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Mass Spectrometry Feedback Control for Synthesis of Polyhydroxyalkanoate Granule Microstructures in Ralstonia eutropha

  1. Erik N. Pederson,
  2. Friedrich Srienc*

Article first published online: 23 MAR 2004

DOI: 10.1002/mabi.200300137

Issue

Macromolecular Bioscience

Macromolecular Bioscience

Special Issue: Highlights from the First IUPAC International Conference on Bio-based Polymers (ICBP 2003) in Saitama, Japan

Volume 4, Issue 3, pages 243–254, March 15, 2004

Additional Information

How to Cite

Pederson, E. N. and Srienc, F. (2004), Mass Spectrometry Feedback Control for Synthesis of Polyhydroxyalkanoate Granule Microstructures in Ralstonia eutropha. Macromol. Biosci., 4: 243–254. doi: 10.1002/mabi.200300137

Author Information

  1. Department of Chemical Engineering and Materials Science, and BioTechnology Institute, University of Minnesota, Minneapolis/St. Paul, MN 55455/55108, 240 Gortner Lab, 1479 Gortner Ave., St. Paul, MN 55108, USA

*Department of Chemical Engineering and Materials Science, and BioTechnology Institute, University of Minnesota, Minneapolis/St. Paul, MN 55455/55108, 240 Gortner Lab, 1479 Gortner Ave., St. Paul, MN 55108, USA. Fax: (+1) 612-625-1700

Publication History

  1. Issue published online: 23 MAR 2004
  2. Article first published online: 23 MAR 2004
  3. Manuscript Accepted: 23 JAN 2004
  4. Manuscript Revised: 20 JAN 2004
  5. Manuscript Received: 23 DEC 2003

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Keywords:

  • biopolymers;
  • core-shell polymers;
  • mass spectrometry;
  • polyesters;
  • TEM

Abstract

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Summary: Polyhydroxyalkanoate (PHA) granules with core-shell layered microstructure were synthesized in Ralstonia eutropha using periodic feeding of valeric acid into a growth medium containing excess fructose. The O2 consumption and CO2 evolution rates, determined by off-gas mass spectrometry, have been used as sensitive measures to indicate the type of nutrients utilized by R. eutropha during PHA synthesis. Domains of poly-3-hydroxybutyrate (PHB) were formed during polymer storage conditions when only fructose was present. Feeding of valeric acid (pentanoic acid) resulted in the synthesis of hydroxyvalerate (HV) monomers, forming a poly-3-hydroxybutyrate-co-valerate (PHBV) copolymer. The synthesis of desired polymer microstructures was monitored and controlled using online mass spectrometry (MS). The respiratory quotient (RQ) was unique to the type of polymer being synthesized due to increased O2 consumption during PHBV synthesis. MS data was used as the control signal for nutrient feeding strategies in the bioreactor. The core-shell structures synthesized were verified in cells using transmission electron microscopy after thin sectioning and staining with RuO4. It was demonstrated that the synthesis of core-shell microstructures can be precisely controlled utilizing a MS feedback control system.

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