Abstract
Although ultrasound has been used to accelerate many enzymatic reactions, the low stability of enzymes in such a system still remains a critical issue, limiting its industrial application. Here, we have reported that polyethylene glycol (PEG) pretreatment stabilized Chromobacterium viscosum lipase (CVL) in ultrasound-assisted water-isooctane emulsion. PEGs of different molecular weights and concentrations were used to pretreat CVL, and the pretreated lipase activities for olive oil hydrolysis were investigated at different ultrasonic powers. The best result was attained with PEG400 at 100 mg/ml for a lipase concentration of 0.02 mg/ml and an ultrasonic power of 106 W. The half-life time of PEG400-treated lipase at 106 W was 54 min, a 27-fold higher than that attained using untreated lipase. Circular dichroism (CD) spectra suggested that PEG increased the rigidity of CVL structure, which favored the lipase stability against ultrasound inactivation. These results have important implications for the exploitation of ultrasound in biocatalytic process.
Similar content being viewed by others
References
Khemelnitsky, Y. L., Levashov, A. V., Klyachko, N. L., & Martinek, K. (1988). Engineering biocatalytic systems in organic media with low water content. Enzyme Microbial Technology, 10, 710–724.
Hossain, M. J., Takeyama, T., Hayashi, Y., Kawanishi, T., Shimizu, N., & Nakamura, R. (1999). Enzymatic activity of Chromobacterium viscosum lipase in an AOT/Tween 85 mixed reverse micellar system. Journal of Chemical Technology andBiotechnology.,74, 423–428.
Talukder, M. M. R., Hayashi, Y., Takeyama, T., Zaman, M. M., Wu, J. C., Kawanishi, T., & Shimizu, N. (2003). Activity and stability of Chromobacterium viscosum lipase in modified AOT reverse micelles. Journal of Molecular Catalysis B-Enzymatic, 22, 203–209.
Hayes, D. G., & Gulari, E. (1990). Esterification reactions of lipase in reverse micelles. Biotechnology and Bioengineering, 35, 793–801.
Guo, P., Zheng, C., Huang, F., Zheng, M., Deng, Q., & Li, W. (2013). Ultrasonic pretreatment for lipase-catalyzed synthesis of 4-methoxy cinnamoyl glycerol. Journal of Molecular Catalysis B: Enzymatic, 93, 73–78.
Fiametti, K. G., Sychoski, M. M., Cesaro, A. D., Furigo, A., Bretanha, L. C., Pereira, C. M. P., Treichel, H., Oliveira, D., & Oliveira, J. V. (2011). Ultrasound irradiation promoted efficient solvent-free lipase-catalyzed production of mono- and di-acylglycerols from olive oil. Ultrasonics Sonochemistry, 18, 981–987.
Zheng, M. M., Wang, L., Huang, F. H., Dong, L., Guo, P. M., Deng, Q. C., Li, W. L., & Zheng, C. (2012). Ultrasonic pretreatment for lipase-catalyzed synthesis of phytosterol esters with different acyl donors. Ultrasonics Sonochemistry, 19, 1015–1020.
Shah, S., & Gupta, M. N. (2008). The effect of ultrasonic pre-treatment on the catalytic activity of lipases in aqueous and non-aqueous media. Chemistry Central Journal, 2, 1–9.
Talukder, M. M. R., Zaman, M. M., Hayashi, Y., Wu, J. C., & Kawanishi, T. (2006). Ultrasonication enhanced hydrolytic activity of lipase in water/isooctane two-phase systems. Biocatal. Biotransfor., 24, 189–194.
Krishnamurthy, R., Lumpkin, J. A., & Sridhar, R. (2000). Inactivation of lysozyme by sonication under conditions relevant to microencapsulation. International Journal of Pharmaceutics, 205, 23–34.
Liu, Y., Jin, Q., Shan, L., Liu, Y., Shen, W., & Wang, X. (2008). The effect of ultrasound on lipase-catalyzed hydrolysis of soy oil in solvent-free system. Ultrasonics Sonochemistry, 15, 402–407.
Yu, Z. L., Zeng, W. C., Zhang, W. H., Liao, X. P., & Shi, B. (2014). Effect of ultrasound on the activity and conformation of a-amylase, papain and pepsin. Ultrasonics Sonochemistry, 21, 930–936.
Basto, C., Silva, C. J., Gu-bitz, G., & Cavaco-Paulo, A. (2007). Stability and decolourization ability of trametes villosa laccase in liquid ultrasonic fields. Ultrasonics Sonochemistry, 14, 355–362.
Tian, Z. M., Wan, M. X., Wang, S. P., & Kang, J. (2004). Effects of ultrasound and additives on the function and structure of trypsin. Ultrasonics Sonochemistry, 11, 399–404.
Lowry, R. R., & Tinsely, I. J. (1979). Rapid colorimetric determination of free fatty acids. Journal of the American Oil Chemists’ Society, 53, 470–472.
Wang, J., Wang, S., Li, Z., Gu, S., Wu, X., & Wu, F. (2015). Ultrasound irradiation accelerates the lipase-catalyzed synthesis of methyl caffeate in an ionic liquid. Journal of Molecular Catalysis B: Enzymatic, 111, 21–28.
Sakakibara, M., Wang, D., Takahashi, R., Takahashi, K., & Mori, S. (1996). Influence of ultrasound irradiation on hydrolysis of sucrose catalyzed by invertase. Enzyme Microbial Technology, 18, 444–448.
Bansode, S. R., & Rathod, V. K. (2014). Ultrasound assisted lipase catalyzed synthesis of isoamyl butyrate. Process Biochemistry, 49, 1297–1303.
