Dear Sir. Concerning your issue about the extraction of intra cellular enzymes for industrial application. Enzymes are the biocatalysts synthesized by living cells. They are complex protein molecules that bring about chemical reactions concerned with life. It is fortunate that enzymes continue to function (bring out catalysis) when they are separated from the cells i.e. in vitro. Basically, enzymes are non­toxic and biodegradable. They can be produced in large amounts by microorganisms for industrial applications. Enzyme technology broadly involves production, isolation, purification and use of enzymes (in soluble or immobilized form) for the ultimate benefit of humankind. In addition, recombinant DNA technology and protein engineering involved in the production of more efficient and useful enzymes are also a part of enzyme technology. The commercial production and use of enzymes is a major part of biotechnology industry. The specialties like microbiology; chemistry and process engineering, besides biochemistry have largely contributed for the growth of enzyme technology. I think the following below links may help you in your analysis:

http://www.biologydiscussion.com/enzymes/enzyme-technology/enzyme-technology-application-and-commercial-production-of-enzymes/10185

http://biologymad.com/StudentsWork/12%20-%20etnotes.pdf

http://www.isca.in/IJBS/Archive/v2/i10/8.ISCA-IRJBS-2013-153.pdf

https://www.eolss.net/Sample-Chapters/C17/E6-58-05-01.pdf

Thanks

Proteins are very complicated macromolecules, and while some of them are stable at a range of pH, temperatures, and pressures, many are very unstable and require optimization at each step of purification. Since you are focusing specifically on enzymes, it seems reasonable to purify your target of interest using specific activity. Specific activity guided fractionation allows you to fractionate the cellular protein and test for enzymatic activity of each the resultant fractions. I recommend starting with a gentle lysis procedure (I prefer pulse sonication on ice in lysis buffer with salt) followed by centrifugation to remove cellular debris. In this instance you have two fractions the cleared lysate and the lysate pellet. Resuspend the pelleting the same volume the cleared lysate and test for enzymatic activity. Then proceed to the next separation with the enzymatically active fraction. Truthfully, a great place to start is a literature search for your enzyme class to find some purification schemes that have worked for similar proteins. You'll want to optimize the enzyme activity assay. 

Due to the inherent complexity of proteins, their size, flexibility, and susceptibility to proteolysis, no one technique will work for all proteins. You might want to add acetonitrile to your proteinaceous fraction and test enzymatic activity to see if the protein is still active in the presence of organic solvent. If the activity is retained then proceeding with Reverse-Phase Liquid Chromatography will yielded better resolution and often less steps to purity. Otherwise, you can take advantage of surface charge with Ion-Exchange Chromatography, hydrophobicity with Hydrophobic Interaction Chromatography, size with Size-Exclusion Chromatography, and there are many adsorptive methods that may or may not work for your system (for example). Each step will require optimization and just because your protein of interest doesn't stick to a specific column doesn't mean it isn't a worthwhile step as other proteins might be removed in the process affording you more pure protein.

Furthermore, you will probably want to add protease inhibitors during lysis and possibly through the entire purification scheme. Worst case scenario you need to purify the protein at 4 degrees C.