FOOD SCIENCE ›› 2018, Vol. 39 ›› Issue (6): 69-74.doi: 10.7506/spkx1002-6630-201806012

• Bioengineering • Previous Articles     Next Articles

Cloning, Expression and Purification of the Nitrite Reductase Gene from Bacillus cereus LJ01

CHEN Simin, LUO Tonghui, FEI Yongtao, WU Jianfeng, LIU Dongmei*   

  1. (School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China)
  • Online:2018-03-25 Published:2018-03-14

Abstract: A nitrite reductase (NiR) gene was cloned from Bacillus cereus LJ01 and expressed in Escherichia coli. The recombinant enzyme was purified by His-Tag nickel affinity chromatography (Ni Sepharose 6 Fast Flow) and DEAE Sepharose Fast Flow ion exchange chromatography for characterization. The results showed that the recombinant NiR gene contained a 1 623 bp-length ORF encoding 540 amino acids, and the molecular weight was about 67 ku. Iron and copper ions were simultaneously present in the enzyme and their contents were 51.0 and 184.5 mg/kg, respectively. The results of circular dichroism showed that the helix structure accounted for the largest proportion of the recombinant NiR.

Key words: nitrite reductase, Bacillus cereus, gene cloning, protein expression, engineered strain

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