title: An improved bioprocess for synthesis of acetohydroxamic acid using DTT (dithiothreitol) treated resting cells of Bacillus sp. APB-6.
abstract: Acyltransferase activity of amidase from Bacillus sp. APB-6 was enhanced (24 U) by multiple feedings of N-methylacetamide (70 mM) into the production medium. Hyperinduced whole resting cells of Bacillus sp. APB-6 corresponding to 4 g/L (dry cell weight), when treated with 10mM DTT (dithiothreitol) resulted in 93% molar conversion of acetamide (300 mM) to acetohydroxamic acid in presence of hydroxylamine-HCl (800 mM) after 30 min at 45°C in a 1L reaction mixture. After lyophilization, a 62 g powder containing 34% (wtwt(-1)) acetohydroxamic acid was recovered. This is the first report where DTT has been used to enhance acyltransfer reaction and such high molar conversion (%) of amide to hydroxamates was recorded at 1L scale.
authors.affiliation: Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla 171005, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Pandey
authors.forename: Rajendra
authors.initials: R
authors.lastname: Singh
authors.forename: Duni
authors.initials: D
authors.lastname: Chand
mesh_terms: D000081:Acetamides / Q000378:metabolism; D000217:Acyltransferases / Q000096:biosynthesis; D000581:Amidohydrolases / Q000378:metabolism; D001407:Bacillus / Q000166:cytology* / Q000187:drug effects* / Q000254:growth & development; D001709:Biotechnology / Q000379:methods*; D003470:Culture Media / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006877:Hydroxamic Acids / Q000138:chemical synthesis*; D019811:Hydroxylamine / Q000378:metabolism; D007477:Ions; D008670:Metals / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D059027:Recycling; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature; D013997:Time Factors
chemical_list: D000081:Acetamides; D003470:Culture Media; D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D007477:Ions; D008670:Metals; D009570:Nitriles; D019811:Hydroxylamine; C006358:acetohydroxamic acid; D000217:Acyltransferases; D000581:Amidohydrolases; C019665:amidase; D004229:Dithiothreitol