A Simple Route for Purifying Extracellular Poly(3-hydroxybutyrate)-depolymerase from Penicillium pinophilum
Author(s) -
Elpiniki Panagiotidou,
Constantinos Konidaris,
Apostolos Baklavaridis,
Ioannis Zuburtikudis,
Dimitris S. Achilias,
Paraskevi Mitlianga
Publication year - 2014
Publication title -
enzyme research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.439
H-Index - 39
eISSN - 2090-0406
pISSN - 2090-0414
DOI - 10.1155/2014/159809
Subject(s) - enzyme , extracellular , chromatography , polyacrylamide gel electrophoresis , isolation (microbiology) , enzyme assay , chemistry , penicillium , biochemistry , biology , microbiology and biotechnology , food science
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.
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