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T.C. Selçuk Ünġversġtesġ Fen Bġlġmlerġ Enstġtüsü Tuzlu

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T.C. Selçuk Ünġversġtesġ Fen Bġlġmlerġ Enstġtüsü Tuzlu T.C. SELÇUK ÜNĠVERSĠTESĠ FEN BĠLĠMLERĠ ENSTĠTÜSÜ TUZLU TOPRAKLARDA KATALAZ ENZĠMĠNĠN AKTĠVĠTESĠ VE KĠNETĠĞĠ Emine YILDIRIM YÜKSEK LĠSANS TEZĠ TOPRAK BĠLĠMĠ VE BĠTKĠ BESLEME ANABĠLĠM DALI KONYA, 2010 ÖZET YÜKSEK LĠSANS TEZĠ TUZLU TOPRAKLARDA KATALAZ ENZĠMĠNĠN AKTĠVĠTESĠ VE KĠNETĠĞĠ Emine YILDIRIM Selçuk Üniversitesi Fen Bilimleri Enstitüsü Toprak Bilimi ve Bitki Besleme Anabilim Dalı DanıĢman: Yrd. Doç. Dr. Fariz MĠKAĠLSOY 2010, Sayfa: 72 Jüri: Yrd. Doç. Dr. Fariz MĠKAĠLSOY Prof. Dr. Nizamettin ÇĠFTÇĠ Doç. Dr. Refik UYANÖZ Bu araĢtırmada, Tuz gölü çevresi tarım dıĢı arazilerden alınan 3 toprak örneğinde çalıĢılmıĢtır. Gazometrik metod kullanılarak katalaz enziminin aktivitesi tayin edilerek kinetik parametreleri hesaplanmıĢtır. Fiziksel, kimyasal özellikleri ve % tuz oranı farklı toprakların katalaz enzim analizi 20+1 oC laboratuar koĢullarında değiĢik substrat konsantrasyonlarda (% 3, % 6, % 9, % 12, % 15, % 18, %2 1, % 24, % 27, % 30 H2O2) yürütülmüĢtür. Bu analizde ürün olarak açığa çıkan O2‟nin zamana göre değiĢimi (20, 40, 60, 80,….300 sn) kararlı hale gelmesine kadar devam edilmiĢtir. Katalaz enzim aktivitesi (υ) ve kinetik parametreleri (υ0, Vmax ve Km) her toprak için ayrı ayrı yapılmıĢtır. Sonuçlara göre, tuz konsantrasyonu yüksek toprakta katalaz enziminin kinetik parametresi olan Km‟nin değeri yüksek bulunmuĢtur. Vmax değeri ise en düĢük olarak bulunmuĢtur. Ayrıca reaksiyon hızının % 24 substrat konsantrasyonunda artıĢ gösterdiği ve daha sonra değiĢmediği tesbit edildi. Bu metod toprakta katalaz enzim aktivitesinin tesbitinde kullanılabilir. Anahtar Kelimeler: Toprak, enzim aktivitesi, katalaz, kinetik parametreler, tuz i ABSTRACT MASTER THESĠS ACTIVITY AND KINETICS OF CATALASE ENZYME IN SALINE SOILS Emine YILDIRIM Selçuk University Graduate School of Natural and Applied Sciences Department of Soil Science and Plant Nutrition Supervisor: Yrd. Doç. Dr. Fariz MĠKAĠLSOY 2010, Pages: 72 Jury: Yrd. Doç. Dr. Fariz MĠKAĠLSOY Prof. Dr. Nizamettin ÇĠFTÇĠ Doç. Dr. Refik UYANÖZ In this study, 3 soil samples taken from the non-agricultural land around the Salt Lake were examined. The kinetics parameters were calculated by determining the activity of catalase enzyme by using the Gasometric method. The catalase enzyme analysis of samples having different physical and chemical properties and % salt ratios was conducted at temperatures of 20+1 oC in laboratory conditions and different substrate concentrations (% 3, % 6 ,% 9,% 12, % 15 ,% 18 ,% 21 % 24 ,% 27, % 30 H2O2). In this analysis was continued until the amount of O2 brought out as product with respect to time (20, 40, 60, 80, ... .300 sec) became stable. Catalase enzyme activity (υ) and kinetic parameters (υ0, Vmax and Km) were made separately for each sample. According to results, the value of Km , the kinetic parameter of the catalase enzyme, was maximum in soils having high concentration of salt. Vmax values were found to be low. In addition, it was determined that the reaction rate increased at % 24 substrate concentration and then did not change. This method can be used to determine the catalase enzyme