Abstract
A method for assay of microbial tannase (Tannin acyl hydrolase) based on the formation of chromogen between gallic acid and rhodanine is reported. Maximum Tannase production occurred in the culture broth containing 1-2% (w/v) tannic acid and 0.05 – 0.1% (w/v) glucose. The pH, incubation period, temperature and Glucose concentration optima of Tannase production was found at 5.5, 36 h, 35°C and 0.5% respectively. These properties make the enzyme suitable for pollution control and bioprocess industry. This assay is very simple, reproducible, and very convenient, and with it Tannase activity can be measured in relation to the growth of the organism. Aspergillus niger exhibited higher enzyme activity showing about 65 mole percent conversion respectively after a 36 h incubation period. The assay is complete in a short time, very convenient and reproducible.
Highlights
Tannin acyl hydrolase commonly called tannase is produced by a number of microorganisms like fungi (Aspergillus, Penicillium, Rhizopus sp), yeast (Candida sp), and bacteria (Bacillus sp)[1,2]
The major commercial application of this enzyme is in the hydrolysis of gallotannin to Gallic acid, is an intermediate required for the synthesis of an antifolic antibacterial drug trimethoprim[3]
Studies on the tannase production from Aspergillus niger was carried out and the results were given in the Tables and Figs
Summary
Tannin acyl hydrolase commonly called tannase is produced by a number of microorganisms like fungi (Aspergillus, Penicillium, Rhizopus sp), yeast (Candida sp), and bacteria (Bacillus sp)[1,2]. Inoue and Hagerman, have described a method for the determination of gallotannins, which involves the formation of a chromogen between gallic acid obtained by the acid hydrolysis of gallotannis and rhodanine[9] This was later adapted by[10], for the assay of tannase in a ruminal bacterium. 50 mL gallotannin containing potato dextrose medium was taken, each one is transferred to respective 250 mL conical flask and sterilized These flasks were inoculated aseptically with 2 mL of spore suspension prepared from the culture slants. Enzyme productin was carried out in 250-mL Erlenmeyer flasks containing 50-mL 2% tannic acid in potato dextrose broth (pH 5.5) medium was taken in 100 mL conical flasks It was sterilized and inoculated with 2% (v/v) inoculum was added and incubated at 35 °C for 36 h in a rotary shaker (200-rpm). One unit of the tannase was defined as the amount of enzyme, which is able to hydrolyse 1μ mole of ester linkage of tannic acid in 1 min at specific condition
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