Genetic toolbox for controlled expression of functional proteins in Geobacillus spp.
Species of genus Geobacillus are thermophilic bacteria and play an ever increasing role as hosts for biotechnological applications both in academia and industry. Here we screened a number of Geobacillus strains to determine which industrially relevant carbon sources they can utilize. One of the strains, G. thermoglucosidasius C56-YS93, was then chosen to develop a toolbox for controlled gene expression over a wide range of levels. It includes a library of semi-synthetic constitutive promoters (76-fold difference in expression levels) and an inducible promoter from the xylA gene. A library of synthetic in silico designed ribosome binding sites was also created for further tuning of translation. The PxylA was further used to successfully express native and heterologous xylanases in G. thermoglucosidasius. This toolbox enables fine-tuning of gene expression in Geobacillus species for metabolic engineering approaches in production of biochemicals and heterologous proteins.
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kgs. Lyngby, Denmark.
Ivan
I
Pogrebnyakov
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kgs. Lyngby, Denmark.
Christian Bille
CB
Jendresen
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kgs. Lyngby, Denmark.
Alex Toftgaard
http://orcid.org/0000-0001-6616-0187
AT
Nielsen
D001426:Bacterial Proteins / Q000378:metabolism; D003001:Cloning, Molecular; D003198:Computer Simulation; D003470:Culture Media; D043364:Endo-1,4-beta Xylanases / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D056506:Geobacillus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics
D001426:Bacterial Proteins; D003470:Culture Media; D043364:Endo-1,4-beta Xylanases