Separation and purification of lipase using Cu nanoparticle embedded poly HEMA-MATrp cryogels

Year 2014, , 43 - 50, 31.12.2014

Kadir Erol Kazim Kose Dursun Ali Kose Gulcin Alp Avci Lokman Uzun

https://doi.org/10.17350/HJSE19030000007

Cited By: 3

Abstract

Quality and efficiency of techniques to be used for separation and purification lipase enzymes are commercially important enzyme. Among such techniques, adsorption methods are highly preferred. Cryogels have been quite extensively used as the adsorbents due to their macropores and interconnected flow channels. In this study, adsorption of lipase enzyme onto copper nanoparticles embedded poly 2-hydroxyethyl methacrylate-N-methacryloyl-L-tryptophan , poly HEMA-MATrp cryogels was studies for conditions with varying pH, interaction time, lipase enzyme initial concentration, temperature and ionic strength. Maximum lipase enzyme adsorption capacity of cryogels was determined as 183.6 mg/g. Fourier transform infrared spectrometer FTIR and scanning electron microscopy SEM were used for characterization of cryogels. At the end of the adsorption process, in order to be sure that the purity of lipase enzyme desorbed from cryogels, SDS-PAGE analyses were performed and molecular weight of the lipase enzyme was determined as 58 kDa. Adsorption characteristic of cryogels were determined according to the results of Langmuir and Freundlich adsorption isotherm models. As a result of calculation run for adsorption isotherm models, Langmuir isotherm model was determined to be more appropriate.

Keywords

Lipase, Cupper, Cryogel, Adsorption.

References

• Kapoor M, Gupta MN. Lipase promiscuity and its biochemical applications. Process Biochemistry 47 (2012) 555–569.
• Mendes AA, Oliveira PC, Castro HF, Properties and biotechnological applications of porcine pancreatic lipase. Journal of Molecular Catalysis B: Enzymatic 78 (2012) 119– 134.
• Fernandez-Lafuente R. Lipase from Thermomyces lanuginosus: uses and prospects as an industrial biocatalyst. Journal of Molecular Catalysis B: Enzymatic 62 (2010) 197– 212. Sharma D, Sharma B, Shukla AK. Biotechnological approach of microbial lipase: a review. Biotechnology 10 (2011) 23–40.
• Hasan F, Shah AA, Hameed A. Industrial applications of microbial lipases. Enzyme and Microbial Technology 39 (2006) 235–251.
• Chesterfield DM, Rogers PL, Al-Zaini EO, Adesina AA, Production of biodiesel via ethanolysis of waste cooking oil using immobilised lipase. Chemical Engineering Journal 207–208 (2012) 701–710.
• Olivares-Carrillo P, Quesada-Medina J, de los Ríos AP, Hernández-Fernández FJ, Estimation of critical properties of reaction mixtures obtained in different reaction conditions during the synthesis of biodiesel with supercritical methanol from soybean oil. Chemical Engineering Journal 241 (2014) 418–432.
• Umare SS, Chandure AS. Synthesis, characterization and biodegradation studies of poly(ester urethane)s. Chemical Engineering Journal 142 (2008) 65–77.
• Wang C, Dong XY, Jiang Z, Sun Y. Enhanced adsorption capacity of cryogel bed by incorporating polymeric resin particles. Journal of Chromatography A 1272 (2013) 20-25.
• Zhao W, Zhang S, Lu M, Shen S, Yun J, Yao K, et al. Immiscible liquid slug flow characteristics in the generation of aqueous drops within a rectangular microchannel for preparation of poly(2-hydroxyethylmethacrylate) cryogel beads. Chemical Engineering Research and Design 2014. http://dx.doi. org/10.1016/j.Cherd.2014.01.012 [in press].
• Uzun L, Armutcu C, Biçen O, Ersöz A, Say R, Denizli A.
• Simultaneous depletion of immunoglobulin G and albumin from human plasma using novel monolithic cryogel columns. Colloids and Surfaces B: Biointerfaces 112 (2013) 1-8.
• Yuna J, Jespersen GR, Kirsebom H, Gustavsson PE, Mattiasson B, Galaeva IY. An improved capillary model for describing the microstructure characteristics, fluid hydrodynamics and breakthrough performance of proteins in cryogel beds. Journal of Chromatography A 1218 (2011) 5487-5497.
• Eichhorn T, Ivanov AE, Dainiak MB, Leistner A, Linsberger
• I, Jungvid H, et al. Macroporous composite cryogels with embedded polystyrene divinylbenzene microparticles for the adsorption of toxic metabolites from blood. Journal of Chemistry ttp://dx.doi.org/10.1155/2013/348412.
• Hajizadeh S, Xu C, Kirsebom H, Ye L, Mattiasson B. Cryogelation of molecularly imprinted nanoparticles: a macroporous structure as affinity chromatography column for removal of b-blockers from complex samples. Journal of Chromatography A 1274 (2013) 6-12.
• Plieva FM, Kirsebom H, Mattiasson B. Preparation of
• macroporous cryostructurated gel monoliths, their characterization and main applications. Separation Scientific 34 (2011) 2164-2172.
• Plieva FM, Galaev IY, Noppe W, Mattiasson B. Cryogel applications in microbiology. Trends in Microbiology 16 (2008) 543-551.
• Yilmaz F, Bereli N, Yavuz H, Denizli A. Supermacroporous hydrophobic affinity cryogels for protein chromatography. Biochemical Engineering Journal 43 (2009) 272–279.
• Zhang W, Luan C, Yang Z, Liu X, Zhang D, Yang S. Preparation and optical properties of Cu 2O hollow microsphere film and hollow nanosphere powder via a simple liquid reduction approach. Applied Surface Science 253 (2007) 6063-6067.
• Qing-Ming L, Yasunami T, Kuruda K, Okido M. Preparation
• of Cu nanoparticles with ascorbic acid by aqueous solution reduction method. Transactions of Nonferrous Metals Society of China 22 (2012) 2198-2203.
• Asliyuce S, Uzun L, Say R, Denizli A. Immunoglobulin G recognition with Fab fragments imprinted monolithic cryogels: Evaluation of the effects of metal-ion assisted- coordination of template molecule. Reactive and Functional Polymers. 73 (2013) 813-820.
• Langmuir I. The adsorption of gas on plane surfaces of glass, mica and platinum. Journal of the American Chemical Society 40 (1918) 1361–1370.
• Freundlich HMF, Uber die adsorption in losungen. Zeitschrift für. Physikalische Chemie. 57(1906) 385–471.