title: A study on the stability and enzymatic activity of yeast alcohol dehydrogenase in presence of the self-assembling block copolymer Poloxamer 407.
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.
authors.affiliation: School of Biosciences and Biotechnology, University of Camerino, Via Gentile III da Varano, Camerino, Italy. stefania.pucciarelli@unicam.it
authors.forename: Stefania
authors.initials: S
authors.lastname: Pucciarelli
authors.forename: Giulia
authors.initials: G
authors.lastname: Bonacucina
authors.forename: Franco
authors.initials: F
authors.lastname: Bernabucci
authors.forename: Marco
authors.initials: M
authors.lastname: Cespi
authors.forename: Giovanna
authors.initials: G
authors.lastname: Mencarelli
authors.forename: Giusi Serena
authors.initials: GS
authors.lastname: De Fronzo
authors.forename: Paolo
authors.initials: P
authors.lastname: Natalini
authors.forename: Giovanni Filippo
authors.initials: GF
authors.lastname: Palmieri
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D020442:Poloxamer / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism; D013696:Temperature
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D020442:Poloxamer; D000426:Alcohol Dehydrogenase