Encapsulation of PEG-urease/PEG-AlaDH enzyme system in erythrocyte.
No intravenously injectable enzyme preparate containing urease as an alternetive to hemodialysis, hemoperfusion and CAPD systems in patients having chronic renal failure has been encountered in literature. In this study, it has been aimed to convert blood urea to alanine by using PEG-urease/PEG-AlaDH enzyme pair encapsulated within living erythrocyte. In this system, urea is decomposed into NH3 and HCO3- and the ammonia released is converted into alanine by reacting pyruvate under the catalytic action of alaninedehydrogenase. The production of pyruvate and NADH by erythrocyte required in the second stage of the reaction will make the process a feasible and ceaseless one. The success of the system will enable the renal patients with diabetes mellitus. Urease and AlaDH were covalently immobilized on activated PEG. PEG-urease/PEG-AlaDH were encapsulated in erythrocyte (1/1)(v/v) by using slow dialysis methods. The activity of enzyme system, encapsulation yield and hemogram analysis were determined for each sample.
Hamarat Baysal
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Ege University, Faculty of Science, Biochemistry Department, Bornova, Izmir, Turkey. hamarat@fenfak.ege.edu.tr
Uslan
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AH
D000409:Alanine / Q000096:biosynthesis; D050788:Alanine Dehydrogenase; D000594:Amino Acid Oxidoreductases / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*; D000641:Ammonia / Q000032:analysis; D003920:Diabetes Mellitus / Q000628:therapy; D004339:Drug Compounding; D016503:Drug Delivery Systems / Q000379:methods* / Q000592:standards; D004800:Enzymes, Immobilized / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics; D004912:Erythrocytes / Q000201:enzymology*; D006403:Hematologic Tests; D006801:Humans; D007700:Kinetics; D011092:Polyethylene Glycols; D019289:Pyruvic Acid / Q000032:analysis; D014510:Urease / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics*
D004800:Enzymes, Immobilized; D011092:Polyethylene Glycols; D000641:Ammonia; D019289:Pyruvic Acid; D000594:Amino Acid Oxidoreductases; D050788:Alanine Dehydrogenase; D014510:Urease; D000409:Alanine