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Aminoglycoside antibiotics become inactivated by aminoglycoside-6\u2032-N-acetyltransferase-Ib [AAC(6\u2032)-Ib] of gram-negative bacteria which transfers an acetyl group from acetyl-CoA to the antibiotic. The aim of the study was to disrupt the enzymatic activity of AAC(6\u2032)-Ib by adjuvants and restore aminoglycoside activity as a result. The binding affinities of several vitamins and chemical compounds with AAC(6\u2032)-Ib of<jats:italic>Escherichia coli<\/jats:italic>,<jats:italic>Klebsiella pneumoniae<\/jats:italic>, and<jats:italic>Shigella sonnei<\/jats:italic>were determined by molecular docking method to screen potential adjuvants. Adjuvants having higher binding affinity with target enzymes were further analyzed in-vitro to assess their impact on bacterial growth and bacterial modifying enzyme AAC(6\u2032)-Ib activity. Four compounds\u2014zinc pyrithione (ZnPT), vitamin D, vitamin E and vitamin K-exhibited higher binding affinity to AAC(6\u2032)-Ib than the enzyme\u2019s natural substrate acetyl-CoA. Combination of each of these adjuvants with three aminoglycoside antibiotics\u2014amikacin, gentamicin and kanamycin\u2014were found to significantly increase the antibacterial activity against the selected bacterial species as well as hampering the activity of AAC(6\u2032)-Ib. The selection process of adjuvants and the use of those in combination with aminoglycoside antibiotics promises to be a novel area in overcoming bacterial resistance.<\/jats:p>","DOI":"10.1038\/s41598-020-76355-0","type":"journal-article","created":{"date-parts":[[2020,11,9]],"date-time":"2020-11-09T11:05:02Z","timestamp":1604919902000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Retention of antibiotic activity against resistant bacteria harbouring aminoglycoside-N-acetyltransferase enzyme by adjuvants: a combination of in-silico and in-vitro study"],"prefix":"10.1038","volume":"10","author":[{"given":"Shamim","family":"Ahmed","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina Amita","family":"Sony","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md. 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