ISSN: 0973-7510

E-ISSN: 2581-690X

Research Article | Open Access
Nishanthini Thangavelu, Priyanka Hugar and Prasanna D. Belur
Department of Chemical Engineering, National Institute of Technology Karnataka, Surathkal, Srinivasanagar, Mangalore, Karnataka, India.
Article Number: 7819 | © The Author(s). 2022
J Pure Appl Microbiol. 2022;16(4):2557-2567. https://doi.org/10.22207/JPAM.16.4.22
Received: 06 May 2022 | Accepted: 03 September 2022 | Published online: 21 October 2022
Issue online: December 2022
Abstract

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.

Keywords

Geotrichum cucujoidarum, Isolation, Production, Tannase

Article Metrics

Article View: 81

Share This Article

© The Author(s) 2022. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.