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Biotechnol Lett. 2003 Nov;25(21):1795-800.

On-line monitoring of the methanol concentration in Pichia pastoris cultures producing an heterologous lipase by sequential injection analysis.

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  • 1Departament d'Enginyeria Química, ETSE, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.


An automated sequential injection analysis (SIA) system using stop-flow technique was developed to determine methanol concentration by means of the enzymatic reactions of alcohol oxidase and peroxidase. Its application as an on-line device for monitoring Pichia pastoris fermentations producing an heterologous protein was demonstrated. Linear response, observed up to 2 g l(-1), was reached by including a dilution chamber in the SIA manifold. The sampling frequency was 7 analyses per hour with a relative standard deviation lower than 4%.

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