Investigations of heat treatment to improve the isolation of intracellular enzymes

✍ Scribed by J. Schnell; M. R. Kula

• Book ID: 104744449
• Publisher: Springer
• Year: 1989
• Tongue: English
• Weight: 836 KB
• Volume: 4
• Category: Article
• ISSN: 1615-7605
• DOI: 10.1007/bf00369762

✦ Synopsis

Heating, an old fashioned method for enzyme purification, was re-investigated as a continuous process in a jacketed tube heat exchanger. Biological, physical and process-engineering parameters were studied with regard to heating during the isolation of alcoholdehydrogenase from cell homogenates of baker's yeast. The heat treatment served to coagulate cell debris and protein, thereby increasing not only the specific activity of the target enzyme in solution, but at the same time the particle size of the solids. The most important parameters of the process are the selected feed temperature, pH-value, volumetric flow rate and cell concentration. At optimal conditions a clear supernatant after centrifugation, (RCF7000), is achieved containing only 50-40% of the soluble protein and 85-75 % enzyme activity, resulting in a two-fold enrichment compared to the unheated crude extract. The experiments demonstrate that a continuous heating process can be applied for pre-purification and conditioning. Enzymes with better heat stability will give even higher yields.