Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli

Liang, Mingzhi; Frank, Stefanie; Lünsdorf, Heinrich; Warren, Martin J.; Prentice, Michael B.

Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli

dc.contributor.author	Liang, Mingzhi
dc.contributor.author	Frank, Stefanie
dc.contributor.author	Lünsdorf, Heinrich
dc.contributor.author	Warren, Martin J.
dc.contributor.author	Prentice, Michael B.
dc.contributor.funder	Health Research Board	en
dc.contributor.funder	Science Foundation Ireland	en
dc.contributor.funder	Biotechnology and Biological Sciences Research Council	en
dc.date.accessioned	2017-07-04T09:13:34Z
dc.date.available	2017-07-04T09:13:34Z
dc.date.issued	2017-02-10
dc.date.updated	2017-07-03T16:19:16Z
dc.description.abstract	Processes for the biological removal of phosphate from wastewater rely on temporary manipulation of bacterial polyphosphate levels by phased environmental stimuli. In E. coli polyphosphate levels are controlled via the polyphosphate-synthesizing enzyme polyphosphate kinase (PPK1) and exopolyphosphatases (PPX and GPPA), and are temporarily enhanced by PPK1 overexpression and reduced by PPX overexpression. We hypothesised that partitioning PPK1 from cytoplasmic exopolyphosphatases would increase and stabilise E. coli polyphosphate levels. Partitioning was achieved by co-expression of E. coli PPK1 fused with a microcompartment-targeting sequence and an artificial operon of Citrobacter freundii bacterial microcompartment genes. Encapsulation of targeted PPK1 resulted in persistent phosphate uptake and stably increased cellular polyphosphate levels throughout cell growth and into the stationary phase, while PPK1 overexpression alone produced temporary polyphosphate increase and phosphate uptake. Targeted PPK1 increased polyphosphate in microcompartments 8-fold compared with non-targeted PPK1. Co-expression of PPX polyphosphatase with targeted PPK1 had little effect on elevated cellular polyphosphate levels because microcompartments retained polyphosphate. Co-expression of PPX with non-targeted PPK1 reduced cellular polyphosphate levels. Thus, subcellular compartmentalisation of a polymerising enzyme sequesters metabolic products from competing catabolism by preventing catabolic enzyme access. Specific application of this process to polyphosphate is of potential application for biological phosphate removal.	en
dc.description.sponsorship	Health Research Board (award HRA_POR/2011/111); Science Foundation Ireland (SFI Grant Numbers 11/TIDA/B2001 and SFI/12/RC/2273); British Biotechnology and Biological Sciences Research Council (BB/M002969 and BB/H013180)	en
dc.description.status	Peer reviewed	en
dc.description.version	Accepted Version	en
dc.format.mimetype	application/pdf	en
dc.identifier.articleid	1600415
dc.identifier.citation	Liang, M., Frank, S., Lünsdorf, H., Warren, M. J. and Prentice, M. B. (2017) 'Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli', Biotechnology Journal, 12(3), 1600415	en
dc.identifier.doi	10.1002/biot.201600415
dc.identifier.endpage	1600415	en
dc.identifier.issn	1860-6768
dc.identifier.issued	3	en
dc.identifier.journaltitle	Journal of Biotechnology	en
dc.identifier.startpage	1600415	en
dc.identifier.uri	https://hdl.handle.net/10468/4211
dc.identifier.volume	12	en
dc.language.iso	en	en
dc.publisher	Wiley-VCH Verlag	en
dc.rights	This is the accepted version of the following article: Liang et al (2017), Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli. Biotechnol. J., 12: 1600415 which has been published in final form at http://dx.doi.org/10.1002/biot.201600415 .This article may be used for non-commercial purposes in accordance with the Wiley Self-Archiving Policy [olabout.wiley.com/WileyCDA/Section/id-820227.html]	en
dc.subject	Bacteria	en
dc.subject	Biopolymers	en
dc.subject	Metabolic engineering	en
dc.subject	Microreactors	en
dc.subject	Synthetic biology	en
dc.subject	Escherichia coli	en
dc.subject	Inorganic polyphosphate	en
dc.subject	Waste water	en
dc.subject	Salmonella enterica	en
dc.subject	Protein	en
dc.subject	Phosphate	en
dc.subject	Gene	en
dc.subject	Metabolism	en
dc.subject	Organelles	en
dc.subject	Sequences	en
dc.title	Bacterial microcompartment-directed polyphosphate kinase promotes stable polyphosphate accumulation in E. coli	en
dc.type	Article (peer-reviewed)	en