Optimization of production and reaction conditions of polygalacturonase from Byssochlamys fulva

Gupta, Reena and Kalpana, (2011) Optimization of production and reaction conditions of polygalacturonase from Byssochlamys fulva. Acta Microbiologica et Immunologica Hungarica, 58 (4). pp. 339-349. ISSN 1217-8950

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Abstract

In the present study, the optimization of production and reaction conditions of polygalacturonase produced by a fungus Byssochlamys fulva MTCC 505 was achieved. The production of polygalacturonase with a considerable activity of 1.28 IU/ml was found when the culture was shaken at 30°C for 5 days in 100 ml of medium containing (w/v) 10 g/l pectin, 2 g/l NaNO3, 1 g/l KH2PO4, 0.5 g/l KCl, 0.5 g/l MgSO4. 7H2O, 0.001 g/l FeSO4. 7H2O, 0.001 g/l CaCl2. The best carbon and nitrogen source for this enzyme were pectin (1%) and Ca(NO3)2 (0.1%), respectively. The enzyme gave maximum activity at incubation time of 72 h, temperature of 30°C and pH 4.5. During the optimization of reaction conditions, the enzyme showed maximum activity in sodium citrate buffer (50 mM) of pH 5.5 at 50°C reaction temperature for 15 minutes of incubation. The enzyme showed greater affinity for polygalacturonic acid as substrate (0.5%). Km and Vmax values were 0.15 mg/ml and 4.58 μmol/ml/min. The effect of various phenolics, thiols, protein inhibitors and metal ions on the enzyme activity was investigated. The enzyme was quite stable at 4°C and 30°C. At 40°C the half life of the enzyme was 6 h and at 60°C it was 2 h.

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