I am working on a multi-enzyme product stability study and worried protease might interfere with the detection rest of the enzymes. Is there a Bacillolysin protease inhibitor that I can use? I am testing on the UV spec.
The BRENDA database lists a substance called N-[alpha-L-rhamnopyranosyl-oxyhydroxyphosphinyl]-L-leucyl-L-tryptophan as an inhibitor of bacillolysin, with a Ki of 3.5 µM.
https://brenda-enzymes.org/enzyme.php?ecno=3.4.24.28#INHIBITOR
https://brenda-enzymes.org/literature.php?e=3.4.24.28&r=650655
I don't know anything about your multi-enzyme system, but bacillolysin is a metalloprotease so should be strongly inhibited by metal chelators like EDTA and EGTA at 1 mM. However, this would also inhibit other metallozymes in your system.
If that's a problem, there are also dozens of commercially available protease inhibitor cocktails purpose-designed to inhibit multiple protease activities (i.e. Ser/Cys and Glu/Asp) without chelating metals so the downstream solution can be purified by metal affinity chromatography - i.e. https://www.sigmaaldrich.com/CA/en/product/roche/04693159001
Alexander C. Anderson Thank you so much the EDTA Di-Sodium salt will work perfectly since there other enzymes in the product are non-metalloenzymes.
Adam B Shapiro Thanks I will look them up and if I manage to get them on time, I can actually use both approaches and do comparison studies.