On-line monitoring of the methanol concentration in Pichia pastoris cultures producing an heterologous lipase by sequential injection analysis.
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%.
Surribas
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Departament d'Enginyeria Química, ETSE, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Cos
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Montesinos
J L
JL
Valero
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F
D019149:Bioreactors; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis / Q000379:methods*; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D008049:Lipase / Q000096:biosynthesis* / Q000235:genetics; D000432:Methanol / Q000032:analysis* / Q000378:metabolism*; D009862:Online Systems / Q000295:instrumentation*; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015202:Protein Engineering / Q000295:instrumentation; D011994:Recombinant Proteins / Q000096:biosynthesis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D011994:Recombinant Proteins; D008049:Lipase; D000432:Methanol