Chapter 8. Biotechnological Processing Steps for Enzyme Manufacture

  1. Prof. Dr. Andreas Sebastian Bommarius1,
  2. Dr. Bettina R. Riebel2

Published Online: 28 JAN 2005

DOI: 10.1002/3527602364.ch8

Book Title

Biocatalysis

Biocatalysis

Additional Information

How to Cite

Bommarius, A. S. and Riebel, B. R. (2005) Biotechnological Processing Steps for Enzyme Manufacture, in Biocatalysis, Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, FRG. doi: 10.1002/3527602364.ch8

Author Information

  1. 1

    School of Chemical and Biomolecular Engineering, Parker H. Petit Biotechnology Institute, Georgia Institute of Technology, 315 Ferst Drive, N. W. Atlanta, GA 30332-0363, USA

  2. 2

    Emory University School of Medicine, Whitehead Research Building, 615 Michael Street, Atlanta, GA 30322, USA

Publication History

  1. Published Online: 28 JAN 2005
  2. Published Print: 28 JAN 2004

ISBN Information

Print ISBN: 9783527303441

Online ISBN: 9783527602360

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Keywords:

  • enzyme manufacture;
  • protein isolation;
  • protein purification;
  • fermentation;
  • fed-batch culture;
  • oxygen transport in the fermenter solution;
  • kL;
  • a;
  • kLa;
  • separation;
  • centrifugation;
  • homogenization;
  • precipitation;
  • concentration of proteins;
  • purification of proteins;
  • dialysis;
  • ultrafiltration;
  • chromatography;
  • drying;
  • spray drying;
  • lyophilization;
  • stabilization for storage;
  • biocatalyst purification;
  • alcohol dehydrogenase;
  • (R)-ADH;
  • L. brevis;
  • l-amino acid oxidase;
  • Rhodococcus opacus;
  • xylose isomerase;
  • thermoanaerobium strain

Summary

  • Introduction to Protein Isolation and Purification

  • Basics of Fermentation

    • Medium Requirements

    • Sterilization

    • Phases of a Fermentation

    • Modeling of a Fermentation

    • Growth Models

    • Fed-Batch Culture

  • Fermentation and its Main Challenge: Transfer of Oxygen

    • Determination of Required Oxygen Demand of the Cells

    • Calculation of Oxygen Transport in the Fermenter Solution

    • Determination of kL, a, and kLa

  • Downstream Processing: Crude Purification of Proteins

    • Separation (Centrifugation)

    • Homogenization

    • Precipitation

    • Aqueous Two-Phase Extraction

  • Downstream Processing: Concentration and Purification of Proteins

    • Dialysis (Ultrafiltration) (adapted in part from Blanch, 1997)

    • Chromatography

    • Drying: Spray Drying, Lyophilization, Stabilization for Storage

  • Examples of Biocatalyst Purification

    • Example 1: Alcohol Dehydrogenase [(R)-ADH from L. brevis]

    • Example 2: l-Amino Acid Oxidase from Rhodococcus opacus

    • Example 3: Xylose Isomerase from Thermoanaerobium Strain JW/SLYS

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