Gentle lysis of mucous producing cold-adapted bacteria by surfactant treatment combined with mechanical disruption.
A procedure for the enhanced lysis of mucous producing psychrotrophic gram positive bacteria for subsequent enzyme studies is described. An initial washing of bacterial cells with Tween 80 was found to improve the degree of cell disruption in subsequent sonication or grinding with glass beads, resulting in about 20-200% increase in total soluble protein content. However, in terms of lactate dehydrogenase (LDH) activity present in the lysate, pretreatment with Tween 80 was more effective in combination with grinding, especially in the highly mucous producing strain GY11. The type of surfactant used in the pretreatment procedure before grinding strongly influenced the percentage lysis of tested strains, both in terms of released soluble protein and enzyme activity. Zymograms of LDH and glutamate dehydrogenase (GDH) activity present in the lysates also very well supported the results obtained by total protein and enzyme activity measurements.
Nandakumar
R
R
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Sweden.
Gounot
A M
AM
Mattiasson
B
B
D000222:Adaptation, Physiological; D016667:Bacterial Capsules / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry; D001431:Bacteriological Techniques; D001433:Bacteriolysis; D003080:Cold Temperature; D006094:Gram-Positive Bacteria / Q000737:chemistry / Q000502:physiology*; D013010:Sonication; D013501:Surface-Active Agents / Q000494:pharmacology*
D001426:Bacterial Proteins; D013501:Surface-Active Agents