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 express...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-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.展开更多
A novel lipase gene(lip4346)encoding a primary translation product with 176 amino acids was screened from the genome fine mapping of the macroalgae-associated bacterial strain Microbulbifer sp.YNDZ01.Macroalgae were c...A novel lipase gene(lip4346)encoding a primary translation product with 176 amino acids was screened from the genome fine mapping of the macroalgae-associated bacterial strain Microbulbifer sp.YNDZ01.Macroalgae were collected from the coast of the Halmahera Island of Indonesia.The lip4346 gene was cloned and heterologously expressed in Escherichia coli.The purified recombinant Lip4346 protein had a molecular mass of 19 k Da,a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),and a maximum enzyme activity of 31.2 U m L-1.The optimal temperature and pH for the lipase activity of Lip4346 were 70℃and 10.0,respectively.Lip4346 was tolerant with a number of organic solvents and detergents,and was active toward triacylglycerols and p-nitrophenyl esters with short-and medium-chain lengths.The unique characteristics of Lip4346 indicate that it is a promising nonaqueous biocatalyst for industrial applications.展开更多
基金The Natural Science Foundation of Fujian Province of China under contract No.2016J01162the Program for New Century Excellent Talents in Fujian Province University,China under contract No.B15139
文摘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-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.
基金financially supported by the China Ocean Mineral Resources R&D Association project(No.DY 135-B2-11)the China-ASEAN Maritime Cooperation Fund。
文摘A novel lipase gene(lip4346)encoding a primary translation product with 176 amino acids was screened from the genome fine mapping of the macroalgae-associated bacterial strain Microbulbifer sp.YNDZ01.Macroalgae were collected from the coast of the Halmahera Island of Indonesia.The lip4346 gene was cloned and heterologously expressed in Escherichia coli.The purified recombinant Lip4346 protein had a molecular mass of 19 k Da,a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE),and a maximum enzyme activity of 31.2 U m L-1.The optimal temperature and pH for the lipase activity of Lip4346 were 70℃and 10.0,respectively.Lip4346 was tolerant with a number of organic solvents and detergents,and was active toward triacylglycerols and p-nitrophenyl esters with short-and medium-chain lengths.The unique characteristics of Lip4346 indicate that it is a promising nonaqueous biocatalyst for industrial applications.
