Mutational analysis and characterization of nocardicin C-9' epimerase

The Journal of Biological Chemistry
Wendy L Kelly, Craig A Townsend

Abstract

The biosynthetic gene cluster for the nocardicin A producer Nocardia uniformis subsp. tsuyamanensis ATCC 21806 was recently identified. Nocardicin A is the most potent of a series of monocyclic beta-lactam antibiotics produced by this organism. Its activity has been attributed to a syn-configured oxime moiety and a d-homoseryl side chain attached through an unusual ether linkage to the core nocardicin framework. Notably present in the nocardicin biosynthetic gene cluster is nocJ, encoding a protein with sequence similarity to the pyridoxal 5'-phosphate (PLP)-dependent 1-aminocyclopropane-1-carboxylic acid deaminases. Insertional mutagenesis of nocJ abolished nocardicin A production, while the l-homoseryl isomer, isonocardicin A, was still observed. Expression of the disrupted nocJ gene in trans was sufficient to restore production of nocardicin A in the disruption mutant. Heterologous expression, purification, and in vitro characterization of NocJ by UV spectroscopy, cofactor reduction, chiral HPLC analysis of the products and their exchange behavior in deuterium oxide led to confirmation of its role as the PLP-dependent nocardicin C-9' epimerase responsible for interconversion of the nocardicin homoseryl side chain in both noc...Continue Reading

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Citations

Mar 1, 2007·Indian Journal of Microbiology·S Malhotra, R Lal
Aug 23, 2005·Chemical Communications : Chem Comm·Nadia J KershawChristopher J Schofield
Feb 18, 2014·Nature Chemical Biology·Nicole M Gaudelli, Craig A Townsend
Dec 19, 2008·Chembiochem : a European Journal of Chemical Biology·Refaat B HamedChristopher J Schofield
Nov 9, 2012·Natural Product Reports·Refaat B HamedChristopher J Schofield
Aug 18, 2017·Medicinal Research Reviews·Lena DecuyperStanislav Gobec
Jan 20, 2019·Journal of Industrial Microbiology & Biotechnology·Dipesh DhakalJae Kyung Sohng

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