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Biochemical and Molecular Characterizations of a Novel pH- and Temperature-Stable Pectate Lyase from Bacillus amyloliquefaciens S6 for Industrial Application

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Abstract

In this paper, we report cloning of a pectate lyase gene from Bacillus amyloliquefaciens S6 (pelS6), and biochemical characterization of the recombinant pectate lyase. PelS6 was found to be identical with B. subtilis 168 pel enzyme with 100% amino acid sequence homology. Although these two are genetically very close, they are distinctly different in physiology. pelS6 gene encodes a 421-aa protein with a molecular mass of 65,75 kDa. Enzyme activity increased from 12.8 ± 0.3 to 49.6 ± 0.4 units/mg after cloning. The relative enzyme activity of the recPel S6 ranged from 80% to 100% at pH between 4 and 14. It was quite stable at different temperature values ranging from 15 to 90 °C. The recPEL S6 showed a maximal activity at pH 10 and at 60 °C. 0.5 mM of CaCl2 is the most effective metal ion on the recPEL S6 as demonstrated by its increased relative activity with 473%. recPEL S6 remained stable at − 20 °C for 18 months. In addition recPEL S6 increased juice clarity. This study introduces a novel bacterial pectate lyase enzyme with its characteristic capability of being highly thermostable, thermotolerant, and active over a wide range of pH, meaning that it can work at both acidic and alkaline environments, which are the most preferred properties in the industry.

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Acknowledgements

This research was supported by the BAP (Project No. 05/2016-25).

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Authors and Affiliations

  1. Department of Biology, Gazi University, Ankara, Turkey

    Seda Bekli & Belma Aslim

  2. Department of Molecular Biology and Genetics, Burdur Mehmet Akif Ersoy University, Burdur, Turkey

    Busra Aktas

  3. Department of Biology, Karadeniz Technical University, Trabzon, Turkey

    Donus Gencer

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  1. Seda Bekli
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Bekli, S., Aktas, B., Gencer, D. et al. Biochemical and Molecular Characterizations of a Novel pH- and Temperature-Stable Pectate Lyase from Bacillus amyloliquefaciens S6 for Industrial Application. Mol Biotechnol 61, 681–693 (2019). https://doi.org/10.1007/s12033-019-00194-2

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