Characterization of heterologous hemoglobin and flavohemoglobin promoter regulation in Escherichia coli.
Bacterial hemoglobins and flavohemoglobins have been used to improve cell growth and productivity in biotechnological applications. The expression of globin genes can be induced by reducing the oxygen supply or applying external stressors, which provide a simple and inexpensive mechanism for induction of heterologous protein production. It is in the interest of the biotechnological industry to seek new promoters, which are non-patented, cheap and simple to induce. Therefore, new globin gene promoters have been isolated from Campylobacter jejuni, Bacillus subtilis, Deinococcus radiodurans, Streptomyces coelicolor, and Salmonella typhi. The goal was to obtain insights about the regulation mechanisms of these promoters in Escherichia coli using in silico and experimental methods. The recognition of these promoters by the E. coli transcriptional machinery was first analyzed by computational methods. Computer analysis revealed that all the promoters, except the promoter of S. coelicolor, should be functional in E. coli and most of them also contain putative binding sites for ArcA, CRP, and FNR global regulators. Furthermore, the expression profiles of the promoters fused to the chloramphenicol acetyl transferase gene were analyzed under various conditions using E. coli mutants devoid of regulatory molecules. In vivo regulation studies of globin promoters mainly verified the in silico predictions.
Koskenkorva
Taija
T
Institute of Biotechnology, ETH Zürich, CH-8093 Zürich, Switzerland.
Frey
Alexander D
AD
Kallio
Pauli T
PT
D001419:Bacteria / Q000235:genetics; D002241:Carbohydrates / Q000494:pharmacology; D019295:Computational Biology; D004093:Dihydropteridine Reductase / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering; D006420:Hemeproteins / Q000096:biosynthesis* / Q000235:genetics; D006454:Hemoglobins / Q000096:biosynthesis* / Q000235:genetics; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics*
D002241:Carbohydrates; D029968:Escherichia coli Proteins; C060704:FNR protein, E coli; D006420:Hemeproteins; D006454:Hemoglobins; D007506:Iron-Sulfur Proteins; D004093:Dihydropteridine Reductase; C076538:hmp protein, E coli; D009247:NADH, NADPH Oxidoreductases