Streptomyces lividans displays a distinct dependence on copper to fully initiate morphological development. Evidence has accumulated to implicate the participation of an extracytoplasmic cuproenzyme in morphogenesis. In the present study, we show that GlxA fulfils all criteria to be that cuproenzyme. GlxA is membrane associated and has an active site consisting of a mononuclear copper and a cross-linked Y-C cofactor. The domain organization of the tertiary structure defines GlxA as a new structural member of the mono-copper oxidase family, with copper co-ordination geometry similar to, but spectroscopically distinct from fungal galactose oxidase (Gox). EPR spectroscopy reveals that the oxidation of cupric GlxA generates a protein radical residing on the Y-C cross-link. A variety of canonical Gox substrates (including D-galactose) were tested but none were readily turned over by GlxA. A glxA null-mutant leads to loss of glycan accumulation at hyphal tips and consequently a drastically changed morphology both on solid substrates and in liquid-grown environments, a scenario similarly observed in the absence of the neighbouring glycan synthase CslA (cellulase synthase-like protein). In addition the glxA mutant has lost the stimulation of development by copper, supporting a model whereby the enzymatic action of GlxA on the glycan is required for development and morphology. From a biotechnology perspective, the open mycelium morphology observed with the glxA mutant in submerged culture has implications for use as an enzyme production host.
Skip Nav Destination
Article navigation
August 2015
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
July 23 2015
GlxA is a new structural member of the radical copper oxidase family and is required for glycan deposition at hyphal tips and morphogenesis of Streptomyces lividans
Amanda K. Chaplin;
Amanda K. Chaplin
*School of Biological Science, University of Essex, Wivenhoe Park, Colchester, CO4 3SQ, U.K.
Search for other works by this author on:
Marloes L.C. Petrus;
Marloes L.C. Petrus
†Molecular Biotechnology, Institute of Biology, Sylvius Laboratory, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands
Search for other works by this author on:
Giulia Mangiameli;
Giulia Mangiameli
†Molecular Biotechnology, Institute of Biology, Sylvius Laboratory, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands
Search for other works by this author on:
Michael A. Hough;
Michael A. Hough
*School of Biological Science, University of Essex, Wivenhoe Park, Colchester, CO4 3SQ, U.K.
Search for other works by this author on:
Dimitri A. Svistunenko;
Dimitri A. Svistunenko
*School of Biological Science, University of Essex, Wivenhoe Park, Colchester, CO4 3SQ, U.K.
Search for other works by this author on:
Peter Nicholls;
Peter Nicholls
*School of Biological Science, University of Essex, Wivenhoe Park, Colchester, CO4 3SQ, U.K.
Search for other works by this author on:
Dennis Claessen;
Dennis Claessen
†Molecular Biotechnology, Institute of Biology, Sylvius Laboratory, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands
Search for other works by this author on:
Erik Vijgenboom;
Erik Vijgenboom
1
†Molecular Biotechnology, Institute of Biology, Sylvius Laboratory, Leiden University, PO Box 9505, 2300RA Leiden, The Netherlands
1Correspondence may be addressed to either author (email jworrall@essex.ac.uk or vijgenbo@biology.leidenuniv.nl).
Search for other works by this author on:
Jonathan A.R. Worrall
Jonathan A.R. Worrall
1
*School of Biological Science, University of Essex, Wivenhoe Park, Colchester, CO4 3SQ, U.K.
1Correspondence may be addressed to either author (email jworrall@essex.ac.uk or vijgenbo@biology.leidenuniv.nl).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
February 12 2015
Revision Received:
May 27 2015
Accepted:
June 01 2015
Accepted Manuscript online:
June 10 2015
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 2015 Authors; published by Portland Press Limited
2015
Biochem J (2015) 469 (3): 433–444.
Article history
Received:
February 12 2015
Revision Received:
May 27 2015
Accepted:
June 01 2015
Accepted Manuscript online:
June 10 2015
Citation
Amanda K. Chaplin, Marloes L.C. Petrus, Giulia Mangiameli, Michael A. Hough, Dimitri A. Svistunenko, Peter Nicholls, Dennis Claessen, Erik Vijgenboom, Jonathan A.R. Worrall; GlxA is a new structural member of the radical copper oxidase family and is required for glycan deposition at hyphal tips and morphogenesis of Streptomyces lividans. Biochem J 1 August 2015; 469 (3): 433–444. doi: https://doi.org/10.1042/BJ20150190
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.