Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East India
The extracellular dextransucrase produced from Pediococcus pentosaceus, a new isolate from the soil in Assam, India, was purified and characterized. The enzyme activity of cell-free supernatant was 3.4 U/mL and specific activity was 0.6 U/mg. The crude enzyme was purified by a single-step fractionat...
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| Format: | Article |
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University of Zagreb Faculty of Food Technology and Biotechnology
2011-01-01
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| Series: | Food Technology and Biotechnology |
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| Online Access: | http://hrcak.srce.hr/file/105755 |
| _version_ | 1797429336348819456 |
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| author | Damini Kothari Seema Patel Arun Goyal |
| author_facet | Damini Kothari Seema Patel Arun Goyal |
| author_sort | Damini Kothari |
| collection | DOAJ |
| description | The extracellular dextransucrase produced from Pediococcus pentosaceus, a new isolate from the soil in Assam, India, was purified and characterized. The enzyme activity of cell-free supernatant was 3.4 U/mL and specific activity was 0.6 U/mg. The crude enzyme was purified by a single-step fractionation using polyethylene glycols of different molecular mass. The specific activity achieved was 18 U/mg with 31-fold purification by PEG 400 and 26 U/mg with 45-fold purification by PEG 1500. The molecular mass of dextransucrase determined by non-denaturing SDS-PAGE was approx. 180 kDa. The dextran formation activity of the enzyme was confirmed by activity staining. Optimum conditions for dextransucrase activity were: pH=5.4, reaction temperature 30 °C, 5 % sucrose and 20 mM sodium acetate buffer. A concentration of 1 mM MgCl2 and 6 mM CaCl2 enhanced dextransucrase activity by 5 and 150 %, respectively. The chaotropic agent urea (7 M) and chelating agent EDTA (1 mM) resulted in the residual enzyme activity of 98 and 80 %, respectively. The organic solvents such as ethanol (50 %), DMSO (90 %), acetone (50 %) and acetonitrile (20 %) decreased the dextransucrase activity by 80, 91, 94 and 80 %, respectively. |
| first_indexed | 2024-03-09T09:12:44Z |
| format | Article |
| id | doaj.art-34438a0608e749bfb0310110d5153608 |
| institution | Directory Open Access Journal |
| issn | 1330-9862 1334-2606 |
| language | English |
| last_indexed | 2024-03-09T09:12:44Z |
| publishDate | 2011-01-01 |
| publisher | University of Zagreb Faculty of Food Technology and Biotechnology |
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| series | Food Technology and Biotechnology |
| spelling | doaj.art-34438a0608e749bfb0310110d51536082023-12-02T08:19:41ZengUniversity of Zagreb Faculty of Food Technology and BiotechnologyFood Technology and Biotechnology1330-98621334-26062011-01-01493297303Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East IndiaDamini Kothari0Seema Patel1Arun Goyal2Department of Biotechnology, Indian Institute of Technology Guwahati, 781039 Guwahati, Assam, IndiaDepartment of Biotechnology, Indian Institute of Technology Guwahati, 781039 Guwahati, Assam, IndiaDepartment of Biotechnology, Indian Institute of Technology Guwahati, 781039 Guwahati, Assam, IndiaThe extracellular dextransucrase produced from Pediococcus pentosaceus, a new isolate from the soil in Assam, India, was purified and characterized. The enzyme activity of cell-free supernatant was 3.4 U/mL and specific activity was 0.6 U/mg. The crude enzyme was purified by a single-step fractionation using polyethylene glycols of different molecular mass. The specific activity achieved was 18 U/mg with 31-fold purification by PEG 400 and 26 U/mg with 45-fold purification by PEG 1500. The molecular mass of dextransucrase determined by non-denaturing SDS-PAGE was approx. 180 kDa. The dextran formation activity of the enzyme was confirmed by activity staining. Optimum conditions for dextransucrase activity were: pH=5.4, reaction temperature 30 °C, 5 % sucrose and 20 mM sodium acetate buffer. A concentration of 1 mM MgCl2 and 6 mM CaCl2 enhanced dextransucrase activity by 5 and 150 %, respectively. The chaotropic agent urea (7 M) and chelating agent EDTA (1 mM) resulted in the residual enzyme activity of 98 and 80 %, respectively. The organic solvents such as ethanol (50 %), DMSO (90 %), acetone (50 %) and acetonitrile (20 %) decreased the dextransucrase activity by 80, 91, 94 and 80 %, respectively.http://hrcak.srce.hr/file/105755dextransucrasepolyethylene glycolSDS-PAGEactivity stainingPediococcus pentosaceus |
| spellingShingle | Damini Kothari Seema Patel Arun Goyal Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East India Food Technology and Biotechnology dextransucrase polyethylene glycol SDS-PAGE activity staining Pediococcus pentosaceus |
| title | Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East India |
| title_full | Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East India |
| title_fullStr | Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East India |
| title_full_unstemmed | Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East India |
| title_short | Purification and Characterization of an Extracellular Dextransucrase from Pediococcus pentosaceus Isolated from the Soil of North East India |
| title_sort | purification and characterization of an extracellular dextransucrase from pediococcus pentosaceus isolated from the soil of north east india |
| topic | dextransucrase polyethylene glycol SDS-PAGE activity staining Pediococcus pentosaceus |
| url | http://hrcak.srce.hr/file/105755 |
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