Purification of glucose 6-phosphate dehydrogenase from goose erytrocytes and kinetic properties
Glukoz 6-fosfat dehidrogenaz (G6PD) enzimi kaz eritrositlerinden saflaştırıldı ve bazı karakteristik özellikleri araştırıldı. Saflaştırma prosedürü;' hemolizatın hazırlanması, amonyum sülfat çöktürmesi ve 2', 5'-ADP Sepharose 4B afınite kromatografisi şeklinde üç basamaktan ibarettir....
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| Veröffentlicht in: | Turkish journal of veterinary & animal sciences 2003, Vol.27 (5), p.1179-1185 |
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| container_title | Turkish journal of veterinary & animal sciences |
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| creator | YILMAZ, Hayrullah KÜFREVİOĞLU, Ö.İrfan ÇİFTÇİ, Mehmet BAKAN, Ebubekir BEYDEMİR, Şükrü |
| description | Glukoz 6-fosfat dehidrogenaz (G6PD) enzimi kaz eritrositlerinden saflaştırıldı ve bazı karakteristik özellikleri araştırıldı. Saflaştırma prosedürü;' hemolizatın hazırlanması, amonyum sülfat çöktürmesi ve 2', 5'-ADP Sepharose 4B afınite kromatografisi şeklinde üç basamaktan ibarettir. Bu üç basamak neticesinde 36,2 EÜ/mg protein spesifik aktivitesine sahip olan enzim, % 68,79 verimle 3.892 kat saflaştırıldı ve enzim saflığının kontrol edilmesi için SDS-PAGE yapıldı. Enzim için optimum pH, stabil pH, optimum sıcaklık, molekül ağırlığı, NADP* için Km ve Vmax değerleri, G6-P için Km ve Vmax değerleri hesaplandı. Ayrıca ATP, ADP ve NADPH inhibitörleri için Lineweaver-Burk grafikleri çizilerek K, değerleri ve inhibisyon tipleri tespit edildi. Bu maddeler enzimi yarışmasız olarak inhibe etti.
Glucose 6-phosphate dehydrogenase (G6PD) was purified from goose erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of 3 steps: hemolysate preparation, ammonium sulfate precipitation, and 2', 5'-ADP Sepharose 4B affinity gel chromatography. Thanks to the 3 consecutive procedures, the enzyme, having a specific activity of 36.2 EU/mg protein, was purified for a yield of 68.79% and 3892 folds; to ascertain enzyme purity, SDS-PAGE was performed. Optimal pH, stable pH, optimal temperature, molecular weight, and Km and Vmax values for NADP* and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. In addition, K, values and inhibition type were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP and NADPH. These materials inhibited the enzyme in a noncompetitive manner. |
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Glucose 6-phosphate dehydrogenase (G6PD) was purified from goose erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of 3 steps: hemolysate preparation, ammonium sulfate precipitation, and 2', 5'-ADP Sepharose 4B affinity gel chromatography. Thanks to the 3 consecutive procedures, the enzyme, having a specific activity of 36.2 EU/mg protein, was purified for a yield of 68.79% and 3892 folds; to ascertain enzyme purity, SDS-PAGE was performed. Optimal pH, stable pH, optimal temperature, molecular weight, and Km and Vmax values for NADP* and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. In addition, K, values and inhibition type were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP and NADPH. These materials inhibited the enzyme in a noncompetitive manner.