SavedAdd your journal

Journals / Iğdır Üniversitesi Fen Bilimleri Enstitüsü Dergisi / 2026 / Cilt: 6 Sayı: 3

Comparision of Three Phase Partitioning (TPP) and Convantional Enzyme Purifcation; Thermodynamic Parameters of β-Galactosidase Purifed by TPP

Üçlü Faz Ayrımı (ÜFA) ile Geleneksel Enzim Saflaştırma Tekniğinin Karşılaştırılması; ÜFA ile Saflaştırılan β-Galaktosidazın Termodinamik Özellikleri

Yonca Duman
Pages
107–117
Publication Date
2016-09-30
DOI
—
🏛️Bibliographic metadata and full text access for this article are provided via DergiPark.
DergiPark Record ↗
✨

AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

In this study thermodynamic parameters of β-Galactosidase as ΔG # , 62.21 kJ mol -1 ; Δ #E-T , -13.56 kJ mol -1 ; Δ #E-S , 0.25 kJ mol -1 ; ΔH # , 26.06 kJ mol -1 ; ΔS # , -0.12 kJ mol -1 K -1 which was purifed by TPP

📌 3 Key Takeaways

🎯
Objective & Hypothesis:Enzymes have many industrial applications.
📊
Key Findings & Data:Unfotunately purifcation cost restricts of using area of this importatant biocataliyst.
💡
Conclusion & Impact:In this study thermodynamic parameters of β-Galactosidase as ΔG # , 62.21 kJ mol -1 ; Δ #E-T , -13.56 kJ mol -1 ; Δ #E-S , 0.25 kJ mol -1 ; ΔH # , 26.06 kJ mol -1 ; ΔS # , -0.12 kJ mol -1 K -1 which was purifed by TPP
🔬Design:Araştırma Makalesi
📍Scope:Comprehensive Sample
⚙️Variables:β-Galaktosidaz · saflaştırma · termodinamik özellikler

Abstract

Enzymes have many industrial applications. Unfotunately purifcation cost restricts of using area of this importatant biocataliyst. Recent years studies on enzyme purifcation is about reducing of purifcation cost by reducing steps and time saving. Three phase partitioning (TPP) is important purifcation method. TPP uses tbutanol and ammonium sulfate to precipitate enzymes and proteins from aqueous solution. The simplicity of the method combined with its rapid result make this a popular choice in large scale protein purifcation. In this study thermodynamic parameters of β-Galactosidase as ΔG # , 62.21 kJ mol -1 ; Δ #E-T , -13.56 kJ mol -1 ; Δ #E-S , 0.25 kJ mol -1 ; ΔH # , 26.06 kJ mol -1 ; ΔS # , -0.12 kJ mol -1 K -1 which was purifed by TPP

Özet

Enzimlerin yaygın endüstriyel kullanımları mevcuttur. Ancak bu önemli biyokatalizörlerin saflaştırma maliyetleri kullanım alanlarını kısıtlamaktadır. Son yıllarda yapılan çalışmalar enzimlerin saflaştırma basamaklarını düşürerek maliyet ve zaman tasarrufu yapmak yönündedir. Bu amaçla Üçlü faz yöntemi (ÜFA) ile enzim saflaştırması önemli bir yöntemdir. Hedef enzim Üçlü faz yönteminde t-bütanol ve amonyum sülfat ile kullanılarak enzim çözeltisi tuzlu ara fazda çöktürülür. Metodun basitliğinin yanı sıra hızlı sonuç vermesi ve düşük maliyeti endüstriyel kullanımını cazip hale getirmektedir. Bu çalışmada enzimlerin geleneksel yöntemler ile ÜFA saflaştırılma protokolleri karşılaştırılarak yöntemlerin birbiri üzerine üstünlükleri tartışıldı. Aynı zamanda ÜFA Yöntemi ile saflaştırılan β-galaktosidaz enziminin termodinamik özellikleri ve ΔG # , 62.21 kJ mol -1 ; Δ #E-T , -13.56 kJ mol -1 ; Δ #E-S , 0.25 kJ mol -1 ; ΔH # , 26.06 kJ mol -1 ; ΔS # , -0.12 kJ mol -1 K -1 olarak hesaplandı.

