Method for isolation of aminoacylase

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

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435180, 435814, 435815, 435816, C12N 980, C12N 1108

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045322147

ABSTRACT:
Aminoacylase is isolated from mammal kidneys by comminuting and homogenizing mammal kidney in water, centrifuging to form an aqueous extract, heating the aqueous extract at 60.degree. to 80.degree. C. for 5 to 15 minutes, centrifuging, adding a salt such as ammonium sulfate to the resultant supernatant, centrifuging to separate solids, dissolving the solids in water, dialyzing and recovering active aminoacylase from the dialyzed solution. The process produces aminoacylase with high specific activity and the process requires fewer purification steps. The aminoacylase obtained may be utilized directly without any subsequent purification to produce immonobilized aminoacylase. Immobilization can be carried out by covalent bonding of the aminoacylase to a partially hydrolyzed Akrilex P type acryl amide-N,N'-methylene-bis(arylamide) copolymer. The recovered aminoacylase can be subjected to two chromatographic purification steps to produce a very pure aminoacylase.

REFERENCES:
patent: 2169749 (1956-11-01), Thompson et al.
Dixon, et al., Enzymes, Academic Press, Inc., N.Y., 1964, 2nd ed., (pp. 27-48).
Birnbaum, et al., Specificity of Amino Acid Acylases, J. Biol. Chem., vol. 194, 1952, (pp. 455-470).

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