Abstract
The transesterification-active enzyme bacillolysin was extracted into organic solvents such as isooctane by enzyme-AOT (bis (2-ethylhexyl) sulfosuccinate) ion-pairing preserving its natural second structure and catalytic activity. Extraction efficiency was affected by the interaction mode of the two phases, ionic strength, and pH of aqueous phase, surfactant and enzyme concentration. Magnetic stirring with phase mixing was favorable for the enzyme extraction. Optimal ionic strength and pH were 8 mM CaCl2 and 5.0, respectively. Critical number of AOT molecule for an enzyme molecule to be extracted into isooctane was 89. Optimal initial enzyme concentration in the aqueous phase was 7 mg mL−1 while the initial AOT concentration in isooctane was 3 mM. Within CMC (critical micellar concentration) of AOT in isooctane, the increase of initial AOT concentration enhanced the extraction efficiency.
Similar content being viewed by others
References
Altreuter, D. H., Dordick, J. S., & Clark, D. S. (2002). Optimization of ion-paired lipase for non-aqueous media: acylation of doxorubicin based on surface models of fatty acid esterification. Enzyme and Microbial Technology, 31, 10–19. DOI: 10.1016/S0141-0229(02)00092-3.
Bruno, F. F., Akkara, J. A., Ayyagari, M., Kaplan, D. L., Gross, R., Swift, G., & Dordick, J. S. (1995). Enzymic modification of insoluble amylose in organic solvents. Macromolecules, 28, 8881–8883. DOI: 10.1021/ma00130a028.
Castillo, B., Solá, R. J., Ferrer, A., Barletta, G., & Griebenow, K. (2007). Effect of PEG modification on subtilisin Carlsberg activity, enantioselectivity, and structural dynamics in 1,4-dioxane. Biotechnology and Bioengineering, 99, 9–17. DOI: 10.1002/bit.21510.
Gu, J. D., Fan, X. R., Wang, S. G., & Zhang, Y. (2009). Study of enzyme transesterification in solvents with fluorospectrophometry. Huaxue Tongbao, 72, 444–448.
He, P., Huang, J. L., Shi, Q. C., Wu, G. H., Chu, Z. C., & Huang, Z. L. (2009). Enzyme activity changes of papain and immobilized papain in organic solvents with different logP values. Journal of Northwest A&F University (Natural Science Edition), 37, 223–229.
Jurado, E., Camacho, F., Luzón, G., & Vicaria, J. M. (2004). Kinetic models of activity for β-galactosidases: influence of pH, ionic concentration and temperature. Enzyme and Microbial Technology, 34, 33–40. DOI: 10.1016/j.enzmictec.2003.07.004.
Konarzycka-Bessler, M., & Bornscheuer, U. T. (2003). A high-throughput-screening method for determining the synthetic activity of hydrolases. Angewandte Chemie International Edition, 42, 1418–1420. DOI: 10.1002/anie.200390365.
Paradkar, V. M., & Dordick, J. S. (1994). Mechanism of extraction of chymotrypsin into isooctane at very low concentrations of aerosol OT in the absence of reversed micelles. Biotechnology and Bioengineering, 43, 529–540. DOI: 10.1002/bit.260430614.
Pepić, I., Filipović-Grčić, J., & Jalšenjak, I. (2008). Interactions in a nonionic surfactant and chitosan mixtures. Colloids and Surfaces A, 327, 95–102. DOI: 10.1016/j.colsurfa.2008.06.009.
Persson, M., Mladenoska, I., Wehtje, E., & Adlercreutz, P. (2002). Preparation of lipases for use in organic solvents. Enzyme and Microbial Technology, 31, 833–841. DOI: 10.1016/S0141-0229(02)00184-9.
Rodakiewicz-Nowak, J., Monkiewicz, M., & Haber, J. (2002). Enzymatic activity of the A. bisporus tyrosinase in AOT/isooctane water-in-oil microemulsions. Colloids and Surfaces A, 208, 347–356. DOI: 10.1016/S0927-7757(02)00161-9.
Roy, I., & Gupta, M. N. (2005). Enhancing reaction rate for transesterification reaction catalyzed by Chromobacterium lipase. Enzyme and Microbial Technology, 36, 896–899. DOI: 10.1016/j.enzmictec.2005.01.022.
Roy, I., Sharma, A., & Gupta, M. N. (2004). Obtaining higher transesterification rates with subtilisin Carlsberg in nonaqueous media. Bioorganic & Medicinal Chemistry Letters, 14, 887–889. DOI: 10.1016/j.bmcl.2003.12.021.
Tonova, K., & Lazarova, Z. (2008). Reversed micelle solvents as tools of enzyme purification and enzyme-catalyzed conversion. Biotechnology Advances, 26, 516–532. DOI: 10.1016/j.biotechadv.2008.06.002.
Tsuru, D., McConn, J. D., & Yasunobu, K. T. (1965). Bacillus subtilis neutral proteinase II. Some physicochemical properties. Journal of Biological Chemistry, 240, 2415–2420.
Xie, J., & Hsieh, Y.-L. (2001). Enzyme-catalyzed transesterification of vinyl esters on cellulose solids. Journal of Polymer Science Part A: Polymer Chemistry, 39, 1931–1939. DOI: 10.1002/pola.1170.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhang, Y., Gu, J., Wang, S. et al. Preparation and properties of surfactant-bacillolysin ion-pair in organic solvents. Chem. Pap. 64, 443–449 (2010). https://doi.org/10.2478/s11696-010-0018-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2478/s11696-010-0018-6