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Chemometric analysis of in-line multi-wavelength fluorescence measurements obtained during cultivations with a lipase producing Aspergillus oryzae strain
Authors:Haack Martin B  Lantz Anna Eliasson  Mortensen Peter P  Olsson Lisbeth
Institution:1. Center for Microbial Biotechnology, BioCentrum‐DTU, Technical University of Denmark, S?ltofts Plads, Building 223, DK‐2800 Kgs. Lyngby, Denmark;2. telephone: +45‐4525‐2677;3. fax: +45‐4588‐4148;4. Process Support Laboratories, Novozymes A/S, Hallas Alle AXS.02, DK‐4400 Kalundborg, Denmark
Abstract:The filamentous fungus, Aspergillus oryzae, was cultivated in batch and fed-batch cultivations in order to investigate the use of multi-wavelength fluorescence for monitoring course of events during filamentous fungi cultivations. The A. oryzae strain applied expressed a fungal lipase from Thermomyces lanuginosus. Spectra of multi-wavelength fluorescence were collected every 5 min with the BioView system (DELTA, Denmark) and both explorative and predictive models, correlating the fluorescence data with cell mass and lipase activity, were built. During the cultivations, A. oryzae displayed dispersed hyphal growth and under these conditions no fouling of the multi-wavelength fluorescence sensor was observed. The scores of a parallel factor analysis (PARAFAC) model, based on the fluorescence spectra, gave clear evidence of, for example, the on-set of the feeding phase. The predictive models, estimating the cell mass, showed correlations between 0.73 and 0.97 with root mean square error of cross validation (RMSECV) values between 1.48 and 0.77 g . kg(-1). A model estimating the lipase activity was also constructed for the fed-batch cultivations with a correlation of 0.93. The results presented here clearly show that multi-wavelength fluorescence is a useful tool for monitoring fed-batch cultivations of filamentous fungi.
Keywords:PAT  three‐way analysis  multivariate data analysis  cell mass  PLS
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