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An optimized ultrasonication protocol for bacterial cell disruption and recovery of β-galactosidase fusion proteins

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Biotechnology Techniques

Summary

In this work, we have developed and optimized an ultrasonication protocol for Escherichia coli recombinant cells, adapted to laboratory-scale release of β-galactosidase fusion proteins. After a single sonication treatment of 15 minutes, about 30% of recombinant protein present in the sample remains still associated to cellular debris, and it can not be removed by increasing the sonication time. After a clarification step a second sonication treatment of the resuspended cell debris again releases only a 70% of the remaining product. Therefore, the application of two short, consecutive sonication treatments permits a global recovery yield of about 90%. The use of a new disruption buffer to stabilize β-galactosidase allows the fusion proteins to maintain the active form throughout the process.

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Authors and Affiliations

  1. Institut de Biologia Fonamental and Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193, Barcelona, Spain

    Jordi X. Feliu & Antonio Villaverde

Authors
  1. Jordi X. Feliu
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  2. Antonio Villaverde
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Feliu, J.X., Villaverde, A. An optimized ultrasonication protocol for bacterial cell disruption and recovery of β-galactosidase fusion proteins. Biotechnol Tech 8, 509–514 (1994). https://doi.org/10.1007/BF00222845

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  • DOI: https://doi.org/10.1007/BF00222845

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