Abstract
An agarase gene containing 1 302 bp was cloned from Microbulbifer sp. AG1. It encoded a mature protein of 413 amino acids plus a 20-residue signal peptide. The recombinant enzyme without the signal peptide was expressed and purified from Escherichia coli BL21 (DE3). When agarose was used as a substrate, the optimal temperature and pH for the enzyme were 60°C and 7.5, respectively. The recombinant agarase showed excellent thermostability with 67% and 19% of residual activities after incubation at 50°C and 60°C for 1 h, respectively. Except SDS, the recombinant agarase had a relatively good resistance against the detected inhibitors, detergents and urea denaturant. Thin layer chromatography analysis and enzyme assay using p-nitrophenyl-α/β-Dgalactopyranoside revealed that the recombinant agarase was a β-agarase that degraded agarose into neoagarotetraose as the main end product. The enzymatic hydrolysis products with different degree of polymerization exhibited the antioxidant activities.
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Foundation item: The Natural Science Foundation of Fujian Province of China under contract No. 2016J01162; the Program for New Century Excellent Talents in Fujian Province University, China under contract No. B15139.
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Zhu, Y., Gao, H., Li, H. et al. Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate Microbulbifer sp. AG1. Acta Oceanol. Sin. 38, 96–106 (2019). https://doi.org/10.1007/s13131-019-1349-y
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DOI: https://doi.org/10.1007/s13131-019-1349-y