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An investigation into the preservation of microbial cell banks for α-amylase production during 5 l fed-batch Bacillus licheniformis fermentations

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Abstract

Fluorescent staining techniques were used for a systematic examination of methods used to cryopreserve microbial cell banks. The aim of cryopreservation here is to ensure subsequent reproducible fermentation performance rather than just post thaw viability. Bacillus licheniformis cell physiology post-thaw is dependent on the cryopreservant (either Tween 80, glycerol or dimethyl sulphoxide) and whilst this had a profound effect on the length of the lag phase, during subsequent 5 l fed-batch fermentations, it had little effect on maximum specific growth rate, final biomass concentration or α-amylase activity. Tween 80 not only protected the cells during freezing but also helped them recover post-thaw resulting in shorter process times.

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Abbreviations

F :

Feed rate (l h−1)

S :

Substrate concentration in the feed solution (g l−1)

μ :

Desired specific growth rate (h−1)

Y xs :

Maximum biomass yield with a limiting substrate (g/g)

X 0 :

Initial amount of cells at the start of feeding (g)

m :

Maintenance coefficient (gg−1 h−1)

t :

Time after feeding commences (h)

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Acknowledgements

Fluorochromes were provided by Molecular Probes/Invitrogen (Oregon, USA) and the authors thank William Godfrey (Invitrogen Corporation) for his helpful discussions. The authors would like to acknowledge the financial support of the Biotechnology and Biological Sciences Research Council (UK) and Novozymes A/S (Denmark) for this study.

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Authors and Affiliations

  1. School of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK

    Nichola H. Hancocks & Colin R. Thomas

  2. Novozymes A/S, Smørmosevej 25 (2JS.12), 2880, Bagsværd, Denmark

    Stuart M. Stocks

  3. Department of Chemical Engineering, Centre for Biological Engineering, Loughborough University, Loughborough, Leicestershire, LE11 3TU, UK

    Christopher J. Hewitt

Authors
  1. Nichola H. Hancocks
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  2. Colin R. Thomas
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  3. Stuart M. Stocks
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  4. Christopher J. Hewitt
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Correspondence to Christopher J. Hewitt.

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Hancocks, N.H., Thomas, C.R., Stocks, S.M. et al. An investigation into the preservation of microbial cell banks for α-amylase production during 5 l fed-batch Bacillus licheniformis fermentations. Biotechnol Lett 32, 1405–1412 (2010). https://doi.org/10.1007/s10529-010-0308-7

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  • DOI: https://doi.org/10.1007/s10529-010-0308-7

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