Cistein proteaza (kaparin) iz kapara (Capparis spinosa)

Proteases are enzymes that perform very important functions in organisms and are used for a variety of objectives in vitro. In recent years, proteases have been used for clinical, pharmaceutical (alimentary digestion, anti-inflammatory, etc.) and industrial applications (cheese production, meat tenderizing, leather tanning). In this research, a protease has been purified from capsules of caper (Capparis spinosa) and characterized. Caper plants have been used for food and medicine since ancient times. The plant grows abundantly in certain regions of Turkey. Ammonium sulphate fractionation and a CM Sephadex column were used for purification of the enzyme. The purification enzyme has an optimum pH=5.0 and its optimum temperature was 60 °C. The vmax and Km values determined by Lineweaver-Burk graphics were 1.38 μg/(L·min) and 0.88 μg/L, respectively. The purification degree and the molecular mass of the enzyme (46 kDa) were determined by SDS-PAGE and gel filtration chromatography. It was investigated whether the purified and characterized protease could cause milk to congeal or digest chicken and cow meat. The results show that protease can be used for industrial production.Proteaze su enzimi koji imaju vrlo važnu funkciju u organizmu i razne mogućnosti primjene in vitro. Posljednjih se godina sve više primjenjuju u medicini, farmaciji (u liječenju probavnih tegoba, raznih upala i dr.) i industriji (u proizvodnji sira, mekšanju mesa i štavljenju kože). U ovom je radu opisan postupak pročišćavanja i karakterizacije proteaze iz kapara (Caparis spinosa). Kapare se od davnine koriste kao hrana i u medicinske svrhe, a obilato rastu u nekim dijelovima Turske. Pročišćavanje enzima provedeno je frakcioniranjem pomoću amonijeva sulfata i kromatografijom u CM-Sephadex koloni. Optimalna pH-vrijednost pročišćenog enzima bila je 5, a optimalna temperatura 60 °C. Pomoću dijagrama Lineweaver-Burke određene su vrijednosti vmax od 1,38 μg/(L×min) i Km od 0,88 μg/L. Stupanj pročišćavanja i molekularna masa enzima od 46 kDa utvrđeni su SDS-PAGE elektroforezom i gel-filtracijskom kromatografijom. Također je istraženo može li pročišćeni i karakterizirani enzim zgrušati mlijeko ili probaviti govedinu i piletinu. Rezultati su pokazali da se proteaza može primijeniti u industrijskoj proizvodnji hrane

Cistein proteaza (kaparin) iz kapara (Capparis spinosa)

Proteases are enzymes that perform very important functions in organisms and are used for a variety of objectives in vitro. In recent years, proteases have been used for clinical, pharmaceutical (alimentary digestion, anti-inflammatory, etc.) and industrial applications (cheese production, meat tenderizing, leather tanning). In this research, a protease has been purified from capsules of caper (Capparis spinosa) and characterized. Caper plants have been used for food and medicine since ancient times. The plant grows abundantly in certain regions of Turkey. Ammonium sulphate fractionation and a CM Sephadex column were used for purification of the enzyme. The purification enzyme has an optimum pH=5.0 and its optimum temperature was 60 °C. The vmax and Km values determined by Lineweaver-Burk graphics were 1.38 μg/(L·min) and 0.88 μg/L, respectively. The purification degree and the molecular mass of the enzyme (46 kDa) were determined by SDS-PAGE and gel filtration chromatography. It was investigated whether the purified and characterized protease could cause milk to congeal or digest chicken and cow meat. The results show that protease can be used for industrial production.Proteaze su enzimi koji imaju vrlo važnu funkciju u organizmu i razne mogućnosti primjene in vitro. Posljednjih se godina sve više primjenjuju u medicini, farmaciji (u liječenju probavnih tegoba, raznih upala i dr.) i industriji (u proizvodnji sira, mekšanju mesa i štavljenju kože). U ovom je radu opisan postupak pročišćavanja i karakterizacije proteaze iz kapara (Caparis spinosa). Kapare se od davnine koriste kao hrana i u medicinske svrhe, a obilato rastu u nekim dijelovima Turske. Pročišćavanje enzima provedeno je frakcioniranjem pomoću amonijeva sulfata i kromatografijom u CM-Sephadex koloni. Optimalna pH-vrijednost pročišćenog enzima bila je 5, a optimalna temperatura 60 °C. Pomoću dijagrama Lineweaver-Burke određene su vrijednosti vmax od 1,38 μg/(L×min) i Km od 0,88 μg/L. Stupanj pročišćavanja i molekularna masa enzima od 46 kDa utvrđeni su SDS-PAGE elektroforezom i gel-filtracijskom kromatografijom. Također je istraženo može li pročišćeni i karakterizirani enzim zgrušati mlijeko ili probaviti govedinu i piletinu. Rezultati su pokazali da se proteaza može primijeniti u industrijskoj proizvodnji hrane

