Translocation of cytoplasmic <img src='/image/spc_char/beta.gif'>-galactosidase across the inner membranes of <i style="">Kluyveromyces lactis</i>

Abstract

The translocation behaviour of cytoplasmic <img src='/image/spc_char/beta.gif'>-galactosidase to periplasmic space and through the outside cell wall across the inner membranes of <i style="">Kluveromyces lactis</i> has been investigated to optimize the cell disruption process by ultrasonication for the production and separation of intracellular target biomolecule i.e. b-galactosidase. The translocation of b-galactosidase in the cells was judged by a concept of location factor (<i style="">LF</i>), which allows the location of the enzymes to be judged within the cell and has been determined using the relative rates of the enzyme and protein during the cell disruption process. The temperature was found to be useful external stimuli for the translocation of target enzyme (<i style="">LF</i> could be increased to one or more). The <i style="">LF</i> values were maximum when cells were subjected to heat stress between 45-50°C for a specified time. The enzyme activity was also found to decrease with an increase in the temperature. Maximum enzyme activity was found to be at 45°C of the heat treatment process for translocation. The kinetics of translocation of the target enzyme across the inner membrane has been reported on the basis of the variation in the location <i style="">LF</i>.