Bacillus pumilus Y7 alkalen proteaz enziminin bentonit ile non-kovalent immobilizasyonu
dc.contributor.advisor | Duman, Yonca | |
dc.contributor.author | Cengiz Şekerli, Necla | |
dc.date.accessioned | 2020-12-29T12:54:16Z | |
dc.date.available | 2020-12-29T12:54:16Z | |
dc.date.submitted | 2018 | |
dc.date.issued | 2018-12-04 | |
dc.identifier.uri | https://acikbilim.yok.gov.tr/handle/20.500.12812/415228 | |
dc.description.abstract | Proteazlar, proteinlerde bulunan peptit bağlarını katalizleyen enzimlerdir. Doğada yaygın olarak bulunurlar. Bitkisel, hayvansal ve mikrobiyal kaynaklardan izole edilirler. Bu çalışmamızda Bacillus pumilus Y7'den elde edilen alkalen proteaz enziminin, bentonit ile non-kovalent immobilizasyonu gerçekleştirildi. Serbest enzim ve immobilize enzimin kinetik ve termodinamik parametreleri incelendi. SDS-PAGE analizi ile enzimin molekül ağırlığı 15 kDa olarak bulundu, native-PAGE ve zimogram analizi ile de desteklendi. Serbest enzim için optimum pH ve sıcaklık sırasıyla 10 ve 35 ℃, immobilize enzim için optimum pH ve sıcaklık sırasıyla 6 ve 32 ℃ olarak bulundu. Arrhenius aktivasyon enerjileri (Ea) serbest enzim ve immobilize enzim için sırasıyla 15,28 kJ/mol ve 102,4 kJ/mol olarak hesaplandı. 30 ℃'de pH 8'de her iki enzim için Michaelis-Menten sabiti (Km) ve maksimum hızı (Vm) serbest enzim için sırasıyla 6,05×10-6 M ve 20,16 U/mL/min, immobilize enzim için sırasıyla 27,3×10-6 M ve 34,13 U/mL/min olarak bulundu. Enzimlerin turnover sayısı (kcat) ve katalitik performansı (kcat/Km) serbest enzim için sırasıyla 24,39 dk-1 ve 4×106 dk-1M-1, immobilize enzim için sırasıyla 180,4 dk-1 ve 6,6×106 dk-1M-1 olarak bulundu. Serbest enzimin termodinamik parametreleri; ∆G#: 66,21 kJ/mol; ∆G#E-T: -38,31 kJ/mol; ∆G#ES: -30,27 kJ/mol; ∆H#:13,3 kJ/mol; ∆S#: -0,17 kJ/molK ve immobilize enzimin termodinamik parametreleri ∆G#: 61,15 kJ/mol; ∆G#E-T: -56,22 kJ/mol; ∆G#ES: 26,45 kJ/mol; ∆H#: 99,52 kJ/mol; ∆S#: 0,12 kJ/molK olarak hesaplandı. | |
dc.description.abstract | Proteases are enzymes that catalyse the peptide bonds in proteins. They are widely found in nature. They are isolated from plant, animal and microbial sources. In this study, non-covalent immobilization of alkaline protease enzyme from Bacillus pumilus Y7 with bentonite was performed. Kinetic and thermodynamic parameters of free enzyme and immobilized enzyme have been studied. The molecular weight of the enzyme was found 15 kDa by SDS-PAGE analysis and was also supported by native-PAGE and zymogram analysis. Optimum pH for free enzyme and immobilized enzyme were 10.0 and 6.0, respectively and optimum temperature were determined for free enzyme and immobilized enzyme 35 °C and 32 °C, respectively. The Arrhenius activation energies (Ea) were calculated as 15.28 kJ/mole and 102.4 kJ/mole for the free enzyme and the immobilized enzyme, respectively. The Michaelis-Menten constant (Km) and the maximum velocity (Vm) at pH 8, 30 °C for the free enzyme were 6.05×10-6 M and 20.16 U/mL/min respectively, and for the immobilized enzyme 27.3×10-6 M and 34.13 U/mL/min, respectively. The turnover number (kcat) and catalytic performance (kcat / Km) of the enzymes were found to be 24.39 min-1 and 4x106 min-1 for the free enzyme, 180.4 min-1 and 6.6x106 min-1 for the immobilized enzyme in order. Thermodynamic parameters of free enzyme were calculated as; ∆G#: 66.21 kJ/mole; ∆G#E-T: -38.31 kJ/mole; ∆G#ES: -30.27 kJ/mole; ∆H# : 13.3 kJ/mole; ∆S#: -0.17 kJ/moleK and the thermodynamic parameters of immobilized enzyme were calculated as ∆G#: 61.15 kJ/mole; ∆G#E-T: -56.22 kJ/mole; ∆G#ES: -26.45 kJ/mole; ∆H#: 99.52 kJ/mole; ∆S#: 0.12 kJ/moleK. | en_US |
dc.language | Turkish | |
dc.language.iso | tr | |
dc.rights | info:eu-repo/semantics/openAccess | |
dc.rights | Attribution 4.0 United States | tr_TR |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | |
dc.subject | Biyokimya | tr_TR |
dc.subject | Biochemistry | en_US |
dc.subject | Kimya | tr_TR |
dc.subject | Chemistry | en_US |
dc.title | Bacillus pumilus Y7 alkalen proteaz enziminin bentonit ile non-kovalent immobilizasyonu | |
dc.title.alternative | Non-covalent immobilization of Bacillus pumilus Y7 alkaline protease with bentonite | |
dc.type | masterThesis | |
dc.date.updated | 2018-12-04 | |
dc.contributor.department | Kimya Anabilim Dalı | |
dc.identifier.yokid | 10201647 | |
dc.publisher.institute | Fen Bilimleri Enstitüsü | |
dc.publisher.university | KOCAELİ ÜNİVERSİTESİ | |
dc.identifier.thesisid | 521698 | |
dc.description.pages | 91 | |
dc.publisher.discipline | Diğer |