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The Challenging Measurement of Protein in Complex Biomass-Derived Samples

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Abstract

Measurement of the protein content in samples from production of lignocellulosic bioethanol is an important tool when studying the adsorption of cellulases. Several methods have been used for this, and after reviewing the literature, we concluded that one of the most promising assays for simple and fast protein measurement on this type of samples was the ninhydrin assay. This method has also been used widely for this purpose, but with two different methods for protein hydrolysis prior to the assay—alkaline or acidic hydrolysis. In samples containing glucose or ethanol, there was significant interference from these compounds when using acid hydrolysis, which was not the case when using the alkaline hydrolysis. We evaluated the interference from glucose, cellulose, xylose, xylan, lignin and ethanol on protein determination of BSA, Accellerase® 1500 and Cellic® CTec2. The experiments demonstrated that the presence of cellulose, lignin and glucose (above 50 g/kg) could significantly affect the results of the assay. Comparison of analyses performed with the ninhydrin assay and with a CN analyser revealed that there was good agreement between these two analytical methods, but care has to be taken when applying the ninhydrin assay. If used correctly, the ninhydrin assay can be used as a fast method to evaluate the adsorption of cellulases to lignin.

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Acknowledgments

We wish to thank Kathrine B. Hansen and Tanya Jensen for technical assistance on reference analyses of nitrogen on elemental analyser, Georg Ørnskov Rønsch for his help on the experimental design of 52 experiment, Kit Kellebjerg Mogensen and Martin Dan Jeppesen for valuable discussions and Vibeke Ørsted Bonde Nielsen for technical assistance on ninhydrin assay on the reference samples. The industrial Ph.D. project is financially supported by the Danish Agency for Science, Technology, and Innovation, grant no. 09–053694.

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Authors and Affiliations

  1. DONG Energy A/S, Kraftværksvej 53, 7000, Fredericia, Denmark

    Mai Østergaard Haven

  2. Department of Geosciences and Natural Resource Management, University of Copenhagen, Rolighedsvej 23, 1958, Frederiksberg C, Denmark

    Mai Østergaard Haven & Henning Jørgensen

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  1. Mai Østergaard Haven
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  2. Henning Jørgensen
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Correspondence to Mai Østergaard Haven.

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Haven, M.Ø., Jørgensen, H. The Challenging Measurement of Protein in Complex Biomass-Derived Samples. Appl Biochem Biotechnol 172, 87–101 (2014). https://doi.org/10.1007/s12010-013-0466-x

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  • DOI: https://doi.org/10.1007/s12010-013-0466-x

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