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Sorbitol counteracts temperature- and chemical-induced denaturation of a recombinant α-amylase from alkaliphilic Bacillus sp. TS-23

  • Biocatalysis
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Enzymes are highly complex systems with a substantial degree of structural variability in their folded state. In the presence of cosolvents, fluctuations among vast numbers of folded and unfolded conformations occur via many different pathways; alternatively, certain conformations can be stabilized or destabilized. To understand the contribution of osmolytes to the stabilization of structural changes and enzymatic activity of a truncated Bacillus sp. TS-23 α-amylase (BACΔNC), we monitored amylolytic activity, circular dichroism, and fluorescence as a function of osmolytes. In the presence of trimethylamine N-oxide (TMAO) and sorbitol, BACΔNC activity was retained significantly at elevated temperatures. As compared to the control, the secondary structures of this enzyme were essentially conserved upon the addition of these two kinds of osmolytes. Fluorescence results revealed that the temperature-induced conformational change of BACΔNC was prevented by TMAO and sorbitol. However, glycerol did not provide profound protection against thermal denaturation of the enzyme. Sorbitol was further found to counteract guanidine hydrochloride- and SDS-induced denaturation of BACΔNC. Thus, some well-known naturally occurring osmolytes make a dominant contribution to the stabilization of BACΔNC.

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Acknowledgments

This work was supported by a research grant (NSC 97-2628-B-415-001-MY3) from National Science Council of Taiwan.

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Authors and Affiliations

  1. Department of Applied Chemistry, National Chiayi University, 300 Syuefu Road, Chiayi County, 60004, Taiwan

    Meng-Chun Chi, Tai-Jung Wu & Long-Liu Lin

  2. Department of Food Science and Technology, Hungkuang University, Shalu, Taichung County, 433, Taiwan

    Hsing-Ling Chen & Huei-Fen Lo

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  1. Meng-Chun Chi
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  2. Tai-Jung Wu
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Correspondence to Huei-Fen Lo or Long-Liu Lin.

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Chi, MC., Wu, TJ., Chen, HL. et al. Sorbitol counteracts temperature- and chemical-induced denaturation of a recombinant α-amylase from alkaliphilic Bacillus sp. TS-23. J Ind Microbiol Biotechnol 39, 1779–1788 (2012). https://doi.org/10.1007/s10295-012-1183-0

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