Related Experiment Video
Updated: Aug 14, 2026
13:30
A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
Published on: November 7, 2012
Factors determining more efficient large-scale release of a periplasmic enzyme from E. coli using lysozyme
J J Pierce1, C Turner, E Keshavarz-Moore
1Department of Chemical and Biochemical Engineering, University College London, UK.
Journal of Biotechnology
|October 23, 1997
Summary
This study optimized lysozyme use for releasing periplasmic enzymes from E. coli, reducing enzyme concentration and sucrose levels. This improves downstream processing and enzyme recovery, even without lysozyme.
Area of Science:
- Biotechnology
- Enzyme Technology
- Microbial Processing
Background:
- Lysozyme use for periplasmic enzyme release is limited by cost and contamination.
- Efficient recovery of periplasmic enzymes from E. coli is crucial for industrial applications.
Purpose of the Study:
- To optimize lysozyme-mediated periplasmic enzyme release from E. coli at pilot scale.
- To reduce lysozyme concentration and improve downstream processing efficiency.
Main Methods:
- Investigated effects of sucrose, lysozyme, and cell concentrations on enzyme release.
- Optimized releasing mixture components for improved yield.
- Assessed lysozyme binding to cellular debris.
Main Results:
- Reduced lysozyme concentration by 5-fold and sucrose concentration to 15% (w/v).
- Achieved high enzyme release levels with improved centrifugal harvesting.
- Demonstrated enzyme release is possible without lysozyme and significant lysozyme remains bound to cellular debris.
Conclusions:
- Optimized conditions enable cost-effective periplasmic enzyme recovery.
- Reduced lysozyme usage minimizes contamination in downstream processing.
- Lysozyme's binding to cellular debris facilitates its removal.
