Cloning, Expression, and Characterization of a Thermotolerant β-agarase from Simiduia sp. SH-4
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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.
Keywordsβ-agarase cloning expression purification Simiduia sp.
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- 2.Kobayashi, R., M. Takisada, T. Suzuki, K. Kirimura, and S. Usami (1997) Neoagarobiose as a novel moisturizer with whitening effect. Biosci. Biotechnol. Biochem. 61: 62–63.Google Scholar
- 8.Lee, D. G., M. K. Jang, O. H. Lee, N. Y. Kim, S. A. Ju, and S. H. Lee (2008) Over-production of a glycoside hydrolase family 50 ß-agarase from Agarivorans sp. JA-1 in Bacillus subtilis and the whitening effect of its product. Biotechnol. Lett. 30: 911–918.Google Scholar
- 12.Tawara, M., A. Sakatoku, R. E. Tiodjio, D. Tanaka, and S. Nakamura (2015) Cloning and characterization of a novel agarase from a newly isolated bacterium Simiduia sp. strain TM-2 able to degrade various seaweeds. Appl. Biochem. Biotechnol. 177: 610–623.Google Scholar
- 15.Lin, S. Y., W. Y. Shieh, J. S. Chen, and S. L. Tang (2013) Complete genome sequence of Simiduia agarivorans SA1T, a marine bacterium able to degrade a variety of polysaccharides. Genome Announc. 1: e00039–12.Google Scholar