Evaluation of NfsA-like nitroreductases from <italic>Neisseria meningitidis</italic> and <italic>Bartonella henselae</italic> for enzyme-prodrug therapy, targeted cellular ablation, and dinitrotoluene bioremediation.

Rich, Michelle H.; Sharrock, Abigail V.; Hall, Kelsi R.; Ackerley, David F.; MacKichan, Joanna K.

Objectives: To characterize the activities of two candidate nitroreductases, Neisseria meningitidis NfsA (NfsA_Nm) and Bartonella henselae (PnbA_Bh), with the nitro-prodrugs, CB1954 and metronidazole, and the environmental pollutants 2,4- and 2,6-dinitrotoluene.Results: NfsA_Nm and PnbA_Bh were evaluated in Escherichia coli over-expression assays and as His6-tagged proteins in vitro. With the anti-cancer prodrug CB1954, both enzymes were more effective than the canonical O2-insensitive nitroreductase E. coli NfsB (NfsB_Ec), NfsA_Nm exhibiting comparable levels of activity to the leading nitroreductase candidate E. coli NfsA (NfsA_Ec). NfsA_Nm is also the first NfsA-family nitroreductase shown to produce a substantial proportion of 4-hydroxylamine end-product. NfsA_Nm and PnbA_Bh were again more efficient than NfsB_Ec at aerobic activation of metronidazole to a cytotoxic form, with NfsA_Nm appearing a promising candidate for improving zebrafish-targeted cell ablation models. NfsA_Nm was also more active than either NfsA_Ec or NfsB_Ec with 2,4- or 2,6-dinitrotoluene substrates, whereas PnbA_Bh was relatively inefficient with either substrate.Conclusions: NfsA_Nm is a promising new nitroreductase candidate for several diverse biotechnological applications.

NEISSERIA meningitidis; NITROREDUCTASES; BARTONELLA henselae; PRODRUGS; METRONIDAZOLE

Biotechnology Letters, 2018, Vol 40, Issue 2, p359

0141-5492

Academic Journal

10.1007/s10529-017-2472-5