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Cloning, Expression, and Characterization of a Thermotolerant β-agarase from Simiduia sp. SH-4

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Abstract

The gene coding for a thermotolerant β-agarase from an isolated Simiduia sp. SH-4 was cloned, recombinantly expressed, and characterized after purification. This gene was sequenced after cassette mediated polymerase chain reaction and composed of an open reading frame of 1,809 base pairs, encoding a protein of 66.2 kilodaltons comprising of 602 amino acid residues. The amino acids sequence showed 74% homology with β-agarase of Simiduia agarivorans. A new β-agarase gene corresponding to mature protein of 577 amino acids was recombinantly expressed and purified by chitin bead column to homogeneity. The maximal specific activity was 505.07 U/mg at 50oC in Tris/HCl (pH 6.0) buffer. Recombinant β-agarase hydrolyzed agar into neoagarotetraose (57%) and neoagarohexaose (43%). It generated products from melted and non-melted powder agar and agarose at 30-50oC, meaning cheap agar materials could be used with energy- and costsavings. Thus, recombinant β-agarase could be used for industrial production of neoagarotetraose and neoagarohexaose.

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Authors and Affiliations

  1. Department of Green-Chemistry Convergence Engineering, Graduate School, Silla University, Busan, 617736, Korea

    Jae-Deog Kim & Sang-Hyeon Lee

  2. Department of Bioscience, Graduate School, Silla University, Busan, 617736, Korea

    Dong-Geun Lee & Sang-Hyeon Lee

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  1. Jae-Deog Kim
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  2. Dong-Geun Lee
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  3. Sang-Hyeon Lee
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Correspondence to Sang-Hyeon Lee.

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Kim, JD., Lee, DG. & Lee, SH. Cloning, Expression, and Characterization of a Thermotolerant β-agarase from Simiduia sp. SH-4. Biotechnol Bioproc E 23, 525–531 (2018). https://doi.org/10.1007/s12257-018-0072-4

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  • DOI: https://doi.org/10.1007/s12257-018-0072-4

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