A Simple Route for Purifying Extracellular Poly(3-hydroxybutyrate)-depolymerase from Penicillium pinophilum

Elpiniki Panagiotidou; Constantinos Konidaris; Apostolos Baklavaridis; Ioannis Zuburtikudis; Dimitris Achilias; Paraskevi Mitlianga

doi:10.1155/2014/159809

A Simple Route for Purifying Extracellular Poly(3-hydroxybutyrate)-depolymerase from Penicillium pinophilum

Enzyme Res. 2014:2014:159809. doi: 10.1155/2014/159809. Epub 2014 Sep 23.

Authors

Elpiniki Panagiotidou¹, Constantinos Konidaris², Apostolos Baklavaridis³, Ioannis Zuburtikudis⁴, Dimitris Achilias⁵, Paraskevi Mitlianga⁶

Affiliations

¹ Department of Mechanical and Industrial Design Engineering, TEI of Western Macedonia, 50100 Kozani, Greece ; Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
² Laboratory of Biotechnology, Department of Biological Applications and Technologies, University of Ioannina, 45100 Ioannina, Greece ; Laboratory of Biochemistry, Department of Biological Applications and Technologies, University of Ioannina, 45100 Ioannina, Greece.
³ Department of Mechanical and Industrial Design Engineering, TEI of Western Macedonia, 50100 Kozani, Greece.
⁴ Department of Mechanical and Industrial Design Engineering, TEI of Western Macedonia, 50100 Kozani, Greece ; Department of Chemical and Petroleum Engineering, United Arab Emirates University, Al Ain, UAE.
⁵ Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
⁶ Depertment of Agricultural Technology, School of Agriculture Technology, Food Technology and Nutrition, TEI of Western Macedonia, Terma Kontopoulou, 53100 Florina, Greece.

Abstract

This work proposes the purification of an active and efficient enzyme, extracellular poly(3-hydroxybutyrate) (PHB)-depolymerase, suitable for industrial applications. This is achieved by the application of an easy, fast, and cheap route, skipping the chromatography step. Chromatography with one or two columns is a common step in the purification procedure, which however renders the isolation of the enzyme a time consuming and an expensive process. A strain of the fungus Penicillium pinophilum (ATCC 9644) is used for the isolation of extracellular PHB-depolymerase. The molecular weight of the purified enzyme is about 35 kDa and is estimated by gel electrophoresis (SDS-PAGE, 12% polyacrylamide). The enzymatic activity of the isolated enzyme is determined to be 3.56-fold similar to that found by other researchers that have used chromatography for the isolation. The as-isolated enzyme disintegrates the poly(3-hydroxybutyrate) (PHB) films successfully, as it is demonstrated by the biodegradation test results provided here.