Food and Fermentation Industries >

2019 , Vol. 45 >Issue 3: 77 - 82

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.018288

Cloning and expression of the alginate lyase gene B1SM frommarine Pseudoalteromonas sp.

  • B1WU Chenshuo ,
  • LI Xiaoyue ,
  • QIN Mingzhen ,
  • YAN Fen
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  • (College of Biological Science and Technology, Fuzhou University, Fuzhou 350108,China)

Received date: 2018-07-16

  Online published: 2019-03-11

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Abstract

The recombinant alginate lyase B1SM was isolated and purified, and its enzymatic properties were characterized. The alginate lyase gene B1SM was amplified from the genome of Pseudomonas aeruginosa B1 by Touch down polymerase chain reaction (PCR) and Thermal Asymmetric Interlaced PCR (TAIL-PCR). The gene B1SM was inserted into a pGEX-4T-1 expression vector and expressed in Escherichia coli BL21 (DE3). The results showed that the optimal pH and temperature for B1SM were 8.0 and 25℃, respectively. Moreover, B1SM maintained more than 60% of its relative enzyme activity after incubating at 25℃ for 1 h at pH=7.0-9.0, indicating that it had a good pH stability. Furthermore, it maintained its relative enzyme activity to more than 60% after incubating at 30 ℃ and at pH=8.0 for 1 h. It was concluded that heterologous expression of alginate lyase gene B1SM was achievable in E. coli BL21 (DE3), which laid a foundation for preparing and further study of alginate lyase.

Key words： alginate lyase; Pseudoalteromonas sp.; polymerase chain reaction (PCR)

Cite this article

B1WU Chenshuo , LI Xiaoyue , QIN Mingzhen , YAN Fen . Cloning and expression of the alginate lyase gene B1SM frommarine Pseudoalteromonas sp.[J]. Food and Fermentation Industries, 2019 , 45(3) : 77 -82 . DOI: 10.13995/j.cnki.11-1802/ts.018288

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