Synthetic control devices for gene regulation in Penicillium chrysogenum
Journal article, 2019

BACKGROUND: Orthogonal, synthetic control devices were developed for Penicillium chrysogenum, a model filamentous fungus and industrially relevant cell factory. In the synthetic transcription factor, the QF DNA-binding domain of the transcription factor of the quinic acid gene cluster of Neurospora crassa is fused to the VP16 activation domain. This synthetic transcription factor controls the expression of genes under a synthetic promoter containing quinic acid upstream activating sequence (QUAS) elements, where it binds. A gene cluster may demand an expression tuned individually for each gene, which is a great advantage provided by this system. RESULTS: The control devices were characterized with respect to three of their main components: expression of the synthetic transcription factors, upstream activating sequences, and the affinity of the DNA binding domain of the transcription factor to the upstream activating domain. This resulted in synthetic expression devices, with an expression ranging from hardly detectable to a level similar to that of highest expressed native genes. The versatility of the control device was demonstrated by fluorescent reporters and its application was confirmed by synthetically controlling the production of penicillin. CONCLUSIONS: The characterization of the control devices in microbioreactors, proved to give excellent indications for how the devices function in production strains and conditions. We anticipate that these well-characterized and robustly performing control devices can be widely applied for the production of secondary metabolites and other compounds in filamentous fungi.

Synthetic expression system

Gene regulation

Secondary metabolite production

Hybrid transcription factor

Synthetic gene cluster

Penicillium chrysogenum

Author

László Mózsik

University of Groningen

Zsófia Büttel

University of Groningen

R. A. L. Bovenberg

DSM Biotechnology Centre

University of Groningen

Arnold J.M. Driessen

University of Groningen

Yvonne Nygård

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

University of Groningen

DSM Biotechnology Centre

Microbial Cell Factories

1475-2859 (ISSN)

Vol. 18 1 203-

Subject Categories

Biochemistry and Molecular Biology

Microbiology

Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

DOI

10.1186/s12934-019-1253-3

PubMed

31739777

More information

Latest update

11/26/2019