic engineering strategies for optimizing riboflavin production, and also for developing new A. gossypii strains for novel biotechnological applications [1]. Here, we will address our main contributes for the development of A. gossypii as a cell factory organism, by presenting an overview of the most representative outputs from our research [referenced in 1, 2]. Envisioning its exploration as a recombinant protein producer, our main efforts focused on the characterization of the A. gossypii protein secretory pathway at the genomic, transcriptomic and proteomic levels [2]. Based on experimental observations and on data from omic analyses, a hydrolytic enzyme, invertase, was deduced to be natively secreted by A. gossypii and molecularly characterized. The N-glycosylation pattern of the proteins natively secreted by A. gossypii was also characterized, as well as the recombinant production by this fungus of secreted proteins from different fungal origins. Among these, the β-galactosidase from Aspergillus niger was expressed in A. gossypii under the regulation of several native and heterologous promoters, presenting the highest extracellular production levels. [...]