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Protein Engineering of Bacillus thermocatenulatus Lipase via Deletion of the α5 Helix

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Abstract

Lipases from Bacillus thermocatenulatus are a member of superfamily of α/β hydrolase, but there are structural differences between them. In this work, we focused on the α5 helix of B. thermocatenulatus lipase (BTL2) which is absent in canonical α/β hydrolase fold. In silico study showed that the α5 helix is a region that causes disorder in BTL2 protein. So, the α5 helix (residues 131 to 150) has been deleted from the B. thermocatenulatus lipase gene (BTL2) and the remain (Δα5-BTL2) has been expressed in Pichia pastoris yeast. The α5 deletion results in increase of enzyme-specific activity in the presence of tributyrin, tricaproin, tricaprylin, tricaprin, trilaurin, and olive oil (C18) substrates by 1.4-, 1.7-, 2.0-, 1.2-, 1.75-, and 1.95-fold, respectively. Also, deletion leads to increase in enzyme activity in different temperatures and pHs, whereas it did not significantly affect on enzyme activity in the presence of organic solvents, metal ions, and detergents.

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Abbreviations

BMMH:

Buffered minimal medium containing histidine

YPG:

Yeast extract peptone dextrose medium

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Acknowledgments

We would like to express our appreciation and thanks to Mrs. F. Rahimi and Dr. P. Farrokh for their help on the accomplishment of this study. We are also grateful to Mr. H. Jamali for the language editing of our manuscript.

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Authors and Affiliations

  1. National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, Iran

    Ali Asghar Karkhane, Fatemeh Tabandeh, Saeed Aminzadeh, Bagher Yakhchali & Mehdi Shamsara

  2. Department of Biotechnology, Isfahan University of Technology, Isfahan, 84156-8311, Iran

    Nastaran Goodarzi & Aghafakhr Mirlohi

  3. Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran

    Ibrahim Torktas

  4. Department of Plant Breeding, University of Zanjan, Zanjan, Iran

    Mozhdeh Al-Sadat Ghafouri

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  1. Nastaran Goodarzi
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  2. Ali Asghar Karkhane
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  3. Aghafakhr Mirlohi
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Correspondence to Ali Asghar Karkhane.

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Goodarzi, N., Karkhane, A.A., Mirlohi, A. et al. Protein Engineering of Bacillus thermocatenulatus Lipase via Deletion of the α5 Helix. Appl Biochem Biotechnol 174, 339–351 (2014). https://doi.org/10.1007/s12010-014-1063-3

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  • DOI: https://doi.org/10.1007/s12010-014-1063-3

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