🔖

Cite This Article

Duman, Y. (2016). Comparision of Three Phase Partitioning (TPP) and Convantional Enzyme Purifcation; Thermodynamic Parameters of β-Galactosidase Purifed by TPP. Iğdır Üniversitesi Fen Bilimleri Enstitüsü Dergisi, Cilt: 6 Sayı: 3, 107–117. https://ajindex.com/articles/1273119

📄 Full Text PDF Viewer

📄

PDF görüntüleyici hazırlanıyor...

References (30)

  1. Altman S, Groseclose C, Ma JX, Hamamdzic D, Vrinda vanam NS., Middaugh LD., Parratto NP, Salle FR, 1997. Expression of beta-galactosidase in mouse brain: utilization of a novel nonreplicative Sindbis virus vector as a neuronal gene delivery system. Gene Therapy, 4: 815-822.
  2. Angal S, Harris E, 1990. Protein Purifcation Applications, Second Edition, IRL Pres at Oxford University Pres, UK. 317p.
  3. Atav R, Yurdakul A, Arabacı A, 2009. Tekstil boyacılığında kullanılan tuzların özellikleri ve kullanım amaçları. Tekstil Teknolojileri Elektronik Dergisi, 3: 71-80.
  4. Ay T, Bingöl G, Binici MS, Diler SB, Cansaran A, Demirağ MK, Dere E, Doğan N, Doğan B, Ekici N, Gülnaz O, Kanlı A, Olguner E, Polat F, Sözbilen M, Şimşekli Y, Yıldırım C, 2010. Biyolojide Özel Konular, Üçüncü Basım, Pegem Akademi, TÜRKİYE, 376s.
  5. Bollag DM, Rozycki MD, Edelstein SJ, Protein Methods, 1994. Second Edition, Wiley-Liss, New York, 415p.
  6. Chandler V, Donovan S, Goodwin W, Sprague S, Stiefbold F, 1998. Enzyme kinetics. Proceeding of the 19th Workshop/ Conference of the Association for Biology Laboratory Education (ABLE), 19: 81-97.
  7. Dennison C, Lovrein R, 1997. Three phase partitioning: concentration and purifcation of proteins. Protein Exp. Purif., 11: 149-161.
  8. Duman YA, 2008. Bacillus clausii alkalen proteazının su ile karışabilen organik çözücüler varlığında kinetik ve termodinamik özelliklerinin incelenmesi. Kocaeli Üniversitesi, Fen Bilimleri Enstitüsü (Basılmış), Doktora Tezi, 308s.
  9. Duman YA, Kaya E, 2013a. Three-Phase Partitioning as a Rapid and Easy Method for the Purifcation and Recovery of Catalase from Sweet Potato Tubers (Solanum tuberosum). Appl Biochem Biotechnol, 170:1119–1126.
  10. Duman YA, Kaya E, 2014. Purifcation and recovery of invertase from potato tubers (Solanum tuberosum) by three phase partitioning and determination of kinetic properties of purifed enzyme. Türk Biyokimya Dergisi [Turkish Journal of Biochemistry–Turk J Biochem] 39(4): 443–448.
  11. Duman YD, Kaya E, 2013b. Purifcation, recovery, and characterization of chick pea (Cicer arietinum) β-galactosidase in single step by three phase partitioning as a rapid and easy technique. Protein Expression and Purifcation, 91: 155-160.
  12. Erarslan A, Kazan D, Denizci AA, Öztürk DC, Karahan N, 2008. Tubitak Gen Mühendisliği ve Biyoteknoloji Araştırma Enstitüsü Enzim Saflaştırmada Temel Yöntemler VIII. Uygulamalı Eğitim Kursu Kitabı, 175s.
  13. Garman J, Coolbear T, Smart J, 1996. The effect of cations on the hydrolysis of lactose and the transferase reactions catalysed by β-galactosidase from six strains of lactic acid bacteria. Appl. Microbiol.Biotechnol, 46: 22-37.
