Did you know ? If you order before Friday 14h we deliver 90PCT of the the time next Tuesday, GENTAUR another in time delivery

Pubmed ID :29132905
Publication Date : //

Purification and biochemical characterization of a 22-kDa stable cysteine- like protease from the excretory-secretory product of the liver fluke Fasciola hepatica by using conventional techniques.


This study describes the purification and characterization of a stable protease activity isolated from Fasciola hepatica adult worms maintained in vitro by employing acetone precipitation (40-60%) followed by a gel filtration through Sephadex G-100 and DEAE- cellulose ion exchange column. Through this three-step purification, the enzyme was purified 11-fold with a specific activity of 1893.9U/mg and 31.5% recovery. After the final ultrafiltration step, the purification fold was increased up to 13.1 and the overall activity yield reached a rate of 18.8%. The MW of the purified protease was estimated by reducing SDS-PAGE to be 22kDa while the proteolytic activity detection was carried out by zymography on non-denaturing SDS-PAGE containing the casein as substrate. Using this substrate, the protease showed extreme proteolytic activity at pH 5.5 and temperature 35-40°C and was highly stable over a wide range of pH, from 5.0 to 10.0. In addition to its preference for the Z-Phe-Arg-AMC fluorogenic substrate resulting in maximum proteolytic activity (99.7%) at pH 7.0, the pure protease exhibited highest cleavage activity against hemoglobin and casein substrates at pH 5.5 (85.6% and 82.8%, respectively). The Km values obtained for this protease were 5.4, 13, 160 and approximately 1000μM using respectively the fluorogenic substrate Z-Phe-Arg-AMC, hemoglobin, casein and albumin. The protease activity was completely inhibited either by E-64 inhibitor (5mM) or iodoacetamide (10mM), indicating its cysteine nature. The usefulness of the purified protease as an antigen was studied by immunoblotting. Thus, sera from sheep experimentally infected with F. hepatica recognized the protease band at 2 weeks post-infection (WPI) and strongly at 7 WPI. The early detection of antibodies anti- F. hepatica suggests the application of this molecule as a specific epitope for the serodiagnosis of fascioliasis disease.

Authors : Hemici Ahmed , Benerbaiha Roumaila Sabrina , Bendjeddou Dalila ,

Related products :

Catalog number Product name Quantity
PIS24 FASCIOLA Hepatica IgG CE 96
DEVB086 Fasciola hepatica Ag bovineELISA 96
DEVB015 Fasciola hepatica Ab bovineELISA 96
DEVB086 Fasciola hepatica Ag bovine ELISA 96 tests
DEVB015 Fasciola hepatica Ab bovine ELISA 96 tests
1460 FASCIOLA HEPATICA ANTIBODIES (DISTOMATOSIS), SERUM 1
ELI-6368 Fasciola hepatica (human) IgG ELISA with CE mark; incub. time 60_30_10 min;Sample Serum 96 Wells
EIAAB09410 CocoaCrisp,CRISP10,CRISP-10,CRISPLD1,Cysteine-rich secretory protein 10,Cysteine-rich secretory protein LCCL domain-containing 1,Homo sapiens,Human,LCCL domain-containing cysteine-rich secretory prote
20-321-175120 SECRETORY LEUCOCYTE PROTEASE INHIBITOR (SLPI) - MONOCLONAL ANTIBODY TO HUMAN SECRETORY LEUCOCYTE PROTEASE INHIBITOR (SLPI); ALP; Secretory leukocyte protease inhibitor; HUSI-1; Seminal proteinase inhi 0.1 mg
18-321-177029 SECRETORY LEUCOCYTE PROTEASE INHIBITOR (SLPI) - POLYCLONAL ANTIBODY TO HUMAN SECRETORY LEUCOCYTE PROTEASE INHIBITOR (SLPI); ALP; Secretory leukocyte protease inhibitor; HUSI-1; Seminal proteinase inhi 0.1 mg
20-321-175121 SECRETORY LEUCOCYTE PROTEASE INHIBITOR (SLPI) - BIOTINYLATED MONOCLONAL ANTIBODY TO HUMAN SECRETORY LEUCOCYTE PROTEASE INHIBITOR (SLPI); ALP; Secretory leukocyte protease inhibitor; HUSI-1; Seminal pr 0.05 mg
EIAAB09414 CRISP11,CRISP-11,CRISPLD2,Cysteine-rich secretory protein 11,Cysteine-rich secretory protein LCCL domain-containing 2,Homo sapiens,Human,LCCL domain-containing cysteine-rich secretory protein 2,LCRISP
310101 Liver Fluke IgM ELISA Kit 96T
310100 Liver Fluke IgG ELISA Kit 48T
310100 Liver Fluke IgG ELISA Kit 96T
310101 Liver Fluke IgM ELISA Kit 48T
orb90370 Intestinal Parasite Panel B Intestinal Parasite Panel B (Clonorchis sinensis, Fasciola hepatica, Schistosoma hematobium, Schistosoma mansoni) is a proficiency and educational panels for infectious dis 5x0.75 ml
EIAAB42593 Adrenal secretory serine protease,Airway trypsin-like protease,AsP,AT,Rat,Rat,Rattus norvegicus,Tmprss11d,Transmembrane protease serine 11D
EIAAB42592 Adrenal secretory serine protease,Airway trypsin-like protease,AsP,AT,Mat,Mouse,Mus musculus,Tmprss11d,Transmembrane protease serine 11D
E2015h ELISA Apoptotic cysteine protease,Apoptotic protease Mch-5,CAP4,CASP8,CASP-8,Caspase-8,FADD-homologous ICE_ced-3-like protease,FADD-like ICE,FLICE,Homo sapiens,Human,ICE-like apoptotic protease 5,MACH 96T
U2015h CLIA Apoptotic cysteine protease,Apoptotic protease Mch-5,CAP4,CASP8,CASP-8,Caspase-8,FADD-homologous ICE_ced-3-like protease,FADD-like ICE,FLICE,Homo sapiens,Human,ICE-like apoptotic protease 5,MACH, 96T
U2015h CLIA kit Apoptotic cysteine protease,Apoptotic protease Mch-5,CAP4,CASP8,CASP-8,Caspase-8,FADD-homologous ICE_ced-3-like protease,FADD-like ICE,FLICE,Homo sapiens,Human,ICE-like apoptotic protease 5, 96T
EIAAB37886 DEN1,Deneddylase-1,FKSG8,Homo sapiens,Human,NEDD8-specific protease 1,NEDP1,Protease, cysteine 2,PRSC2,SENP8,Sentrin_SUMO-specific protease SENP8,Sentrin-specific protease 8
E2015h ELISA kit Apoptotic cysteine protease,Apoptotic protease Mch-5,CAP4,CASP8,CASP-8,Caspase-8,FADD-homologous ICE_ced-3-like protease,FADD-like ICE,FLICE,Homo sapiens,Human,ICE-like apoptotic protease 5 96T
orb21888 Goat Serum Against Cat Secretory IgA Polyclonal Precipitating polyclonal goat antiserum to cat secretory IgA. In precipitating techniques as immunoelectrophoresis and radial immunodiffusion to identif 1 ml


