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Preparation of Semisynthetic Enzymes by Chemical Means

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Biotechnology and Polymers

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Abstract

A method to generate catalytic activity by conformational modification of proteins is discussed and several examples elaborated. Semisynthetic amino acid esterases and glucose isomerases are discussed briefly. The most recent studies, which are described in more detail, concern semisynthetic fluorohydrolases prepared by conformational modification of bovine pancreatic ribonuclease (RNase) as well as other proteins. RNase, modified with hexamethylphosphoramide (HMPA), was derivatized with diimidates of chain lengths from one to eight carbon atoms to determine which chain length produced the maximum fluorohydrolase activity. The highest activity is observed when RNase is crosslinked with dimethyl pimelimidate. This derivative operates over a pH range of 6.5 to 8.0 with an optimum pH of approximately 7.5 and hydrolyzes phenylmethylsulfonylfluoride (PMSF) as well as the potent acetylcholinesterase inhibitor, diisopropylfluorophosphate (DFP). The mean fluorohydrolase activity, after chromatography on G-15 Sephadex to remove reactants, was 0.8 ± 0.2 U/mg.

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Authors and Affiliations

  1. Anatrace, Inc., 1280 Dussel Drive, Maumee, OH, 43537, USA

    David E. Albert, Mary B. Douglas, Marcia A. Hintz, Christopher S. Youngen & Dr. Melvin H. Keyes

  2. Department of Biochemistry, Medical College of Ohio, P. O. Box 10008, Toledo, OH, 43699-0008, USA

    David E. Albert, Mary B. Douglas, Marcia A. Hintz, Christopher S. Youngen & Dr. Melvin H. Keyes

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  1. David E. Albert
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  2. Mary B. Douglas
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  3. Marcia A. Hintz
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  4. Christopher S. Youngen
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  5. Dr. Melvin H. Keyes
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Editors and Affiliations

  1. Department of Chemistry, Youngstown State University, Youngstown, OH, 44555, USA

    Charles G. Gebelein

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Albert, D.E., Douglas, M.B., Hintz, M.A., Youngen, C.S., Keyes, M.H. (1991). Preparation of Semisynthetic Enzymes by Chemical Means. In: Gebelein, C.G. (eds) Biotechnology and Polymers. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-3844-8_25

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  • DOI: https://doi.org/10.1007/978-1-4615-3844-8_25

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-6715-4

  • Online ISBN: 978-1-4615-3844-8

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