</description><identifier>ISSN: 1300-0128</identifier><language>eng</language><publisher>TÜBİTAK</publisher><subject>alyuvarlar ; Animal Physiology and Biochemistry (Excluding Nutrition) ; erythrocytes ; geese ; glucose-6-phosphate dehydrogenase ; glukoz-6-fosfat dehidrogenaz ; Hayvan Fizyolojisi ve Biyokimyası (Beslenme Dışında) ; kaz ; purification ; saflaştırma</subject><ispartof>Turkish journal of veterinary & animal sciences, 2003, Vol.27 (5), p.1179-1185</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,4024</link.rule.ids></links><search><creatorcontrib>YILMAZ, Hayrullah</creatorcontrib><creatorcontrib>KÜFREVİOĞLU, Ö.İrfan</creatorcontrib><creatorcontrib>ÇİFTÇİ, Mehmet</creatorcontrib><creatorcontrib>BAKAN, Ebubekir</creatorcontrib><creatorcontrib>BEYDEMİR, Şükrü</creatorcontrib><title>Purification of glucose 6-phosphate dehydrogenase from goose erytrocytes and kinetic properties</title><title>Turkish journal of veterinary & animal sciences</title><description>Glukoz 6-fosfat dehidrogenaz (G6PD) enzimi kaz eritrositlerinden saflaştırıldı ve bazı karakteristik özellikleri araştırıldı. Saflaştırma prosedürü;' hemolizatın hazırlanması, amonyum sülfat çöktürmesi ve 2', 5'-ADP Sepharose 4B afınite kromatografisi şeklinde üç basamaktan ibarettir. Bu üç basamak neticesinde 36,2 EÜ/mg protein spesifik aktivitesine sahip olan enzim, % 68,79 verimle 3.892 kat saflaştırıldı ve enzim saflığının kontrol edilmesi için SDS-PAGE yapıldı. Enzim için optimum pH, stabil pH, optimum sıcaklık, molekül ağırlığı, NADP* için Km ve Vmax değerleri, G6-P için Km ve Vmax değerleri hesaplandı. Ayrıca ATP, ADP ve NADPH inhibitörleri için Lineweaver-Burk grafikleri çizilerek K, değerleri ve inhibisyon tipleri tespit edildi. Bu maddeler enzimi yarışmasız olarak inhibe etti.
Glucose 6-phosphate dehydrogenase (G6PD) was purified from goose erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of 3 steps: hemolysate preparation, ammonium sulfate precipitation, and 2', 5'-ADP Sepharose 4B affinity gel chromatography. Thanks to the 3 consecutive procedures, the enzyme, having a specific activity of 36.2 EU/mg protein, was purified for a yield of 68.79% and 3892 folds; to ascertain enzyme purity, SDS-PAGE was performed. Optimal pH, stable pH, optimal temperature, molecular weight, and Km and Vmax values for NADP* and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. In addition, K, values and inhibition type were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP and NADPH. These materials inhibited the enzyme in a noncompetitive manner.</description><subject>alyuvarlar</subject><subject>Animal Physiology and Biochemistry (Excluding Nutrition)</subject><subject>erythrocytes</subject><subject>geese</subject><subject>glucose-6-phosphate dehydrogenase</subject><subject>glukoz-6-fosfat dehidrogenaz</subject><subject>Hayvan Fizyolojisi ve Biyokimyası (Beslenme Dışında)</subject><subject>kaz</subject><subject>purification</subject><subject>saflaştırma</subject><issn>1300-0128</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid/><recordid>eNqFyr0OgjAUQOEOmog_b-DQFyAplqC70Tg6uJNabuEK9Da3ZeDt1cTd6QzfWYis0ErlqjicVmId40upgy4rlYn6PjE6tCYheUlOtsNkKYKs8tBRDJ1JIBvo5oapBW8-4phG2dJ3Ap4Tk50TRGl8I3v0kNDKwBSAE0LciqUzQ4Tdrxuxv14e51s-DaZ_4lgHxtHwXOtCHUv9h9_1iUDk</recordid><startdate>2003</startdate><enddate>2003</enddate><creator>YILMAZ, Hayrullah</creator><creator>KÜFREVİOĞLU, Ö.İrfan</creator><creator>ÇİFTÇİ, Mehmet</creator><creator>BAKAN, Ebubekir</creator><creator>BEYDEMİR, Şükrü</creator><general>TÜBİTAK</general><scope/></search><sort><creationdate>2003</creationdate><title>Purification of glucose 6-phosphate dehydrogenase from goose erytrocytes and kinetic properties</title><author>YILMAZ, Hayrullah ; KÜFREVİOĞLU, Ö.