Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani Thom
Fusarium solani Thom produced maximum PHB depolymerase by 48 h when grown in BHM containing 0.2%, w/v PHB, pH 8.0 at 25°C. Statistical optimization studies using Plackett Burman design of PHB depolymerase production yielded maximum PHB depolymerase activity after 2 days as against 4 days in the unop...
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Wiley
2013-01-01
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| Series: | Journal of Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2013/406386 |
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| author | Srividya Shivakumar |
| author_facet | Srividya Shivakumar |
| author_sort | Srividya Shivakumar |
| collection | DOAJ |
| description | Fusarium solani Thom produced maximum PHB depolymerase by 48 h when grown in BHM containing 0.2%, w/v PHB, pH 8.0 at 25°C. Statistical optimization studies using Plackett Burman design of PHB depolymerase production yielded maximum PHB depolymerase activity after 2 days as against 4 days in the unoptimized conditions with a 2-fold increase in activity. Partial purification of the extracellular poly-β-hydroxybutyrate (PHB) depolymerase PHAZFus from F. solani Thom by two steps using ammonium sulphate (80% saturation) and affinity chromatography using concanavalin-A yielded 162.3-fold purity and 63% recovery of protein. The enzyme composed of a single polypeptide chain of 85 KDa, as determined by SDS-PAGE. The enzyme stained positive for glycoprotein by PAS staining. Optimum enzyme activity was detected at pH 7.0 and 55°C. The enzyme was stable at pH 7.0 and 55°C for 24 h with a residual activity of almost 85%. Paper chromatography revealed β-hydroxybutyrate monomer as the major end product of PHB hydrolysis. Complete inhibition of the enzyme by 1 mM HgCl2 (100%) indicated the importance of essential disulfide bonds (cystine residues) for enzyme activity or probably for maintaining the native enzyme structure. |
| format | Article |
| id | doaj-art-d5b454c8f58d47e3896c698e0c7e9465 |
| institution | OA Journals |
| issn | 2090-9063 2090-9071 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
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| series | Journal of Chemistry |
| spelling | doaj-art-d5b454c8f58d47e3896c698e0c7e94652025-08-20T02:19:40ZengWileyJournal of Chemistry2090-90632090-90712013-01-01201310.1155/2013/406386406386Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani ThomSrividya Shivakumar0Department of Microbiology, Centre for Post Graduate Studies, Jain University, 18/3, 9th Main, Jayanagar 3rd Block, Bangalore 560011, IndiaFusarium solani Thom produced maximum PHB depolymerase by 48 h when grown in BHM containing 0.2%, w/v PHB, pH 8.0 at 25°C. Statistical optimization studies using Plackett Burman design of PHB depolymerase production yielded maximum PHB depolymerase activity after 2 days as against 4 days in the unoptimized conditions with a 2-fold increase in activity. Partial purification of the extracellular poly-β-hydroxybutyrate (PHB) depolymerase PHAZFus from F. solani Thom by two steps using ammonium sulphate (80% saturation) and affinity chromatography using concanavalin-A yielded 162.3-fold purity and 63% recovery of protein. The enzyme composed of a single polypeptide chain of 85 KDa, as determined by SDS-PAGE. The enzyme stained positive for glycoprotein by PAS staining. Optimum enzyme activity was detected at pH 7.0 and 55°C. The enzyme was stable at pH 7.0 and 55°C for 24 h with a residual activity of almost 85%. Paper chromatography revealed β-hydroxybutyrate monomer as the major end product of PHB hydrolysis. Complete inhibition of the enzyme by 1 mM HgCl2 (100%) indicated the importance of essential disulfide bonds (cystine residues) for enzyme activity or probably for maintaining the native enzyme structure.http://dx.doi.org/10.1155/2013/406386 |
| spellingShingle | Srividya Shivakumar Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani Thom Journal of Chemistry |
| title | Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani Thom |
| title_full | Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani Thom |
| title_fullStr | Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani Thom |
| title_full_unstemmed | Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani Thom |
| title_short | Poly-β-hydroxybutyrate (PHB) Depolymerase from Fusarium solani Thom |
| title_sort | poly β hydroxybutyrate phb depolymerase from fusarium solani thom |
| url | http://dx.doi.org/10.1155/2013/406386 |
| work_keys_str_mv | AT srividyashivakumar polybhydroxybutyratephbdepolymerasefromfusariumsolanithom |