activity in soil. Key Words : Soil, enzyme activity, catalase, kinematics parameters, salt ii TEġEKKÜR Bu araĢtırmanın yüksek lisans tezi olarak planlanıp, yürütülmesi ve sonuçlarının değerlendirilmesinde daima yardım ve ilgisini gördüğüm danıĢman hocam Sayın Yrd. Doç. Dr. Fariz MĠKAĠLSOY‟a, çalıĢmalarım esnasında yardımlarını esirgemeyen Doç. Dr. Refik UYANÖZ‟e ve ve laboratuar çalıĢmalarında yardımlarını gördüğüm ArĢ. Gör. Ümmühan KARACA, ArĢ. Gör. Fatma GÖKMEN ve Uzman Emel KARAASLAN‟ a olumlu desteklerinden dolayı aileme teĢekkürü bir borç bilirim. KONYA, 2010 Emine YILDIRIM iii ĠÇĠNDEKĠLER Sayfa No ÖZET …………………………………………………………………………… i ABSTRACT ........................................................................................................ ii TEġEKKÜR ....................................................................................................... iii ĠÇĠNDEKĠLER .................................................................................................. iv ÇĠZELGELER DĠZĠNĠ ..................................................................................... vii ġEKĠLLER DĠZĠNĠ ........................................................................................... x GRAFĠKLER DĠZĠNĠ........................................................................................ xi 1. GĠRĠġ .......................................................................................................... 1 2. KAYNAK ÇALIġMALARI ..................................................................... 5 2.1. Topraktaki Enzim Aktivitesi Ġle Ġlgili ÇalıĢmalar…………..……… 5 2.2. Topraktaki Katalaz Enzim Aktivitesi Ġle Ġlgili ÇalıĢmalar…….……. 11 2.3. Topraktaki Enzim Aktivitesinin Kinetik Parametreler Üzerine Etkisi Ġle Ġlgili ÇalıĢmalar…………………………………………………. 18 2.4 Tuzlu Toprak ve Tuzlu Topraklarda Enzim Aktivitesi Ġle Ġlgili ÇalıĢmalar…………………………………………………………… 23 3. MATERYAL VE METOD…………………………………………..….. 27 3.1. Materyal………………………………………………………...…… 27 3.1.1. Toprak Örneklerinin Alındığı Bölge Hakkında Genel Bilgiler………………………………………………….….. 27 3.1.2. Toprak Örneklerinin Alındığı Arazinin Toprak Özellikleri. 28 3.1.3. Toprak Örneklerinin Alındığı Bölgenin Ġklim Özellikleri… 30 3.1.4. Toprak Örneklerinin Alınması Ve Analize Hazırlanması.… 32 3.2. Metod………………………………………………………….…… 32 . 3.2.1 Toprak Örneklerinde Yapılan Fiziksel Ve Kimyasal Analizler………………………………………….….…… 32 3.2.2. Toprak Örneklerinin Biyolojik Analizleri……….……..… 33 3.2.3. Ġstatistiksel Analizler………………………….……….…. 39 4. ARAġTIRMA SONUÇLARI VE TARTIġMA………………….….. 40 4.1. AraĢtırma Topraklarının Bazı Fiziksel ve Kimyasal Özellikleri……. 40 4.2. Katalaz Enzim Aktivitesi Sonuçları…………………….………..… 45 4.3. Kinetik Parametrelerin Hesaplanması (Vmax ve )…………..….. 55 iv 5. SONUÇ ÖNERĠLER…………………………………………..……….... 59 6. KAYNAKLAR …………………………………………………………… 61 v ÇĠZELGELER DĠZĠNĠ Sayfa No 1. Çizelge 2.1. Tarla Topraklarının Enzim Aktiviteleri seviyeleri (Hofmann ve ark., 1966)………………………………………………..……. 7 2. Çizelge 2.2. Bakteri Sayısı Ġle Sakkaroz Aktivitesi Arasındaki ĠliĢki (Ergene, 1970)…………………………………………………. 8 3. Çizelge 2.3. Tarla ve Çayır Topraklarında ÇeĢitli derinliklerde Enzim Aktivitesi (Ergene,1970)…………………………………...….. 9 4. Çizelge 2.4. Türkiye topraklarının tuzluluk derecesi ve alanları………..….. 