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Pentynyl dextran as a support matrix for immobilization of serine protease subtilisin Carlsberg and its use for transesterification of N-acetyl-l-phenylalanine ethyl ester in organic media

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Abstract

In this study, serine protease (subtilisin Carlsberg) was immobilized on pentynyl dextran (PyD, O–alkynyl ether of dextran, 1) and used for the transesterification of N-acetyl-l-phenylalanine ethyl ester (2) with different aliphatic (1-propanol, 1-butanol, 1-pentanol, 1-hexanol) and aromatic (benzyl alcohol, 2-phenyl ethanol, 4-phenyl-1-butanol) alcohols in tetrahydrofuran (THF). The effect of carbon chain length in aliphatic and aromatic alcohols on initial and average transesterification rate, transesterification activity of immobilized enzyme and yield of the reaction under selected reaction conditions was investigated. The transesterification reactivity of the enzyme and yield of the reaction increased as the chain length of the alcohols decreased. Furthermore, almost no change in yield was observed when the immobilized enzyme was repeatedly used for selected alcohols over six cycles. Intrinsic fluorescence analysis showed that the catalytic activity of the immobilized enzyme in THF was maintained due to retention of the tertiary structure of the enzyme after immobilization on PyD (1).

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Scheme 1
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Abbreviations

APAEE:

N-acetyl-l-phenylalanine ethyl ester (2)

DS:

Degree of substitution

EA:

Elemental analysis

PyD:

Pentynyl dextran (1)

SC:

Subtilisin Carlsberg

Trp:

Tryptophan

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Acknowledgments

Financial support of Konkuk University (KU Brain pool) is greatfully acknowledged. This work is also supported by the National Research Foundation of Korea Grant funded by the Korean Government (NRF-2011-0024008 and NRF-2011-619-E0002).

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Authors and Affiliations

  1. Department of Bioscience and Biotechnology, Center for Biotechnology Research in UBITA(CBRU), Konkuk University, Seoul, 143-701, Republic of Korea

    Muhammad Nazir Tahir, Eunae Cho & Seunho Jung

  2. Institute of Food Chemistry, Technische Universität Braunschweig, Schleinitzstr. 20, 38106, Braunschweig, Germany

    Petra Mischnick

  3. Department of Biological Science, Mokpo National University, Jeonnam, 534-729, Republic of Korea

    Jae Yung Lee

  4. Departments of Bacteriology and Genetics, and Molecular and Environmental Toxicology Center, University of Wisconsin, Madison, WI, USA

    Jae-Hyuk Yu

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  1. Muhammad Nazir Tahir
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  2. Eunae Cho
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Correspondence to Seunho Jung.

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Tahir, M.N., Cho, E., Mischnick, P. et al. Pentynyl dextran as a support matrix for immobilization of serine protease subtilisin Carlsberg and its use for transesterification of N-acetyl-l-phenylalanine ethyl ester in organic media. Bioprocess Biosyst Eng 37, 687–695 (2014). https://doi.org/10.1007/s00449-013-1038-8

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  • DOI: https://doi.org/10.1007/s00449-013-1038-8

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