integrated one-pot enrichment and immobilization of styrene monooxygenase (stya) using sepabead ec-ea and ec-q1a anion-exchange carriers苯乙烯综合锅浓缩和固定的单氧酶(stya)使用sepabead ec-ea和ec-q1a阴离子交换运营商.pdfVIP

Molecules 2011, 16, 5975-5988; doi:10.3390/molecule OPEN ACCESS molecules ISSN 1420-3049 /journal/molecules Article Integrated One-Pot Enrichment and Immobilization of Styrene Monooxygenase (StyA) Using SEPABEAD EC-EA and EC-Q1A Anion-Exchange Carriers Reto Ruinatscha, Rohan Karande, Katja Buehler and Andreas Schmid * Department of Chemical and Biochemical Engineering, Chair of Chemical Biotechnology, TU Dortmund, Dortmund 44221, Germany * Author to whom correspondence should be addressed; E-Mail: andreas.schmid@bci.tu-dortmund.de; Tel.: +49-231-755-7380; Fax: +49-231-755-7382. Received: 23 June 2011 / Accepted: 12 July 2011 / Published: 18 July 2011 Abstract: A straightforward one-pot procedure combining enrichment and immobilization of recombinantely expressed FADH2 dependent styrene monooxygenase (StyA) directly from Escherichia coli cell extracts was investigated. Sepabeads EC-EA and EC-Q1A anion-exchange carriers were employed to non-covalently adsorb StyA from the cell extracts depending on basic parameters such as varying initial protein concentrations and pH. The protein fraction of the cell extract contained around 25% StyA. At low initial protein concentrations (2.5 mg mL−1) and pH 6, the enzyme could be enriched up to 52.4% on Sepabeads EC-EA and up to 46.0% on Sepabeads EC-Q1A, accounting for an almost complete StyA adsorption from the cell extracts. Higher initial protein concentrations were necessary to exploit the high loading capacity of