Production and Characterization of Tannase By Bacillus subtilis in Solid State Fermentation of Corn Leaves

Shakir, Hafiz Abdullah; Khan, Muhammad; Irfan, Muhammad; Ali, Shaukat; Yousaf, Muhammad Abrar; Javed, Iqra; Qazi, Javed Iqbal; Bukhari, Syed Shahid Imran

doi:10.30491/jabr.2021.265181.1334

Production and Characterization of Tannase By Bacillus subtilis in Solid State Fermentation of Corn Leaves

Document Type : Original Article

Authors

Hafiz Abdullah Shakir ¹

Muhammad Khan ¹

Muhammad Irfan ²

Shaukat Ali ³

Muhammad Abrar Yousaf ¹

Iqra Javed ¹

Javed Iqbal Qazi ¹

Syed Shahid Imran Bukhari ³

¹ Institute of Zoology, Faculty of Life Sciences, University of the Punjab, Lahore, Pakistan

² Department of Biotechnology, University of Sargodha, Sargodha, Pakistan

³ Department of Zoology, Faculty of Sciences, Government College University, Lahore, Pakistan

10.30491/jabr.2021.265181.1334

Abstract

Introduction: Tannase (tannin acyl hydrolase EC3.1.1.20) is an industrially important enzyme with extensive applications. The current study aimed to optimize the tannase production employing corn leaves as substrate and characterize tannase activity.
Materials and Methods: Tannase producing bacterial strains were isolated from Catla catla fish gut. The highest enzyme-producing bacterial strain was identified as Bacillus subtilis using 16S rDNA sequencing.
Results: Fermentation parameters and additional medium components were optimized with the application of one variable at a time and enhanced tannase was obtained with 50% substrate moisture, acetate buffer (pH 4.0) as enzyme extraction medium with 2 ml volume, 45 °C incubation temperature, pH 5, 2% inoculum size, 24 h incubation time, 150 rpm agitation, large-sized substrate particles (4.0 mm), enzyme extraction without centrifugation and medium components (MgSO₄, 4% tannic acid and yeast extract). The central composite design was employed to optimize the concentrations of optimal medium components, which were found as 4.0% tannic acid, 0.5% MgSO₄ and 1.5% yeast extract for the highest tannase production (211.97±0.08 U/ml). Tannase characterization revealed the maximum tannase activity at pH 8, 30 °C (with 30 min incubation period and 0.35% substrate concentration).
Conclusions: The results of the present study revealed the potential of utilization of agricultural resources (corn leaves) as a low-cost substrate to reduce the production cost of tannase.

Keywords

Bacillus subtilis

Tannase

Production

Central Composite Design

Characterization