Improvement of Endoglucanase Activity in Penicillium oxalicum ID10-T065 Mutated by Ultra Violet Irradiation and Ethidium Bromide

Asnany Caniago, Wibowo Mangunwardoyo, Sukma Nuswantara, Puspita Lisdiyanti

Abstract


Penicillium sp. is known as filamentous fungi that produce complete cellulase. Cellulase. This study aims to improve endoglucanase activity of Penicillium oxalicum ID010-T065 by mutated with ultra violet irradiation (with dose of 0.1 J/cm2, 15 cm), ethidium bromide (10 µg/mL, 1 hour) and combination of both mutagens. The endoglucanase activity of all mutants was higher than that of the wild type (1.03 U/mL). Mutant UVEB-42 exposed to combine mutation showed the highest endoglucanase activity (2.76 U/mL) with a 2.70 fold increase. Mutant EB-45 (1.83 U/mL) exposed to ethidium bromide solution showed a 1.8 fold increase. Mutant UV-13 (1.72 U/mL) exposed to UV irradiation for 3 minutes showed a 1.7 fold increase. All mutants have optimum endoglucanase activity at 50 °C. Mutant UVEB-53 showed the highest thermostability by retaining 86 % of endoglucanase activity at 90 °C. The gene analysis of the endoglucanase I gene showed 3 bases mutated at mutant UV-13 and UVEB-53 that changed proline to serine. Mutant EB-45 showed 4 bases mutated that changed valine to glysine and proline to serine. Two bases mutated at Mutant UVEB-53 changed proline to serine. Bases mutated in eg1 gene could influenced the enhance of enzym activity in mutant.

Keywords


endoglucanase; mutation; ultra violet; ethidium bromide; Penicillium

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References


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DOI: http://dx.doi.org/10.14203/ann.bogor.2015.v19.n2.27-38

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