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Technical University of Munich

One-step expression and enzyme immobilisation in cellular envelopes of Escherichia coli

Ilka Sührer, doctoral thesis Technische Universität München, 2015

Production of enzymes with combined immobilization in one process step may reduce the costs for biocatalyst preparation. Therefore, enzymes were posttranslationally anchored to the cytoplasmic membrane of E. coli. The cells were subsequently opened be expression of lysis gene E from phage PhiX174 to obtain cellular envelopes with membrane anchored enzymes in stirred-tank bioreactor processes. Thus, active biocatalysts were obtained with up to 27,000 membrane-bound enzyme molecules.

Publications

  • Sührer I, Langemann T, Lubitz W, Weuster-Botz D, Castiglione K (2015): A novel one-step expression and immobilization method for the production of biocatalytic preparations. Microbial Cell Factories 14: 180-189.
  • Sührer I, Haslbeck M, Castiglione K (2014): Asymmetric synthesis of a Fluoxetin precursor with an artificial fusion protein of a ketoreductase and a formate dehydrogenase. Proc Biochem 49: 1527-1532.
  • Hölsch K, Sührer I, Heusel M, Weuster-Botz D (2013): Engineering of formate dehydrogenase: Synergistic effect of mutations affecting cofactor specificity and chemical stability. Appl Microbiol Biotechnol 97: 2473-2481.
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