Production of laccase byBotrytis cinerea and fermentation studies with strain F226 |
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Authors: | M G Fortina A Acquati P Rossi P L Manachini C Di Gennaro |
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Affiliation: | (1) Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche, Sezione di Microbiologia Industriale, Università degli Studi di Milano, Via Celoria 2, 20133 Milano, Italy;(2) Enichem Synthesis Spa, San Donato Milanese, Milano, Italy |
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Abstract: | After induction, seven strains ofBotrytis cinerea released into the culture broth considerable amounts of laccase in a brief production time. The set-up of a suitable production process was studied with a selected strain in a 10-L fermenter. The optimum fermentation conditions were a 3% inoculum with a high degree of sporulation, a simple medium containing 20 g L–1 of glucose and 2 g L–1 of yeast extract at pH 3.5, 2 g L–1 gallic acid as inducer, added after 2 days of growth, an agitation speed of 300 rpm, an aeration rate of 1.2 vvm and a temperature of 24°C. By optimizing the culture conditions, the enzyme activity reached 28 U ml–1 in 5 days with a specific activity of 560 U mg–1 protein. The best procedure to obtain a suitable crude enzyme preparation was concentration of the supernatant medium to 10% of the initial volume by ultrafiltration, followed by a fractional precipitation with ethanol. The optimum pH and temperature for laccase activity were 5.5 and 40°C, respectively, with syringaldazine as the substrate. |
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Keywords: | laccase Botrytis cinerea scaling up optimization |
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