Does Trade Foster Employment Growth in Emerging Markets? Evidence from Turkey

This work investigates the impact of importing, exporting, and two-way trading on firm labor demand in Turkish manufacturing. Adopting Multiple Propensity Score Matching techniques and Difference in Difference estimator, we support the positive internationalization impact on firm employment for an emergent country. Our evidence reveals the existence of complementarity effects between exports and imports, which is strengthened for high trade intensity firms. Furthermore, only high intensity exporting seems to promote the workforce skill upgrading in terms of an increase in the R&D worker share. The employment creation effect of firm internationalization reflects its large positive impact on firm production scale. © 2013 Elsevier Ltd

Volume CXIV, Number 4, November 7, 1996

Objective: Turner syndrome (TS) is a chromosomal disorder caused by complete or partial X chromosome monosomy that manifests various clinical features depending on the karyotype and on the genetic background of affected girls. This study aimed to systematically investigate the key clinical features of TS in relationship to karyotype in a large pediatric Turkish patient population.Methods: Our retrospective study included 842 karyotype-proven TS patients aged 0-18 years who were evaluated in 35 different centers in Turkey in the years 2013-2014.Results: The most common karyotype was 45,X (50.7%), followed by 45,X/46,XX (10.8%), 46,X,i(Xq) (10.1%) and 45,X/46,X,i(Xq) (9.5%). Mean age at diagnosis was 10.2±4.4 years. The most common presenting complaints were short stature and delayed puberty. Among patients diagnosed before age one year, the ratio of karyotype 45,X was significantly higher than that of other karyotype groups. Cardiac defects (bicuspid aortic valve, coarctation of the aorta and aortic stenosis) were the most common congenital anomalies, occurring in 25% of the TS cases. This was followed by urinary system anomalies (horseshoe kidney, double collector duct system and renal rotation) detected in 16.3%. Hashimoto's thyroiditis was found in 11.1% of patients, gastrointestinal abnormalities in 8.9%, ear nose and throat problems in 22.6%, dermatologic problems in 21.8% and osteoporosis in 15.3%. Learning difficulties and/or psychosocial problems were encountered in 39.1%. Insulin resistance and impaired fasting glucose were detected in 3.4% and 2.2%, respectively. Dyslipidemia prevalence was 11.4%.Conclusion: This comprehensive study systematically evaluated the largest group of karyotype-proven TS girls to date. The karyotype distribution, congenital anomaly and comorbidity profile closely parallel that from other countries and support the need for close medical surveillance of these complex patients throughout their lifespa

Discutindo a educação ambiental no cotidiano escolar: desenvolvimento de projetos na escola formação inicial e continuada de professores

A presente pesquisa buscou discutir como a Educação Ambiental (EA) vem sendo trabalhada, no Ensino Fundamental e como os docentes desta escola compreendem e vem inserindo a EA no cotidiano escolar., em uma escola estadual do município de Tangará da Serra/MT, Brasil. Para tanto, realizou-se entrevistas com os professores que fazem parte de um projeto interdisciplinar de EA na escola pesquisada. Verificou-se que o projeto da escola não vem conseguindo alcançar os objetivos propostos por: desconhecimento do mesmo, pelos professores; formação deficiente dos professores, não entendimento da EA como processo de ensino-aprendizagem, falta de recursos didáticos, planejamento inadequado das atividades. A partir dessa constatação, procurou-se debater a impossibilidade de tratar do tema fora do trabalho interdisciplinar, bem como, e principalmente, a importância de um estudo mais aprofundado de EA, vinculando teoria e prática, tanto na formação docente, como em projetos escolares, a fim de fugir do tradicional vínculo “EA e ecologia, lixo e horta”.Facultad de Humanidades y Ciencias de la Educació

CARBONIC ANHYDRASE FROM BOVINE BONE

In this research, carbonic anhydrase enzyme, which was taken from the bones of an animal, was purified and characterized for the first time. For this, the bones of a young cow were used. The purification treatment was completed in three steps. Three different isoenzymes, such as peripheral, cystolic, and integral from the bone-cell cytozolic isoenzyme were purified and characterized. In purification of the three isoenzymes, the technique of affinity chromatography, which utilized Sepharose-4B-L-Tyrosine-Sulphanylamide, was used. In measuring the activities of enzymes, two different methods were applied. These are the esterase methods that utilize hydratase and p-nitrophenylacetate as substrate. The measurement of proteins was done with the methods of Bradford and Coomassie Brillant Blue. The optimum pH and temperature of each enzyme were measured and molecular weights were measured by gel-filtration. Its purity was examined by SDS-PAGE (3-10% alternating) electrophoresis and the inferior unit was defined. The inhibition effects of some chemicals were tested for each of the three isoenzymes

