Abstract
An automated reverse flow injection analysis (r-FIA) system using stop-flow technique for quantifying methanol based on the enzymatic reactions of alcohol oxidase and peroxidase was developed. The system permitted methanol analysis in a linear range of 0.006-0.1 g methanol l−1 without external dilution, and with a sampling frequency of 12 analyses per hour, with a relative standard deviation of 1.16%. The analyser was validated analysing samples from a Pichia pastoris fermentation producing a heterologous protein.
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References
Alonso J, Bars J, Bartrolí J, Sánchez J, del Valle M (1995) Ionselective electrodes responsive to anionic surfactants for flow injection analysis. Anal. Chim. Acta 308: 115-121.
García de María C, Manzano T, Duarte R, Alonso A (1995) Selective flow-injection determination of methanol using immobilized enzyme reactors. Anal. Chim. Acta 309: 241-250.
Guarna MM, Lesnicki GJ, Tam BM, Robinson J, Radziminski CZ, Hasenwinkle D, Boraston A, Jervis E, MacGilivray RTA, Turner RFB, Kilburn DG (1997) On-line monitoring and control of methanol concentration in shake-flask cultures of Pichia pastoris. Biotechnol. Bioeng. 56: 279-286.
Lin Cereghino J, Cregg JM (2000) Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol. Rev. 24: 45-66.
Minning S, Serrano A, Ferrer P, Solà C, Schmid RD, Valero F (2001) Optimization of the high-level production of Rhizopus oryzae lipase in Pichia pastoris. J. Biotechnol. 86: 59-70.
Ruzicka J, Hansen EH (1988) Flow Injection Analysis, 2nd edn. New York: Wiley.
Salgado AM, Folly ROM, Valdman B, Valero F (2000) Colorimetric method for the determination of ethanol by flow injection analysis. Biotechnol. Lett. 22: 327-330.
Sekine Y, Suzuki M, Takeuchi T, Tamiya E, Karube I (1993) Selective flow-injection determination of methanol in the presence of ethanol based on a multi-enzyme system with chemiluminiscence determination. Anal. Chim. Acta 280: 179-184.
Sreekrishna K, Brankamp RG, Kropp KE, Blankenship DT, Tsay JT, Smith PL, Wierschke JD, Subramaniam A, Birkenberger LA (1997) Strategies for optimal synthesis and secretion of heterologous proteins in the methylotrophic yeast Pichia pastoris. Gene 190: 55-62.
Van der Pool JJ, de Gooijer CD, Biselli M, Wandrey C, Tramper J (1996) Automation of selective assays for on-line bioprocess monitoring by flow-injection analysis. Trends Biotechnol. 14: 471-477.
Zhang W, Bevins MA, Plantz BA, Smith LA, Meagher MM (2000) Modeling Pichia pastoris growth onmethanol and optimising the production of a recombinant protein, the heavy-chain fragment C of botulinum neurotoxin, Serotype A. Biotechnol. Bioeng. 70: 1-8.
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Almuzara, C., Cos, O., Baeza, M. et al. Methanol determination in Pichia pastoris cultures by flow injection analysis . Biotechnology Letters 24, 413–417 (2002). https://doi.org/10.1023/A:1014554324117
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DOI: https://doi.org/10.1023/A:1014554324117