On-line monitoring of Phaffia rhodozyma fed-batch process with in situ dispersive raman spectroscopy

Authors

  • Christopher Cannizzaro,

    1. Laboratory of Chemical and Biochemical Engineering, Department of Chemistry, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland; telephone: +41-21-693-3191; fax: +41-21-693-3680
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  • Martin Rhiel,

    1. Laboratory of Chemical and Biochemical Engineering, Department of Chemistry, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland; telephone: +41-21-693-3191; fax: +41-21-693-3680
    Current affiliation:
    1. Process Development, Cytos Biotechnology AG, Zürich-Schlieren, Switzerland
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  • Ian Marison,

    1. Laboratory of Chemical and Biochemical Engineering, Department of Chemistry, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland; telephone: +41-21-693-3191; fax: +41-21-693-3680
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  • Urs von Stockar

    Corresponding author
    1. Laboratory of Chemical and Biochemical Engineering, Department of Chemistry, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland; telephone: +41-21-693-3191; fax: +41-21-693-3680
    • Laboratory of Chemical and Biochemical Engineering, Department of Chemistry, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland; telephone: +41-21-693-3191; fax: +41-21-693-3680
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Abstract

Since the yeast Phaffia rhodozyma was first described some 35 years ago, there has been significant interest in the development of commercial processes to exploit its ability to produce carotenoids (∼80% astaxanthin). However, the optimal conditions for carotenoid production are not well understood. A key limitation has been the lack of an appropriate sensor for on-line carotenoid quantification. In this study, an in situ Raman spectroscopy probe was used to monitor intracellular carotenoid production for three consecutive P. rhodozyma fed-batch experiments. Raman spectroscopy is particularly well suited to the study of carotenoids due to a resonance effect, which greatly enhances the intensity of the three fundamental carotenoid bands, ν1 (1513 cm−1, Cmath imageC stretch), ν2 (1154 cm−1, C—C stretch), and ν3 (1003 cm−1, CH3 rock). For all three cultures, the peak height of these bands was linearly correlated with intracellular carotenoid content (1 to 45 mg/L) to a precision of better than 5%, and the correlation from one experiment was directly applicable to others. ©2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 668–680, 2003.

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