 

GENTAUR Belgium BVBA BE0473327336
Voortstraat 49, 1910 Kampenhout BELGIUM
Tel 0032 16 58 90 45

Fax 0032 16 50 90 45
info@gentaur.com | Gentaur





GENTAUR Ltd.
Howard Frank Turnberry House
1404-1410 High Road
Whetstone London N20 9BH
Tel 020 3393 8531 Fax 020 8445 9411
uk@gentaur.com | Gentaur

 

 




GENTAUR France SARL
9, rue Lagrange, 75005 Paris
Tel 01 43 25 01 50

Fax 01 43 25 01 60
RCS Paris B 484 237 888

SIRET 48423788800017

BNP PARIBAS PARIS PL MAUBERT BIC BNPAFRPPPRG

france@gentaur.com | Gentaur

GENTAUR GmbH
Marienbongard 20
52062 Aachen Deutschland
Support Karolina Elandt
Tel: 0035929830070
Fax: (+49) 241 56 00 47 88

Logistic :0241 40 08 90 86
Bankleitzahl 39050000
IBAN lautet DE8839050000107569353
Handelsregister Aachen HR B 16058
Umsatzsteuer-Identifikationsnummer *** DE 815175831
Steuernummer 201/5961/3925
de@gentaur.com | Gentaur

GENTAUR U.S.A
Genprice Inc, Logistics
547, Yurok Circle
San Jose, CA 95123
CA 95123
Tel (408) 780-0908,
Fax (408) 780-0908,
sales@genprice.com

Genprice Inc, Invoices and accounting
6017 Snell Ave, Ste 357
San Jose, CA 95123




GENTAUR Nederland BV
NL850396268B01 KVK nummer 52327027
Kuiper 1
5521 DG Eersel Nederland
Tel:  0208-080893  Fax: 0497-517897
nl@gentaur.com | Gentaur
IBAN: NL04 RABO 0156 9854 62   SWIFT RABONL2U






GENTAUR Spain
tel:0911876558
spain@gentaur.com | Gentaur






ГЕНТАУЪР БЪЛГАРИЯ
ID # 201 358 931 /BULSTAT
София 1000, ул. "Граф Игнатиев" 53 вх. В, ет. 2
Tel 0035924682280 Fax 0035924808322
e-mail: Sofia@gentaur.com | Gentaur
IBAN: BG11FINV91501014771636
BIC: FINVBGSF

GENTAUR Poland Sp. z o.o.


ul. Grunwaldzka 88/A m.2
81-771 Sopot, Poland
TEL Gdansk 058 710 33 44 FAX  058 710 33 48              

poland@gentaur.com | Gentaur

Other countries

Österreich +43720880899

Canada Montreal +15149077481

Ceská republika Praha +420246019719

Danmark +4569918806

Finland Helsset +358942419041

Magyarország Budapest +3619980547

Ireland Dublin+35316526556

Luxembourg+35220880274

Norge Oslo+4721031366

Sverige Stockholm+46852503438

Schweiz Züri+41435006251

US New York+17185132983

GENTAUR Italy
SRL IVA IT03841300167
Piazza Giacomo Matteotti, 6
24122 Bergamo Tel 02 36 00 65 93
Fax 02 36 00 65 94
italia@gentaur.com | Gentaur