[Immobilization and properties of angiotensin converting enzyme]. 2000

H Liu, and L Chen
Department of Biochemistry, Institute of Cardiovascular Disease, CAMS and PUMC, Beijing 100037, China.

OBJECTIVE To study the immobilization of angiotensin converting enzyme (ACE) for purifying ACE inhibitor from a native peptide mixture. METHODS The experiment was carried out under the low water activity condition, using tosylate chloride activating side-chain hydroxyl group of Sepharose CL-4B agarose to form a high active group which could react with the free amino-group of ACE to link the enzyme with agarose. RESULTS Immobilized ACE not only had a wider pH range, but also had a 0.6 unit right-move optimum pH than soluble ACE. After treated in pH9.0 and pH 6.5 conditions for 24 h respectively, 82% and 68% enzyme activities of immobilized ACE was maintained, and soluble ACE remained 64% and 39%. Immobilized and soluble ACE both appeared maximum enzyme activity at about 50 degrees C, the soluble ACE would lose almost all its activity when temperature kept rising. When kept at 40 degrees C and 50 degrees C for 2 h, the activity of immobilized ACE remained 82% and 34% respectively, while the soluble ACE remained 52% and completely inactivated. After two kinds of enzymes were stored at 20 degrees C for one month, immobilized ACE remained 61% activity, as compared with the 20% activity residual of soluble ACE. CONCLUSIONS The immobilized ACE had a better stability than soluble ACE in conditioned pH and temperature.

UI MeSH Term Description Entries
D007703 Peptidyl-Dipeptidase A A peptidyl-dipeptidase that catalyzes the release of a C-terminal dipeptide, oligopeptide-|-Xaa-Yaa, when Xaa is not Pro, and Yaa is neither Asp nor Glu. Thus, conversion of ANGIOTENSIN I to ANGIOTENSIN II, with increase in vasoconstrictor activity, but no action on angiotensin II. It is also able to inactivate BRADYKININ, a potent vasodilator; and has a glycosidase activity which releases GPI-anchored proteins from the membrane by cleaving the mannose linkage in the GPI moiety. (From https://www.uniprot.org April 15, 2020). ACE1 Angiotensin-Converting Enzyme 1,ACE1 Protein,Angiotensin Converting Enzyme,Angiotensin Converting Enzyme 1,Antigens, CD143,CD143 Antigens,Dipeptidyl Carboxypeptidase I,Kininase II,Peptidase P,Angiotensin I-Converting Enzyme,Carboxycathepsin,Dipeptidyl Peptidase A,Kininase A,ACE1 Angiotensin Converting Enzyme 1,Angiotensin I Converting Enzyme,Carboxypeptidase I, Dipeptidyl,Peptidyl Dipeptidase A
D004795 Enzyme Stability The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat. Enzyme Stabilities,Stabilities, Enzyme,Stability, Enzyme
D004800 Enzymes, Immobilized Enzymes which are immobilized on or in a variety of water-soluble or water-insoluble matrices with little or no loss of their catalytic activity. Since they can be reused continuously, immobilized enzymes have found wide application in the industrial, medical and research fields. Immobilized Enzymes,Enzyme, Immobilized,Immobilized Enzyme
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D013696 Temperature The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms. Temperatures

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