  14. Greenberg NA., Mahoney RR, 1982. Production and characterization of β-galactosidase from Streptococcus thermophilus. J. Food Sci, 47: 1824-1828.
  15. Hoyoux A, Jennes I, Dubois P, Genicot S, Dubail F, François JM, Baise E, Feller G, Gerday C, 2001. Cold-Adapted β-Galactosidase from the Antarctic Psychrophile Pseudoalteromonas haloplanktis. Appl. Environ. Microbiol, 67: 1529-1535.
  16. Jurado E, Camacho F, Luzon G, Vicaria JM, 2002. A new kinetic model proposed for enzymatic hydrolysis of lactose by a β-galactosidase from Kluyveromyces fragilis. 31: 300-309.
  17. Kang I K, Suh SG, Gross KC, Byun JK, 1994. N – terminal amino acid sequence of persimmon fruit beta-Galactosidase. Plant Physiol, 105: 975-979.
  18. Kishore D, Kayastha AM, 2012. A β-galactosidase from chick pea (Cicer Arietinum) seeds: its purifcation, biochemical properties and industrial applications. Food Chem, 134: 1113- 1122.
  19. Lee D H, Kang SG., Suh SG, Byun J K, 2003. Purifcation and characterization of a beta-galactosidase from peach (Prunus persica). Molecules and Cells, 15: 68-74.
  20. Macfarlane GT, Steed H, Macfarlane S, 2008. Bacterial metabolism and health-related effects of galacto-oligosaccharides and other prebiotics. J. Appl. Microbiology, 104: 305-344.
  21. McBean LD, Miller GD, 1998. Allaying fears and fallacies about lactose intolerance. Journal of the American Dietetic Association, 98: 671-676.
  22. Nostro PL, Ninham BW, 2012. Hofmeister phenomena: an update on ion specifcity in biology. Chem. Rev, 112: 2286-2322.
  23. Okada S, O’Brien JS, 1968. Generalized gangliosidosis: Beta - galactosidase defciency. Science, 160: 1002-1004.
  24. Olichon A, Schweizer D, Muyldermans S, Marco A, 2010. Heating as a rapid purifcation method for recovering correctly-folded thermotolerant VH and VHH domains. BMC Biotechnol, DOI: 10.1186/1472-6750-7-7.
  25. Pal A, Lobo M, Khanum F, 2013. Extraction, Purifcation and thermodynamic characterization of almond (Amygdalus communis) β-galactosidase for the preparation of delactosed milk. Food Technol. Biotechnol, 51: 53-61.
  26. Panesar R, Panesar PS, Singh RS, Kennedy JF, Puri M, 2011. Hydrolysis of milk lactose in a packed bed reactor system using immobilised yeast cells. J. Chem. Technol. Biotechnol. 86: 42-46.
  27. Puri M, Gupta S, Pahuja P, Kaur A, Kanwar J E, Kennedy J F, 2010. Cell Disruption Optimization and Covalent Immobilization of β-D-Galactosidase from Kluyveromyces marxianus YW-1 for Lactose Hydrolysis in Milk. Appl. Biochem. Biotechnol, 160: 98-108.
  28. Scopes RK, 1984. Protein Purifcation, Second Edition, SpringerVerlag, New York, 345s. Tüzün C, 1991. Biyokimya, Birinci Basım, Palme Yayınevi, Ankara, 486s.
  29. Uyanık A, 2008. Beta - galaktosidaz enziminin mikrobiyal hücrelerden izolasyonu ve karakterizasyonu. Ankara Üniversitesi, Fen Bilimleri Enstitüsü (Basılmış), Yüksek Lisans Tezi, 72s.
  30. Vermaa ML, Barrowa C J., Kennedy J K, Puri M, 2012. Immobilization of β-D- galactosidase from Kluyveromyces lactis on functionalised silicon dioxide nanoparticles: characterization and lactose hydrolysis. Int. J. Biol. Macromol, 50: 432-437.