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Expression, purification, and characterization of a recombined fibrinolytic enzyme from endophytic Paenibacillus polymyxa EJS-3 in Escherichia coli

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Abstract

The gene encoding the fibrinolytic enzyme (PPFE-I) from Paenibacillus polymyxa EJS-3 was cloned and sequenced (Genbank No. KC176802). The 1773 bp gene encoded 590 amino acids and had low homology with other known fibrinolytic enzymes. PPFE-I was soluble and expressed in E. coli BL21 to enhance the activity. The recombinant enzyme (rPPFE-I) was purified to homogeneity, and enzymatic properties were characterized. The optimal temperature and pH for rPPFE-I were 37°C and 7.5, respectively. Observed activities of rPPFE-I were highest in the presence of Zn2+, Mg2+, and Fe2+ and the lowest in the presence of Ca2+ and Cu2+. The rPPFE-I activity was strongly inhibited by PMSF. The α-chain was affected by the rPPFE-I cleavage speed of fibrinogen, followed by the b and γ chains. The inhibitory effects of rPPFE-I against ADP-induced human platelet aggregation were dose-dependent with an IC50 value of 1.54 μM.

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Authors and Affiliations

  1. College of Food Science and Technology, Nanjing Agricultural University, Weigang, Nanjing, 210095, China

    Fengxia Lv, Chong Zhang, Fangfang Guo, Xiaomei Bie, Hui Qian & Zhaoxin Lu

  2. Department of Nutrition and Food Science, University of Maryland, College Park, Maryland, 20740, USA

    Yingjian Lu

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  1. Fengxia Lv
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  2. Chong Zhang
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  3. Fangfang Guo
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  4. Yingjian Lu
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  7. Zhaoxin Lu
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Correspondence to Zhaoxin Lu.

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Lv, F., Zhang, C., Guo, F. et al. Expression, purification, and characterization of a recombined fibrinolytic enzyme from endophytic Paenibacillus polymyxa EJS-3 in Escherichia coli . Food Sci Biotechnol 24, 125–131 (2015). https://doi.org/10.1007/s10068-015-0018-y

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  • DOI: https://doi.org/10.1007/s10068-015-0018-y

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