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Proteolytic processing of penicillin amidase from Alcaligenes faecalis cloned in E. coli yields several active forms

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Abstract

Of four enzymatically active forms of Alcaligenes faecalis penicillin amidase (EC 3.5.1.11) observed in sonicated cells, two (PA5.5 and PA5.3; subscript denotes pI) could be isolated and purified in two steps from the cells destroyed by osmotic shock. Active enzyme was only found in the periplasm. PA5.5 converts further to PA5.3 which differs in the molecular mass of the A-chain. The origin of these differences is a conversion of the N-terminal Gln to pyrolideno-carboxilic acid and a loss of three amino acids from the C-terminus of the A-chain. PA5.3 had higher specific activity (10–30%) for the hydrolysis of benzylpenicillin and 6-nitro-3-phenylacetamidobenzoic acid than PA5.5. © Rapid Science Ltd. 1998

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Authors and Affiliations

  1. Department of Biotechnology II, Technical University Hamburg-Harburg, Denickestr, Hamburg, Germany

    Z. Ignatova, B. Galunsky, C. Hörnle, E. Piotraschke & V. Kasche

  2. Department of Physical Biochemistry, Physical-Chemical Institute, University Tübingen, Hoppe-Seyler-Str, Tübingen, Germany

    S. Stoeva & W. Voelter

  3. Institute of Molecular and Cell Biology, Tartu University and Estonian Biocentre, Riia Str, Tartu, Estonia

    A. Nurk

Authors
  1. Z. Ignatova
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  2. S. Stoeva
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  3. B. Galunsky
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  4. C. Hörnle
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  5. A. Nurk
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  6. E. Piotraschke
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  7. W. Voelter
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  8. V. Kasche
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Ignatova, Z., Stoeva, S., Galunsky, B. et al. Proteolytic processing of penicillin amidase from Alcaligenes faecalis cloned in E. coli yields several active forms. Biotechnology Letters 20, 977–982 (1998). https://doi.org/10.1023/A:1005410327829

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