Highly purified E. coli DNA Gyrase enzyme (activity-validated) is offered as a holoenzyme (contains both A,B subunits). This purified type II topoisomerase is designed for reliable in vitro DNA supercoiling assays and antibacterial inhibitor screening. E. coli DNA Gyrase efficiently converts relaxed plasmid DNA into supercoiled DNA in an ATP-dependent reaction, making it ideal for mechanistic studies, assay development, and compound profiling.
Common applications include DNA topology research, gyrase functional validation, and screening of compounds to distinguish gyrase poisons vs. catalytic inhibitors. Our enzyme is a dependable choice for academic, biotech, and pharmaceutical R&D teams requiring reproducible gyrase performance and clean, interpretable assay outcomes.
Use of this enzyme product enables
- Validated enzymology (clean gyrase + defined substrates)
- MOA deconvolution (poisons vs catalytic inhibitors)
- Workflow acceleration (rapid gel-based readouts; screening-friendly format)
- Real discovery outcomes
*new toxin biology impacting gyrase
*new inhibitor families hitting gyrase
*assay tech development for throughput
E. coli DNA Gyrase Quality Control Tests
Nuclease contamination: assayed by testing for linear KDNA and linear plasmid DNA formation. Incubation of 1 µg of catenated KDNA or supercoiled pUC19 DNA for 4 hrs. at 37°C (under gyrase assay conditions and with or without ATP).
Assay Conditions
Supercoiling assays are carried out using relaxed DNA under conditions specified in the protocol sheets provided with the product. We also perform assays using kDNA as substrate. For details on this decatenation/supercoiling assay please refer to our technical literature. One unit of gyrase activity will supercoil 0.2 ug of plasmid in 30 min at 37°C.
Included Materials
A 5X assay buffer and dilution buffer is included with the enzyme. These are also sold separately as 5X Gyrase Assay Buffer (TG4030) Gyrase Dilution Buffer (please inquire).
Purification of Gyrase A and B Subunits
DNA gyrase is shipped on dry ice and should be stored at -70°C. We also recommend that the enzyme be aliquoted after the first thaw (repeated rounds of freeze/thaw may cause loss of activity); the enzyme activity is stable for 1-3 days on ice.
E. coli DNA Gyrase Data Sheet
Material Safety Data Sheet
Reviews and Citations
- María-José Ferrándiz, Pablo Hernández, Adela G de la Campa: Distribution of DNA gyrase cleavage sites across the Streptococcus pneumoniae genome: relation to transcription and methylation at GATC sites. Nucleic Acids Research, 2025 Nov 20;53(21):gkaf1183. https://doi.org/10.1093/nar/gkaf1183
- Anthony Maxwell, Nicolas P Burton, Natasha O’Hagan: High-throughput assays for DNA gyrase and other topoisomerases. Nucleic Acids Research, 2006;34(15):e104. https://doi.org/10.1093/nar/gkl504
- D. Gadelle , C. Bocs , M. Graille , P. Forterre: Inhibition of archaeal growth and DNA topoisomerase VI activities by the Hsp90 inhibitor radicicol. Nucleic Acids Research, Volume 33, Issue 7, 1 April 2005, Pages 2310–2317, https://doi.org/10.1093/nar/gki526
- Phillips JW, Goetz MA, Smith SK, Zink DL, Polishook J, Onishi R, Salowe S, Wiltsie J, Allocco J, Sigmund J, Dorso K, Lee S, Skwish S, de la Cruz M, Martin J, Vicente F, Genilloud O, Lu J, Painter RE, Young K, Overbye K, Donald RGK, Singh SB: Discovery of Kibdelomycin, A Potent New Class of Bacterial Type II Topoisomerase Inhibitor by Chemical-Genetic Profiling in Staphylococcus aureus. Cell Chemistry and Biology 2011, 18: 955-965. doi: 10.1016/j.chembiol.2011.06.011.
