Hasan, Muhammad, Fayter, Alice E. R. and Gibson, Matthew I. (2018) Ice recrystallization inhibiting polymers enable glycerol-free cryopreservation of microorganisms. Biomacromolecules, 19 (8). pp. 3371-3376. doi:10.1021/acs.biomac.8b00660 ISSN 1525-7797.

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Abstract

All modern molecular biology and microbiology is underpinned by not only the tools to handle and manipulate microorganisms but also those to store, bank, and transport them. Glycerol is the current gold-standard cryoprotectant, but it is intrinsically toxic to most microorganisms: only a fraction of cells survive freezing and the presence of glycerol can impact downstream applications and assays. Extremophile organisms survive repeated freeze/thaw cycles by producing antifreeze proteins which are potent ice recrystallization inhibitors. Here we introduce a new concept for the storage/transport of microorganisms by using ice recrystallization inhibiting poly(vinyl alcohol) in tandem with poly(ethylene glycol). This cryopreserving formulation is shown to result in a 4-fold increase in E. coli yield post-thaw, compared to glycerol, utilizing lower concentrations, and successful cryopreservation shown as low as 1.1 wt % of additive. The mechanism of protection is demonstrated to be linked not only to inhibiting ice recrystallization (by comparison to a recombinant antifreeze protein) but also to the significantly lower toxicity of the polymers compared to glycerol. Optimized formulations are presented and shown to be broadly applicable to the cryopreservation of a panel of Gram-negative, Gram-positive, and mycobacteria strains. This represents a step-change in how microorganisms will be stored by the design of new macromolecular ice growth inhibitors; it should enable a transition from traditional solvent-based to macromolecular microbiology storage methods.

Item Type:	Journal Article
Subjects:	Q Science > QH Natural history > QH301 Biology Q Science > QR Microbiology
Divisions:	Faculty of Science, Engineering and Medicine > Science > Chemistry
Library of Congress Subject Headings (LCSH):	Cryobiology, Microorganisms, Polymers
Journal or Publication Title:	Biomacromolecules
Publisher:	American Chemical Society
ISSN:	1525-7797
Official Date:	13 August 2018
Dates:	Date Event 13 August 2018 Published 22 June 2018 Available 22 June 2018 Accepted
Volume:	19
Number:	8
Page Range:	pp. 3371-3376
DOI:	10.1021/acs.biomac.8b00660
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Open Access (Creative Commons)
Date of first compliant deposit:	31 July 2018
Date of first compliant Open Access:	31 July 2018
RIOXX Funder/Project Grant:	Project/Grant ID RIOXX Funder Name Funder ID 638661 European Research Council http://viaf.org/viaf/130022607 UNSPECIFIED [RS] Royal Society http://dx.doi.org/10.13039/501100000288 BB/M01116X/1 [BBSRC] Biotechnology and Biological Sciences Research Council http://dx.doi.org/10.13039/501100000268
Related URLs:	Related dataset

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