Monday, November 9, 2009
P14

Improving Aspergillus niger as a production host through manipulation of pH responding transcription factors

Lars Poulsen1, Kenneth S. Bruno2, Jette Thykaer1, Scott E. Baker2, and Anna Eliasson Lantz1. (1) Department of Systems Biology, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark, (2) Fungal Biotechnology Team, Pacific Northwest National Laboratory, Richland, WA 99354

Altering fluxes for overcoming metabolic bottlenecks have traditionally been approached by genetic engineering of a single or few metabolic genes. This strategy struggles to overcome the subjacent regulation and is thus frequently of limited success. Transcription factors have the potential of controlling several fluxes, hence manipulating expression of these proteins can provide an alternative tool for overcoming metabolic bottlenecks. In the present study the effect of modulation of transcription factors in Aspergillus niger, which is an industrially important micro-organism used in various processes including organic acid and enzyme production, was investigated. The strategy described in this work focuses on regulation connected to pH. It was chosen as an important process parameter, due to its significant influences on both organic acid and enzyme production. A previous transcription analysis identified several putative transcription factors with pH responding behavior. A number of these genes were selected as targets for gene knockout. The resulting mutants were first exposed to screening experiments including morphological studies and investigation of acid profile and protease activity. Among others an interesting finding was that one mutant had an oxalic acid overproducing phenotype (OOP). In the screening experiments the OOP mutant showed a 30 % increase in oxalic acid titer. The OOP mutant was further characterized in 2L scale bioreactors, and a 90% increase of the overall yield coefficient of oxalic acid on glucose was seen. Further data on the OOP mutant will be presented and results from the screening experiments and other mutants of industrial interest discussed.