İrfan ; ÇİFTÇİ, Mehmet ; BAKAN, Ebubekir ; BEYDEMİR, Şükrü</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-ulakbim_primary_310743</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>alyuvarlar</topic><topic>Animal Physiology and Biochemistry (Excluding Nutrition)</topic><topic>erythrocytes</topic><topic>geese</topic><topic>glucose-6-phosphate dehydrogenase</topic><topic>glukoz-6-fosfat dehidrogenaz</topic><topic>Hayvan Fizyolojisi ve Biyokimyası (Beslenme Dışında)</topic><topic>kaz</topic><topic>purification</topic><topic>saflaştırma</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>YILMAZ, Hayrullah</creatorcontrib><creatorcontrib>KÜFREVİOĞLU, Ö.İrfan</creatorcontrib><creatorcontrib>ÇİFTÇİ, Mehmet</creatorcontrib><creatorcontrib>BAKAN, Ebubekir</creatorcontrib><creatorcontrib>BEYDEMİR, Şükrü</creatorcontrib><jtitle>Turkish journal of veterinary & animal sciences</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>YILMAZ, Hayrullah</au><au>KÜFREVİOĞLU, Ö.İrfan</au><au>ÇİFTÇİ, Mehmet</au><au>BAKAN, Ebubekir</au><au>BEYDEMİR, Şükrü</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification of glucose 6-phosphate dehydrogenase from goose erytrocytes and kinetic properties</atitle><jtitle>Turkish journal of veterinary & animal sciences</jtitle><date>2003</date><risdate>2003</risdate><volume>27</volume><issue>5</issue><spage>1179</spage><epage>1185</epage><pages>1179-1185</pages><issn>1300-0128</issn><abstract>Glukoz 6-fosfat dehidrogenaz (G6PD) enzimi kaz eritrositlerinden saflaştırıldı ve bazı karakteristik özellikleri araştırıldı. Saflaştırma prosedürü;' hemolizatın hazırlanması, amonyum sülfat çöktürmesi ve 2', 5'-ADP Sepharose 4B afınite kromatografisi şeklinde üç basamaktan ibarettir. Bu üç basamak neticesinde 36,2 EÜ/mg protein spesifik aktivitesine sahip olan enzim, % 68,79 verimle 3.892 kat saflaştırıldı ve enzim saflığının kontrol edilmesi için SDS-PAGE yapıldı. Enzim için optimum pH, stabil pH, optimum sıcaklık, molekül ağırlığı, NADP* için Km ve Vmax değerleri, G6-P için Km ve Vmax değerleri hesaplandı. Ayrıca ATP, ADP ve NADPH inhibitörleri için Lineweaver-Burk grafikleri çizilerek K, değerleri ve inhibisyon tipleri tespit edildi. Bu maddeler enzimi yarışmasız olarak inhibe etti.
Glucose 6-phosphate dehydrogenase (G6PD) was purified from goose erythrocytes and some characteristics of the enzyme were investigated. The purification procedure was composed of 3 steps: hemolysate preparation, ammonium sulfate precipitation, and 2', 5'-ADP Sepharose 4B affinity gel chromatography. Thanks to the 3 consecutive procedures, the enzyme, having a specific activity of 36.2 EU/mg protein, was purified for a yield of 68.79% and 3892 folds; to ascertain enzyme purity, SDS-PAGE was performed. Optimal pH, stable pH, optimal temperature, molecular weight, and Km and Vmax values for NADP* and glucose 6-phosphate (G6-P) substrates were also determined for the enzyme. In addition, K, values and inhibition type were determined by means of Lineweaver-Burk graphs obtained for such inhibitors as ATP, ADP and NADPH. These materials inhibited the enzyme in a noncompetitive manner.</abstract><pub>TÜBİTAK</pub><oa>free_for_read</oa></addata></record> |
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| subjects | alyuvarlar Animal Physiology and Biochemistry (Excluding Nutrition) erythrocytes geese glucose-6-phosphate dehydrogenase glukoz-6-fosfat dehidrogenaz Hayvan Fizyolojisi ve Biyokimyası (Beslenme Dışında) kaz purification saflaştırma |
| title | Purification of glucose 6-phosphate dehydrogenase from goose erytrocytes and kinetic properties |
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