Purification and initial characterization of proline 4-hydroxylase from Streptomyces griseoviridus P8648: A 2-oxoacid, ferrous-dependent dioxygenase involved in etamycin biosynthesis

Lawrence, Christopher C., Sobey, Wendy J., Field, Robert A. ORCID: https://orcid.org/0000-0001-8574-0275, Baldwin, Jack E. and Schofield, Christopher J. (1996) Purification and initial characterization of proline 4-hydroxylase from Streptomyces griseoviridus P8648: A 2-oxoacid, ferrous-dependent dioxygenase involved in etamycin biosynthesis. Biochemical Journal, 313 (1). pp. 185-191. ISSN 0264-6021

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Abstract

Proline 4-hydroxylase is a 2-oxoacid, ferrous-ion-dependent dioxygenase involved in the biosynthesis of the secondary metabolite etamycin. The purification, in low yield, of proline 4-hydroxylase from Streptomyces griseoviridis P8648 to near apparent homogeneity and its initial characterization are reported. In most respects proline 4-hydroxylase is a typical member of the 2-oxoacid-dependent dioxygenase family. It is monomeric (M(r) approx. 38000) (by gel filtration on Superdex-G75) and has typically strict requirements for ferrous ion and 2-oxoglutarate. The enzyme was inhibited by aromatic analogues of 2-oxoglutarate. L-Proline-uncoupled turnover of 2-oxoglutarate to succinate and CO2 was observed. The addition of L-ascorbate did not stimulate L-proline-coupled turnover of 2-oxoglutarate, but did stimulate L-proline-uncoupled turnover. L-Ascorbate caused a time-dependent inhibition of L-proline hydroxylation. The enzyme was completely inactivated by preincubation with diethyl pyrocarbonate under histidine-modifying conditions. This inactivation could be partially prevented by the inclusion of L-proline and 2-oxoglutarate in the preincubation mixture, suggesting the presence of histidine residue(s) at the active site.

Item Type:	Article
Uncontrolled Keywords:	biochemistry,molecular biology,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1303
Faculty \ School:	Faculty of Science > School of Chemistry, Pharmacy and Pharmacology
Related URLs:	http://www.scopus.com/inward/record.url?...
Depositing User:	LivePure Connector
Date Deposited:	11 Sep 2024 14:30
Last Modified:	25 Sep 2024 18:09
URI:	https://ueaeprints.uea.ac.uk/id/eprint/96714
DOI:	10.1042/bj3130185

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