Large-scale purification of enzymes

MD Scawen. Large scale purification of enzymes. In Enzymes in Organic Synthesis. London Pitman Publishing, 1985, pp 40-56. 

ATP extraction has been used for the separation and large-scale purification of enzymes.45 47 Proteins and nucleic acids partition between the two phases differently due to differences in their surface charges and hydrophobic-hydrophilic domains. Proteins generally prefer the aqueous phase rich in... 

Large scale purification of enzymes in PEG/dextran and PEG/salt have been reported and reviewed by Kula et. al.(18. 19. 20). Examples include pullulanase and 1,4-j-glucan phosphorylase <21 , a-amylase (22), b-galactosidase (jg3), and the general economics of extractive enzyme recovery (24). Process studies have covered the use of continuous centrifuges (18. 19) and countercurrent distribution trains (2j 25). 

F. S. Furbish, H. R Blaii I. ShOoach, R G. Pentdiev, and R. O. Brody. Enzyme replacement therapy in Gaucher s disease large-scale purification of ghicocoe-bromdase suitable for human administration. Pmc. Nail Acad. ScL USA 74 3560... 

Typical biological compounds that are separated by HIC include, cytochrome P-450, enzymes, DNA polymerase, epidermal growth factor, glycoprotein hormones, human immunoglobulins, human recombinant DNA and canine pancreatic juice proteins. Many HIC techniques have been used for large scale purification of proteins. 

Budtz P (1994) Microbial rennets for cheese making. Novo-Nordisk Technical Report, A-6045a Burgess RR (1969) A new method for the large scale purification of Escherichia coli deoxyribonucleicacid-dependent ribonucleic acid polymerase. J Biol Chem 244(22) 6160-6167 Cao L (2005) Carrier-bound immobilized enzymes. Principles, application and design. Wiley-VCH, Weinheim, 563 pp... 

Many reports on IMAC used for purifying pharmaceutically interesting proteins, such as interferons, vaccines and antibodies, have been published but relatively few data exist on actual large-scale purifications of pharmaceutical proteins. However, IMAC offers possibilities for large-scale purification of many industrial enzymes as well as proteins for research in genetics, molecular biology and biochemistryi" 1. 

The biotransformation process has been improved by significant advances in biochemical engineering advances in genetic and protein engineering, microbiological manipulations for the production of enzymes, and the use of biocatalysts in immobilized form and large-scale purification methods. 

Melling, J. and Phillips, B.W. (1975) Large scale extraction and purification of enzymes. In Handbook of Enzyme Biotechnology, edited by A.Wieseman. Sussex Ellis Horwood Ltd. 

A large scale preparation of E. coli 045 was subjected to enzyme purification using the assay for 3,5-epimerase. Protamin sulfate precipitation, ammonium sulfate fractionation was followed by DEAE-chroma-tography. The fraction containing enzymatic activity, as measured by tritium exchange, was eluted from the DEAE column early. This fraction was incapable of producing any net synthesis of TDP-6-deoxy-L-... 

R. K. Gerding and R. G. Wolfe, J. Biol. Chem., 244 1164 (1%9). Malic Dehydrogenase. VIII. Large Scale Purification and Properties of Supernatant Pig Heart Enzyme. 

---