Control of continuous polyhydroxybutyrate synthesis using calorimetry and flow cytometry

Thomas Maskow, Susann Müller, Andreas Lösche, Hauke Harms, Richard Bernard Kemp

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21 Citations (SciVal)

Abstract

The substrate-carbon flow can be controlled in continuous bioreactor cultures by the medium composition, for example, by the C/N ratio. The carbon distribution is optimal when a maximum fraction flows into the desired product and the residual is just sufficient to compensate for the dilution of the microbial catalyst. Undershooting of the latter condition is reflected immediately by changes in the Gibbs energy dissipation and cellular states. Two calorimetric measurement principles were applied to optimize the continuous synthesis of polyhydroxybutyrate (PHB) by Variovorax paradoxus DSM4065 during growth with constantly increasing supply rates of fructose or toxic phenol. Firstly, the changed slope of the heat production rate in a complete heat balanced bioreactor (CHB) indicated optimum carbon channeling into PHB. The extent of the alteration depended directly on the toxic properties of the substrate. Secondly, a flow through calorimeter was connected with the bioreactor as a ‘‘measurement loop.’’ The optimum substrate carbon distribution was indicated by a sudden change in the heat production rate independent of substrate toxicity. The sudden change was explained mathematically and exploited for the long-term control of phenol conversion into PHB. LASER flow cytometry measurements distinguished between subpopulations with completely different PHB-content. Populations grown on fructose preserved a constant ratio of two subpopulations with double and quadruple sets of DNA. Cells grown on phenol comprised a third subpopulation with a single DNA set. Rising phenol concentrations caused this subpopulation to increase. It may thus be considered as an indicator of chemostress.
Original languageEnglish
Pages (from-to)541-552
Number of pages12
JournalBiotechnology and Bioengineering
Volume93
Issue number3
DOIs
Publication statusPublished - 21 Oct 2005

Keywords

  • chemostat
  • calorimetry flow cytometry
  • polyhydroxbutyrate (PHB)
  • phenol
  • Variovorax paradoxus DSM 4065

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