Mills, A., & Holland, C. (1995). Effect of ultrasound on the kinetics of oxidation of octan-2-ol and other secondary alcohols with sodium bromate, mediated by ruthenium tetraoxide in a biphasic system. Ultrasonics Sonochemistry, 2, 33–38.
Sinisterra, J. V. (1992). Application of ultrasound to biotechnology: an overview. Ultrasonics, 30, 280–285.
Lerin, L. A., Loss, R. A., Remonatto, D., Zenevicz, M. C., Balen, M., Netto, V. O., Ninow, J. L., Trentin, C. M., Oliveira, J. V., & Oliveira, D. (2014). A review on lipase-catalyzed reactions in ultrasound-assisted systems. Bioprocess and Biosystems Engineering, 37, 2381–2394.
Bracey, E., Stenning, R. A., & Brooker, B. E. (1998). Relating the microstructure of enzyme dispersions in organic solvents to their kinetic behavior. Enzyme Microbial Technology, 22, 147–151.
Azevedo, A. M., Fonseca, L. P., & Prazeres, D. M. (1999). Stability and stabilization of penicillin acylase. Journal of Chemical Technology & Biotechnology, 74, 1110–1116.
Rawat, S., Suri, C. R., & Sahoo, D. K. (2010). Molecular mechanism of polyethylene glycol mediated stabilization of protein. Biochemical and Biophysical Research Communications, 392, 561–566.
Wu, J., Wang, Z., Lin, W., & Chen, S. (2013). Investigation of the interaction between poly (ethylene glycol) and protein molecules using low field nuclear magnetic resonance. Acta Biomaterialia, 9, 6414–6420.
Farruggia, B., Nerli, B., Nuci, H. D., Rigatusso, R., & Pico, G. (1999). Thermal features of the bovine serum albumin unfolding by polyethylene glycols. International Journal of Biological Macromolecules, 26, 23–33.
Verceta, A., Burgosa, J., Crelier, S., & Lopez-Buesa, P. (2001). Inactivation of proteases and lipases by ultrasound. Innovative Food Science & Emerging Technologies, 2, 139–150.
Li, C., Li, W., Holler, T. P., Gu, Z., & Li, Z. (2014). Polyethylene glycols enhance the thermostability of β-cyclodextrin glycosyltransferase from Bacillus circulans. Food Chemistry, 164, 17–22.
Klibanov, A. M. (1983). Stabilization of enzyme against thermal inactivation. Advances in Applied Microbiology, 29, 1–28.
Azizi, A., Ranjbar, B., Khajeh, K., Ghodselahi, T., Hoornam, S., Mobasheri, H., & Ganjalikhany, M. R. (2011). Effects of trehalose and sorbitol on the activity and structure of Pseudomonas cepacia lipase: spectroscopic insight. International Journal of Biological Macromolecules, 49, 652–656.
Talukder, M. M. R., Takeyama, T., Hayashi, Y., Wu, J. C., Kawanishi, T., Shimizu, Y., & Ogino, C. (2003). Improvement in enzyme activity and stability by addition of low molecular weight polyethylene glycol to sodium bis(2-ethyl-L-hexyl)sulfosuccinate/isooctane reverse micellar system. Applied Biochemistry and Biotechnology, 110, 101–111.
Talukder, M. M. R., Zaman, M. M., Hayashi, Y., Wu, J. C., & Kawanishi, T. (2007). Thermostability of Chromobacterium viscosum lipase in AOT/isooctane reverse micelle. Applied Biochemistry and Biotechnology, 141, 77–83.
Ranjbar, B., & Gill, P. (2009). Circular dichroism techniques: biomolecular and nanostructural analyses. Chemical Biology & Drug Design, 74, 101–120.
Bashari, M., Eibaid, A., Wanga, J., Tian, Y., Xu, X., & Jin, Z. (2013). Influence of low ultrasound intensity on the degradation of dextran catalyzed by dextranase. Ultrasonics Sonochemistry, 20, 155–161.
Huang, N., Chenga, X., Hu, W., & Pan, S. (2015). Inactivation, aggregation, secondary and tertiary structural changes of germin-like protein in Satsuma mandarine with high polyphenol oxidase activity induced by ultrasonic processing. Biophysical Chemistry, 197, 18–24.
Kelly, S. M., & Price, N. C. (1997). The application of circular dichroism to studies of protein folding and unfolding. Biochimica et Biophysica Acta, 1338, 161–185.
Kang, Y., Marangoni, A. G., & Yada, R. Y. (1994). Effect of two polar organic-aqueous solvent systems on the structure of proteases III, papain and trypsin. Journal of Food Biochemistry, 17, 389–394.
Melo, E. P., Taipa, M. A., Castellar, M. R., Costa, S. M. B., & Cabral, J. M. S. (2000). A spectroscopic analysis of thermal stability of the Chromobacterium viscosum lipase. Biophysical Chemistry, 87, 111–120.
Samanta, N., Mahanta, D. D., Hazra, S., Kumar, G. S., & Mitra, R. K. (2014). Short chain polyethylene glycols unusually assist thermal unfolding of human serum albumin. Biochimie, 104, 81–89.
De Cordt, S., Hendrickx, M., Maesmans, G., & Tobback, P. (1994). The influence of polyalcohols and carbohydrates on the thermostability of α-amylase. Biotechnology and Bioengineering, 43, 107–114.
Acknowledgments
Financial support from the Agency for Science Technology and Research (A*STAR) of Singapore is gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Talukder, M...M.R., Shiong, S.C.S. Stabilization of Chromobacterium viscosum Lipase (CVL) Against Ultrasound Inactivation by the Pretreatment with Polyethylene Glycol (PEG). Appl Biochem Biotechnol 177, 1742–1752 (2015). https://doi.org/10.1007/s12010-015-1850-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12010-015-1850-5