Purification of swine serum angiotensin converting enzyme with high recovery of activity using lisinopril coupled to epoxy‐activated sepharose 6B

Quassinti, Luana; Miano, Antonino; Bramucci, Massimo; Maccari, Ennio; Amici, Domenico

doi:10.1080/15216549800201942

Cited by 8 publications

(9 citation statements)

References 3 publications

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“…Enzyme purification is briefly reported in Materials and Methods, while the summary of the purification procedure of each serum is reported in Table 1. Recovery of activity of serum ACEs was smaller than reported for swine serum ACE [8], but similar to that reported in literature [13]. The data reported in Table 1 show that foetal equine and foetal bovine sera contain about twice as much ACE activity as corresponding adult sera.…”

Section: Resultssupporting

confidence: 86%

“…Fresh bovine serum, collected at the slaughter house, was used as comparison. A method of ACE purification, developed in our laboratory for swine serum to obtain high recovery of activity in few steps [8], was applied to sera. Enzyme purification is briefly reported in Materials and Methods, while the summary of the purification procedure of each serum is reported in Table 1.…”

Section: Resultsmentioning

confidence: 99%

“…The purification was carried out as described for ACE purification from swine serum [8]. Briefly, serum was precipitated with (NH4)2SO4 (35 -65 % of saturation).…”

Section: Methodsmentioning

confidence: 99%

“…Initial rates were measured for the first 30 rain of reaction. The separation of hydrolytic products was carried out following the reverse-phase HPLC analysis [8]. Kinetic parameters were derived from Lineweaver-Burk plots, values of Kcat being based on a molecular weight for purified ACEs of 170 kDa.…”

Section: Methodsmentioning

confidence: 99%

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Presence and comparison of angiotensin converting enzyme in commercial cell culture sera

et al. 1999

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SUMMARYThis study was conducted to determine the presence of the angiotensin converting enzyme in commercial sera used in cell culture medium. The aim of the research was to bring the presence of proteinases (angiotensin converting enzyme) to cell culture users' knowledge and to give some data for solving problems about the development of peptides as useful drugs. The enzymes, purified from foetal bovine, adult bovine, foetal equine, adult equine, and human sera, showed molecular weights of about 170 kDa. Captopril and lisinopril inhibited enzyme activities at nanomolar concentrations. The enzymes were able to hydrolyze, with different efficiency, angiotensin I, bradykinin and epidermal mitosis inhibiting pentapeptide. The heat inactivation of commercial sera at 56 ~ for 30 min showed a reduction of ACE activity of about 35-80 %. Therefore, the presence of ACE activity in commercial sera can influence the activity of biological peptides tested on cell lines cultured "in vitro".

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Section: Resultssupporting

confidence: 86%

Section: Resultsmentioning

confidence: 99%

“…The purification was carried out as described for ACE purification from swine serum [8]. Briefly, serum was precipitated with (NH4)2SO4 (35 -65 % of saturation).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Presence and comparison of angiotensin converting enzyme in commercial cell culture sera

et al. 1999

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“…Enzyme was purified by one step lisinopril affinity chromatography, carried out as reported (14). Gel electrophoresis under denaturing conditions was performed by SDS-7.5% polyacrylamide gel (8.3 em x 6 em) at constant voltage (9), and was silver-stained (20).…”

Section: Methodsmentioning

confidence: 99%

La enzima convertidora de angiotensina purificada de ovario de Rana esculenta influye en la síntesis de esteroides in vitro

Miano

Quassinti

Maccari

et al. 2003

J. Physiol. Biochem.

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The aim of the present study was to purify and characterize angiotensin-converting enzyme (ACE) present in frog ovary (Rana esculenta). Detergent and trypsin-extracted enzymes were purified using a one-step process, consisting of affinity chromatography on lisinopril coupled to Sepharose 6B. The molecular mass was 150 kDa for both detergent-extracted and trypsin-extracted enzyme. The specific activity of detergent-extracted and trypsin-extracted ACE was 294 U mg(-1) and 326 U mg(-1) respectively. The optimum pH range was from 7-8.5 at 37 degrees C and the optimum temperature was 50 degrees C. Optimum chloride concentration was about 200 mM for synthetic substrate FAPGG (N-[3-(2-furyl)acryloyl] L-phenylalanyl glycyl glycine) and angiotensin I, and 10 mM for bradykinin. The Km and Kcat values for FAPGG were 0.608 +/- 0.07 mM and 249 sec(-1) respectively and I50 values for captopril and lisinopril, two specific ACE inhibitors, were 68 +/- 12.55 nM and 6.763 +/- 0.66 nM respectively. Frog ovary tissue from prereproductive period was incubated in vitro in the presence of frog ovary ACE (2.5 mU/ml), captopril (0.1 mM), and lisinopril (0.1 mM). Production of 17beta-estradiol, progesterone, and prostaglandins E2 and F2alpha was determined. The data showed a modulation of 17beta-estradiol, progesterone and prostaglandin E2 production by ovary ACE.

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Investigation of inhibition effect of butanol and water extracts of Matricaria chamomilla L. on angiotensin‐converting enzyme purified from human plasma

Baş

Türkoğlu

Göz

2021

Biotech and App Biochem

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Angiotensin-converting enzyme (ACE) liable for the regulation of blood pressure was purified from human plasma by affinity chromatography. Impact of water and butanol extracts of Matricaria chamomilla L. on purity ACE was examined. ACE was purified using the affinity chromatography method. The enzyme activity was evaluated at 345 nm by a spectrophotometer. Extracts of M. chamomilla plant with butanol and water were prepared. Lisinopril was utilized as a specific inhibitor. ACE was purified 3,659-fold from human plasma and the specific activity was 1,350 EU/mg protein. The molecular weight and purity of ACE were found by SDS-PAGE and two bands of 60 and 70 kDa on the gel were detected. Water and butanol extracts of M. chamomilla demonstrated inhibitor impact on ACE activity. IC 50 constants for water and butanol extracts of M. chamomilla were computed to be 1.292 and 0.353 mg/mL, respectively. The type of inhibition for whole inhibitors was identified as noncompetitive. IC 50 and K i constants for lisinopril were calculated to be 0.781 and 0.662 nM, respectively. These results indicate that butanol and water extracts of M. chamomilla may have an ACE inhibitor potential.

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Purification of swine serum angiotensin converting enzyme with high recovery of activity using lisinopril coupled to epoxy‐activated sepharose 6B

Cited by 8 publications

References 3 publications

Presence and comparison of angiotensin converting enzyme in commercial cell culture sera

Presence and comparison of angiotensin converting enzyme in commercial cell culture sera

La enzima convertidora de angiotensina purificada de ovario de Rana esculenta influye en la síntesis de esteroides in vitro

Investigation of inhibition effect of butanol and water extracts of Matricaria chamomilla L. on angiotensin‐converting enzyme purified from human plasma

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