Isolation and screening of fungal isolates from bambara (Vigna subterranea) nuts for tannase production

Abstract

Tannase (Tannin acyl hydrolase, EC 3.1.1.20) is an enzyme produced in the presence of tannic acid by various filamentous fungi. They are produced principally by fungi of the genus Aspergillus and Penicillium. The enzyme is used in the food and beverage industry as a clarifying agent for wines, beers and fruit juices. In Africa, billions of dollars are expended yearly on the importation of commercial enzymes for the food and pharmaceutical industries and this increases the cost of production and the finished goods. This study was carried out to isolate tannase producing fungal species using Bambara nuts as a substrate in a bid to finding alternatives to the importation of tannase. Fresh Bambara nuts were collected from different locations in Nigeria. They were cleaned, sorted and intermittently moistened with water to encourage fungal growth for fourteen days. The different fungi obtained after fourteen days were inoculated onto Potato Dextrose Agar plates and incubated at 25°C for five days. Subculturing of fungal isolates was carried out to obtain pure cultures of isolates. Tannilytic activity (hydrolysis of tannin) of isolates was assessed by inoculating them in media containing tannin. The plates were incubated at 25°C for 2-5 days after which the plates were observed and zones of hydrolysis measured. A total of eighteen isolates were obtained. They were all members of the Aspergillus genus. 56% (10) of the isolates were able to degrade tannin acid with mean zone of hydrolysis of 39mm ±23.7 mm (Range 10-70mm). This study established members of the Aspergillus genus isolated from Bambara nuts as viable fungi for application in the production of tannase. This study adds to existing reports on fungal production of tannase.

Keywords: Tannase; Fungal isolates; Aspergillus; Bambara nuts; Vigna subterranea