A nitrilase gene cyc705 from Arthrobacter aures-

نویسندگان

  • Wenwen Cai
  • Erzheng Su
  • Shujing Zhu
  • Yuhong Ren
  • Dongzhi Wei
چکیده

cens CYC705 for synthesis of iminodiacetic acid (IDA) was cloned. This gene contained a 930 bp ORF, which encoded a polypeptide of 310 amino acids. A recombinant Escherichia coli BL21(DE3)/ pET28a-cyc705 was constructed to achieve the heterologous expression of cyc705. This recombinant nitrilase was purified to homogeneity with a molecular weight of 36.7 kDa on SDS-PAGE and mass spectrometry, and characterized to be an oligomer of 14 subunits by gel permeation chromatography. Using iminodiacetonitrile (IDAN) as the substrate, the Vmax, Km, kcat and kcat / Km were 9.05 U mg-1, 43.17 mM−1, 94.1 min−1 and 2.18×103 min−1 M−1, respectively. The optimum temperature and pH were 25°C and 5.8. The suitable substrates for the purified nitrilase were short-chain aliphatic dinitriles. High concentration of IDAN could be hydrolyzed to IDA in a shorter time.

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تاریخ انتشار 2014