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Molten Globule-Triggered Inactivation of a Thermostable and Solvent Stable Lipase in Hydrophilic Solvents

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Abstract

The use of lipase in hydrophilic solvent is usually hampered by inactivation. The solvent stability of a recombinant solvent stable lipase isolated from thermostable Bacillus sp. strain 42 (Lip 42), in DMSO and methanol were studied at different solvent-water compositions. The enzymatic activities were retained in up to 45% v/v solvent compositions. The near-UV CD spectra indicated that tertiary structures were perturbed at 60% v/v and above. Far-UV CD in methanol indicated the secondary structure in Lip 42 was retained throughout all solvent compositions. Fluorescence studies indicated formations of molten globules in solvent compositions of 60% v/v and above. The enzyme was able to retain its secondary structures in the presence of methanol; however, there was a general reduction in β-sheet and an increase in α-helix contents. The H-bonding arrangements triggered in methanol and DMSO, respectively, caused different forms of tertiary structure perturbations on Lip 42, despite both showing partial denaturation with molten globule formations.

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Abbreviations

ANS:

8-Anilino-1-napthalene sulphonic acid

DMSO:

Dimethyl sulfoxide

UV CD:

Ultra violet circular dichroism

MG:

Molten globule

Phe:

Residue phenylalanine

Trp:

Residue tryptophan

Tyr:

Residue tyrosine

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Acknowledgments

This project was funded by the Ministry of Higher Education Malaysia (MOHE) under The Fundamental Research Grant Scheme (FRGS) (project number 01-01-07-009FR) and Tengku Haziyamin Tengku Abdul Hamid was supported by International Islamic University Malaysia for personal allowances.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Tengku Haziyamin Tengku Abdul Hamid & Raja Noor Zaliha Raja Abd Rahman

  2. Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, University Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Abu Bakar Salleh

  3. Department of Chemistry, Faculty of Science, University Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Mahiran Basri

Authors
  1. Tengku Haziyamin Tengku Abdul Hamid
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  2. Raja Noor Zaliha Raja Abd Rahman
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  3. Abu Bakar Salleh
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  4. Mahiran Basri
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Correspondence to Raja Noor Zaliha Raja Abd Rahman.

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Hamid, T.H.T.A., Rahman, R.N.Z.R.A., Salleh, A.B. et al. Molten Globule-Triggered Inactivation of a Thermostable and Solvent Stable Lipase in Hydrophilic Solvents. Protein J 29, 290–297 (2010). https://doi.org/10.1007/s10930-010-9251-7

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  • DOI: https://doi.org/10.1007/s10930-010-9251-7

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