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|Title:||In vitro conversion of penicillin G and ampicillin by recombinant Streptomyces clavuligerus NRRL 3585 deacetoxycephalosporin C synthase||Authors:||Sim, J.
Deacetoxycephalosproin C synthase
|Issue Date:||2001||Citation:||Sim, J., Sim, T.-S. (2001). In vitro conversion of penicillin G and ampicillin by recombinant Streptomyces clavuligerus NRRL 3585 deacetoxycephalosporin C synthase. Enzyme and Microbial Technology 29 (4-5) : 240-245. ScholarBank@NUS Repository.||Abstract:||The in vitro conversion of penicillin N analogues to cephalosporins was first demonstrated in resting cells and cell-free extracts of Streptomyces clavuligerus NP1. This study has optimized the conversion rate of a highly expressed soluble source of recombinant S. clavuligerus NRRL 3585 deacetoxycepalosporin C synthase (scDAOCS) through detailed characterization of its cofactor requirements. Inclusion of dithiothreitol (DTT) and ascorbate at various concentrations from 0.2-16 mM and 0.2-10 mM was found to be inhibitory to scDAOCS activity under specified reaction conditions examined. Simultaneous omission of these cofactors in a new modified reaction condition, comprising of 50 mM Tris HCl (pH7.4), 0.8 mM ATP, 1.28 mM α-ketoglutarate and 1.8 mM FeSO4, enhanced the conversion rate of two penicillin N analogues, penicillin G and ampicillin, through 20- and 35-fold respectively. © 2001 Elsevier Science Inc. All rights reserved.||Source Title:||Enzyme and Microbial Technology||URI:||http://scholarbank.nus.edu.sg/handle/10635/31108||ISSN:||01410229|
|Appears in Collections:||Staff Publications|
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