The project title is “Studies on decolorization and detoxification of Molasses spent wash by group of lignin degrading enzymes isolated from fungus”. For this, I need to separate the above mentioned enzyme.
It depends on the source, but in principle you have to try ion-exchange chromatography or preparative electrophoresis
If your aim is to decolorise and detoxify the waste - why the need to separate? - the enzyme cocktail may work better. Further all separation techniques are expensive - ion-exchange, centrifugation based on molecular weight and aqueous two phase systems - For bioremediation cocktails of enzymes can be applied. What is your fungus and what is the dominant enzyme that it is producing? You may , however, need to concentrate the cocktail before the treatment.
I agree with Vikineswary. You can use crude enzyme preparation (culture liquid or ammonium sulphate precipitate, ultrafiltration concentrat). If you need to establish the role of individual enzymes, you can use Pycnoporus which is only laccase producer, Many fungi are only laccase and MnP producers, several (P. chrysosporium) are producers of MnP and LiP, Trametes versicolor is producer of all enzymes. Moreover, you can easily to change the ratio of individual enzymes in enzyme preparation cultivating fungi in media containing various lignocellulosic materials (see our publications).
Thanks a lot to respected all for your valuable guidance. I will keep try to do such things.
Is it possible to obtain each enzyme separately in pure form? Kindly suggest technique for the same.
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