25 5. Çizelge 3.1. Havzaya Ait Bazı Meteorolojik Veriler Uzun Yıllar Ġçinde GerçekleĢen Ortalama Değerler (1975-2008).....................…… 31 6. Çizelge 3.2. t sn aralıklarında O2 ml olarak ölçüm değerleri………………. 36 7. Çizelge 3.3. BaĢlangıç hızının hesaplanması……………………………….. 37 8. Çizelge 4.1. AraĢtırma Alanından Alınan Toprakların Fiziksel özellikleri… 42 9. Çizelge 4.2. AraĢtırma Alanından Alınan Toprakların Kimyasal özellikleri.. 43 10. Çizelge 4.3. Toprakların Ekstrakte Edilebilir Ġyon Değerleri………………. 44 11. Çizelge 4.4. Laboratuvar KoĢullarında 1 Nolu Topraktaki % H2O2 Konsantrasyona Göre O2 ÇıkıĢ Miktarı……………………… 46 12. Çizelge 4.5. Laboratuvar KoĢullarında 2 Nolu Topraktaki % H2O2 Konsantrasyona Göre O2 ÇıkıĢ Miktarı ………………….…… 47 13. Çizelge 4.6. Laboratuvar KoĢullarında 3 Nolu Topraktaki % H2O2 Konsantrasyona Göre O2 ÇıkıĢ Miktarı …………………….… 48 14. Çizelge 4.7. Toprakların % [S] konsantrasyondaki hız (υ) değerleri ………. 54 15 Çizelge 4.8. Toprakların 1/ [S] ve 1/υ değerleri…………………………...... 56 16. Çizelge 4.9. AraĢtırma Topraklarının Kinetik Parametrelerinin ve Ġstatistiki Değerler……………………………………………………….. 57 vi ġEKĠL VE RESĠM DĠZĠNĠ Sayfa No 1. ġekil 2.1. H2O2 (Hidrojenperoksit) in Farklı Hallerde Atom Bağları 12 Arasındaki Açılar……………………………………………… 2. ġekil 3.1. Tuz gölü havzası ………………….…………………………… 28 3. Resim 3.1. AraĢtırma Topraklarının Alındığı Alan ..…………..…..……… 30 4. ġekil 3.2. Kalsimetre Aletin GörünüĢü………………………………..…. 34 5. Resim 3.2. Katalaz Enzim Analiz Seti......................................................... 35 6. ġekil 3.3. Farklı (3, 6, 9, … , 30 %) H2O2 için reaksiyon eğrileri……….. 36 7. ġekil 3.4. Bir enzimsel reaksiyonda substrat (S) konsatrasyonu ile baĢlangıç hız (v0) arasındaki iliĢkiyi gösteren doyma eğrisi….. 38 8. ġekil 3.5. Lineweaver-Burk Yönteminin OluĢturduğu Eğri……………… 39 vii GRAFĠK DĠZĠNĠ Sayfa No 1. Grafik 4.1. Toprakların % 3 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı.. 49 2. Grafik 4.2. Toprakların % 6 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı.. 49 3. Grafik 4.3. Toprakların % 9 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı.. 50 4. Grafik 4.4. Toprakların % 12 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı. 50 5. Grafik 4.5. Toprakların % 15 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı. 51 6. Grafik 4.6. Toprakların % 18 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı. 51 7. Grafik 4.7. Toprakların % 21 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı. 52 8. Grafik 4.8. Toprakların % 24 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı. 52 10. Grafik 4.9. Toprakların % 27 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı. 53 11. Grafik 4.10. Toprakların % 30 H2O2 konsantrasyonundaki O2 çıkıĢ miktarı. 53 12. Grafik 4.11. Enzimatik reaksiyon hızına substrat konsantrasyon etkisi .… 54 13. Grafik 4.12. Lineweaver-Burk Yöntemi, 1/[S] ve 1/υ değerleri.…………. 56 viii 1.GĠRĠġ Toprak kalitesinin değerlendirilmesinde; son yıllarda yapılan çalıĢmalarda sadece ürün verimi, arazi bozulması, erozyon ya da fiziksel ve kimyasal toprak faktörleri üzerine odaklanmak yerine toprakların fiziksel, kimyasal ve biyolojik özelliklerin hangi ölçekte toprak kalitesini belirlediği üzerinde durulmaktadır (Özulu ve ark. 2006). Tarımsal üretimde birim alandan
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