Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate <i>Microbulbifer</i> sp. AG1

ZHU Yanbing; GAO He; LI Hebin; NI Hui; JIANG Zedong; LI Lijun; XIAO Anfeng

doi:10.1007/s13131-019-1349-y

Volume 38 Issue 2

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Article Navigation > Acta Oceanologica Sinica > 2019 > 38(2): 96-106

ZHU Yanbing, GAO He, LI Hebin, NI Hui, JIANG Zedong, LI Lijun, XIAO Anfeng. Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate Microbulbifer sp. AG1[J]. Acta Oceanologica Sinica, 2019, 38(2): 96-106. doi: 10.1007/s13131-019-1349-y

Citation:

ZHU Yanbing, GAO He, LI Hebin, NI Hui, JIANG Zedong, LI Lijun, XIAO Anfeng. Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate Microbulbifer sp. AG1[J]. Acta Oceanologica Sinica, 2019, 38(2): 96-106. doi: 10.1007/s13131-019-1349-y

Citation:

ZHU Yanbing, GAO He, LI Hebin, NI Hui, JIANG Zedong, LI Lijun, XIAO Anfeng. Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate Microbulbifer sp. AG1[J]. Acta Oceanologica Sinica, 2019, 38(2): 96-106. doi: 10.1007/s13131-019-1349-y

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Overexpression and characterization of a thermostable β-agarase producing neoagarotetraose from a marine isolate Microbulbifer sp. AG1

doi: 10.1007/s13131-019-1349-y

1.
College of Food and Biological Engineering, Jimei University, Xiamen 361021, China;Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China;Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China;Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen 361021, China
2.
College of Food and Biological Engineering, Jimei University, Xiamen 361021, China
3.
Department of Pharmacy, Xiamen Medical College, Xiamen 361023, China

Received Date: 2017-01-14

Abstract

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℃ and 7.5, respectively. The recombinant agarase showed excellent thermostability with 67% and 19% of residual activities after incubation at 50℃ and 60℃ 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-D-galactopyranoside 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.
- thermostable,
- β-agarase,
- neoagarotetraose,
- Microbulbifer sp.

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References

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