Carbonic Anhydrase from Potato (Solanum tuberosum) Roots and Leaves

Carbonic anhydrases (CA: carbonate hydrolase: E.C.4.2.1.1) from leaves and roots of mature potato (Solarium tuberosum) were purified and characterized. The purification levels of enzymes were 87.91 fold and 40.601 fold in leaves and roots, respectively. The optimum temperature was 40 degrees C in leaves and in roots. pHs optimal were 8.5 and 11 in leaves and in roots, respectively. Enzymes of leaves were formed 5 monomers that it's having molecular weights of 22,000, 28,000, 35,000, 40,000, 65,000, 5 polymers that it's having 73,000, 80,000, 83,000, 132,000 and 200,000 Dalton and these proteins had carbonic anhydrase activity. But there is at the levels of 22,000, 28,000, 35,000 and 132,000 Dalton in gel filtration, for leaves. SDS-PAGE was done for roots and leaves and subunits were obtained. In addition the enzyme against the effect of NaNa(3), KSCN and sulphanylamide, which was known as an inhibitor of mammalian carbonic anhydrase, was determined. Potato (Solanum tuberosum) has ascorbic acid (vitamin C) in very high level and iron and calcium ions in low level. So, the effect of ascorbic acid, FeCl(3) and CaCl(2) in different concentrations on enzyme was determined for roots and leaves, separately. Finally, carbonic anhydrase was purified from roots and leaves of potato (Solanum tuberosum), separately and they were done optimal. Carbonic anhydrase functions in respiration and has a part in photosynthesis in plants. It was a serious deficiency that carbonic anhydrase wasn't defined in potato (Solanum tuberosum)

Purification and some properties of carbonic anhydrase from Elephas trogontherii (Steppe elephant) bone

Four isoenzymes of carbonic anhydrase (CA) were purified from Elephas trogontherii (steppe elephant) bone (approx 0.3-0.5 million years old) from different locations (outer peripheral, cytosolic, inner peripheral and integral) using Sepharose 4B-L-tyrosine sulphanilamide affinity chromatography and their kinetics properties were investigated and compared with known CA isoenzymes. The purification degree of CAs was monitored by SDS-PAGE. Purification fold for outer peripheral, inner peripheral, cytosolic and integral CA was 395.6, 652.8, 1091 and 429.3 and the molecular mass (as determined by gel filtration chromatography) was 37, 36, 35, and 39 kDa, respectively. The optimal temperature for isozymes was 10-20, 30, 30 and 60 degrees C and optimal pH was between 7.5-11, 7.5-10, 7.5-10 and 7.5 respectively. K values (at optimum pH and 20 degrees C) for p-nitrophenyl acetate as substrate were 4.83, 6.80, 4.525 and 3.86 mM and the V,,,,, values for the same substrate were 0.00097, 0.0149, 0.00249 and 0.00072 mu mol/L*mm, respectively. I-50 values of isoenzymes for the inhibitors of CA - sulphanilamide, KSCN, acetazolamide and NaN3 were also determined

Purification of Paraoxonase (PON1) from Olive (Olea europaea L.) and Effect of Some Chemicals on Paraoxonase Activity in vitro

Paraoxonase was purified from olive (Olea europaea L.) using sepharose 4B-L-tyrosine-1-naphthylamine affinity chromatography. This enzyme was purified 173.4-fold. SDS-polyacrylamide electrophoresis of the enzyme indicates a single protein staining band with an apparent Mr of 45 kDa. The kinetic properties of the purified enzyme were determined. The enzyme exhibited high activity at broad pH (pH 5.0-9.0) and temperature (40 and 70 degrees C). The purified enzyme remained stable at 4 degrees C for more than I year. Paraoxonase was mostly stable at 40 degrees C. Its' activity decreased in 55 % for 1 h at 60 degrees C and 20 % for 4 h at 50 degrees C. Using paraoxon as a substrate, the K-m and V-max values for the purified enzyme were estimated to be 3.76 mM and 131.5 mu mol L dak(-1), respectively. The activities were strongly inhibited by Hg2+ and Fe3+, while Cu2+, beta-mercaptoethanol, dithioerythritol and SDS slightly activated the enzyme. As judged by catalytic efficiencies, paraoxon is the preferred substrate