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A Study on the Stability and Enzymatic Activity of Yeast Alcohol Dehydrogenase in Presence of the Self-Assembling Block Copolymer Poloxamer 407

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Abstract

Yeast alcohol dehydrogenase (ADH) is an enzyme widely studied for biotechnological applications due to its involvement in fermentation industry, and various attempts to improve its catalytic properties and its thermal stability have been carried out. In this paper, the influence of a block copolymer (Poloxamer 407) on ADH enzymatic activity and thermal behaviour has been studied in order to get new insights about the use of poloxamers in formulation of sustained release systems for therapeutic proteins. Poloxamer 407 has the ability to form micelles and gel due to its self-assembling and thermoresponsive properties. The effect of the copolymer towards thermal stress and pH changes, which often reduce enzymes activity it has been investigated by means of enzymatic assays and differential scanning calorimetry. Results showed that at pH 9.1 and 7.3, the Poloxamer in the form of unimeric, micellar and gel state is able to effectively preserve the enzyme from thermoinactivation. In addition by calorimetric data Poloxamer 407 has showed an effect in preserving ADH from aggregation at pH 7.3. In conclusion, Poloxamer 407 seems to be very effective in protecting ADH from stress related events, like alkaline inactivation and aggregation.

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Authors and Affiliations

  1. School of Biosciences and Biotechnology, University of Camerino, Via Gentile III da Varano, Camerino, Italy

    Stefania Pucciarelli, Franco Bernabucci, Giusi Serena De Fronzo & Paolo Natalini

  2. School of Pharmacy, University of Camerino, Via S. Agostino 1, Camerino, Italy

    Giulia Bonacucina, Marco Cespi, Giovanna Mencarelli & Giovanni Filippo Palmieri

Authors
  1. Stefania Pucciarelli
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  2. Giulia Bonacucina
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  3. Franco Bernabucci
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  4. Marco Cespi
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  5. Giovanna Mencarelli
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  6. Giusi Serena De Fronzo
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  7. Paolo Natalini
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  8. Giovanni Filippo Palmieri
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Correspondence to Stefania Pucciarelli.

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Pucciarelli, S., Bonacucina, G., Bernabucci, F. et al. A Study on the Stability and Enzymatic Activity of Yeast Alcohol Dehydrogenase in Presence of the Self-Assembling Block Copolymer Poloxamer 407. Appl Biochem Biotechnol 167, 298–313 (2012). https://doi.org/10.1007/s12010-012-9692-x

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  • DOI: https://doi.org/10.1007/s12010-012-9692-x

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