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Reduction of proteolytic breakdown in microbial homogenates
Developments in Biological Standardization
|January 1, 1985
Summary
Proteolytic enzymes in biological raw materials like E. coli and yeast show high activity. Most activity can be removed using hydrophobic gel adsorption, aiding polypeptide production.
Area of Science:
- Biochemistry
- Biotechnology
Background:
- Proteolytic enzymes are prevalent in biological raw materials essential for protein and peptide production.
- These enzymes remain active in various sources, including bacterial and yeast homogenates, ascites fluid, and cell culture media for monoclonal production.
Purpose of the Study:
- To quantify protease activity in E. coli and baker's yeast homogenates using a novel protease substrate.
- To investigate the impact of cooling and protease inhibitors on proteolytic activity.
- To assess the effectiveness of hydrophobic gel adsorption for removing protease activity.
Main Methods:
- Utilized a new protease substrate to measure protease activity.
- Evaluated the effect of temperature reduction and protease inhibitors.
- Employed hydrophobic gel media for protease adsorption.
Main Results:
- Protease activity in E. coli and baker's yeast homogenates was determined.
- Cooling and protease inhibitors had minimal impact on proteolytic activity.
- A significant portion of proteolytic activity was adsorbed onto hydrophobic gel media.
- Protease adsorption was characterized across varying temperatures and ionic strengths.
Conclusions:
- Proteolytic enzymes in E. coli and yeast present challenges for polypeptide production.
- Hydrophobic gel adsorption is an effective method for reducing protease activity.
- Understanding protease behavior is crucial for optimizing biopharmaceutical production processes.