Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarum
With an aim to isolate a tannase positive organism, the microbial mat growing on the stored areca extract leachate surface was screened. Once the tannase positive organism was isolated, it was identified by ITS/18S rRNA gene sequencing. Further, the enzyme was purified and examined for its biochemic...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Journal of Pure and Applied Microbiology
2022-12-01
|
| Series: | Journal of Pure and Applied Microbiology |
| Subjects: | |
| Online Access: | https://microbiologyjournal.org/production-purification-and-characterization-of-extracellular-tannase-from-a-newly-isolated-yeast-geotrichum-cucujoidarum/ |
| _version_ | 1797950953255600128 |
|---|---|
| author | Nishanthini Thangavelu Priyanka Hugar Prasanna D. Belur |
| author_facet | Nishanthini Thangavelu Priyanka Hugar Prasanna D. Belur |
| author_sort | Nishanthini Thangavelu |
| collection | DOAJ |
| description | With an aim to isolate a tannase positive organism, the microbial mat growing on the stored areca extract leachate surface was screened. Once the tannase positive organism was isolated, it was identified by ITS/18S rRNA gene sequencing. Further, the enzyme was purified and examined for its biochemical properties. A potent extracellular tannase-producing yeast was isolated and was identified as Geotrichum cucujoidarum. After the shake flask studies, the enzyme activity of 4.42 U/ml and specific activity of 29.86 U/mg were achieved in a medium with tannic acid as an inducer. Later, ethanol (70%) precipitation followed by purification through FPLC using SEC 650 column resulted in 166.37 U/mg specific activity and a recovery of 50.54%. The purified enzyme was a monomer with a molecular weight of 63 kDa. The optimum pH and the temperature of the enzyme were found to be 5.0 and 30°C, respectively. The Michaelis-Menten constant (Km) was found to be 2.9 mM, and the turn over number (kcat) and catalytic efficiency (kcat/km) of the purified tannase were 102 S-1 and 35.17 mM-1S-1 respectively. Temperature and pH stability profiles of the enzyme, influence of various metal ions, chelators and surfactants on enzyme activity and kinetic constants of enzyme shows that the tannase produced from Geotrichum cucujoidarum is unique and is a potential candidate for further studies. |
| first_indexed | 2024-04-10T22:23:13Z |
| format | Article |
| id | doaj.art-83b0445a67974b1e8fefe62fcd5028f1 |
| institution | Directory Open Access Journal |
| issn | 0973-7510 2581-690X |
| language | English |
| last_indexed | 2024-04-10T22:23:13Z |
| publishDate | 2022-12-01 |
| publisher | Journal of Pure and Applied Microbiology |
| record_format | Article |
| series | Journal of Pure and Applied Microbiology |
| spelling | doaj.art-83b0445a67974b1e8fefe62fcd5028f12023-01-17T13:24:43ZengJournal of Pure and Applied MicrobiologyJournal of Pure and Applied Microbiology0973-75102581-690X2022-12-0116425572567https://doi.org/10.22207/JPAM.16.4.22Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarumNishanthini ThangaveluPriyanka HugarPrasanna D. Belurhttps://orcid.org/0000-0003-2908-4511With an aim to isolate a tannase positive organism, the microbial mat growing on the stored areca extract leachate surface was screened. Once the tannase positive organism was isolated, it was identified by ITS/18S rRNA gene sequencing. Further, the enzyme was purified and examined for its biochemical properties. A potent extracellular tannase-producing yeast was isolated and was identified as Geotrichum cucujoidarum. After the shake flask studies, the enzyme activity of 4.42 U/ml and specific activity of 29.86 U/mg were achieved in a medium with tannic acid as an inducer. Later, ethanol (70%) precipitation followed by purification through FPLC using SEC 650 column resulted in 166.37 U/mg specific activity and a recovery of 50.54%. The purified enzyme was a monomer with a molecular weight of 63 kDa. The optimum pH and the temperature of the enzyme were found to be 5.0 and 30°C, respectively. The Michaelis-Menten constant (Km) was found to be 2.9 mM, and the turn over number (kcat) and catalytic efficiency (kcat/km) of the purified tannase were 102 S-1 and 35.17 mM-1S-1 respectively. Temperature and pH stability profiles of the enzyme, influence of various metal ions, chelators and surfactants on enzyme activity and kinetic constants of enzyme shows that the tannase produced from Geotrichum cucujoidarum is unique and is a potential candidate for further studies.https://microbiologyjournal.org/production-purification-and-characterization-of-extracellular-tannase-from-a-newly-isolated-yeast-geotrichum-cucujoidarum/geotrichum cucujoidarumisolationproductiontannase |
| spellingShingle | Nishanthini Thangavelu Priyanka Hugar Prasanna D. Belur Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarum Journal of Pure and Applied Microbiology geotrichum cucujoidarum isolation production tannase |
| title | Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarum |
| title_full | Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarum |
| title_fullStr | Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarum |
| title_full_unstemmed | Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarum |
| title_short | Production, Purification and Characterization of Extracellular Tannase from a Newly Isolated Yeast, Geotrichum cucujoidarum |
| title_sort | production purification and characterization of extracellular tannase from a newly isolated yeast geotrichum cucujoidarum |
| topic | geotrichum cucujoidarum isolation production tannase |
| url | https://microbiologyjournal.org/production-purification-and-characterization-of-extracellular-tannase-from-a-newly-isolated-yeast-geotrichum-cucujoidarum/ |
| work_keys_str_mv | AT nishanthinithangavelu productionpurificationandcharacterizationofextracellulartannasefromanewlyisolatedyeastgeotrichumcucujoidarum AT priyankahugar productionpurificationandcharacterizationofextracellulartannasefromanewlyisolatedyeastgeotrichumcucujoidarum AT prasannadbelur productionpurificationandcharacterizationofextracellulartannasefromanewlyisolatedyeastgeotrichumcucujoidarum |