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Improving Activity of Salt-Lyophilized Enzymes in Organic Media

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Abstract

Lyophilization with salts has been identified as an important method of activating enzymes in organic media. Using salt-activated enzymes to transform molecules tethered to solid surfaces in organic phase requires solubilization of enzymes in the solvents. Methods of improving performance of salt-lyophilized enzymes, further, via chemical modification, and use of surfactants and surfactants to create fine emulsions prior to lyophilization are investigated. The reaction system used is transesterification of N-acetyl phenylalanine ethyl ester with methanol or propanol. Initial rate of formation of amino acid esters by subtilisin Carlsberg (SC) was studied and found to increase two to sevenfold by either chemical modification or addition of surfactants in certain solvents, relative to the salt (only)-lyophilized enzyme. The method to prepare highly dispersed enzymes in a salt-surfactant milieu also improved activity by two to threefold. To test the effect of chemical modification on derivatization of drug molecules, acylation of bergenin was investigated using chemically modified SC.

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Acknowledgements

The authors wish to acknowledge support from the National Institute of Health under the Bioengineering Research Partnerships. Initial project direction and assistance from Dr. Jonathan Dordick is greatly appreciated.

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  1. Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, 37831-6226, USA

    Abhijeet P. Borole & Brian H. Davison

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  1. Abhijeet P. Borole
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  2. Brian H. Davison
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Correspondence to Abhijeet P. Borole.

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The submitted manuscript has been authored by a contractor of the US Government under contract No. DE-AC05-00OR22725. Accordingly, the US Government retains a nonexclusive, royalty-free license to publish or reproduce the published form of this contribution, or allow others to do so, for US Government purposes.

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Borole, A.P., Davison, B.H. Improving Activity of Salt-Lyophilized Enzymes in Organic Media. Appl Biochem Biotechnol 146, 215–222 (2008). https://doi.org/10.1007/s12010-007-8033-y

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  • DOI: https://doi.org/10.1007/s12010-007-8033-y

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