Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (8): 196-201.

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Optimization Screening of Peptides Inhibition to H37Ra

Wu Congmei, Li Lingling, Guan Xiaoxia, Liu Xintao, Chen Ji, Yin Yuhe   

  1. School of Chemistry and Life Science, Changchun University of Technology, Changchun 130012
  • Revised:2014-03-31 Online:2014-08-15 Published:2014-08-01
  • Contact: 通讯作者: 殷玉和,男,讲师,研究方向:生物工程学;E-mail:yyhlxt72@sina.com
  • Supported by:
    吉林省科技发展计划项目(2008110)

Abstract: By inhibition experiments to determine the inhibitory effect of peptide compounds under different culture conditions, and measure theirs minimal inhibitory concentration(MIC). Adding Vc to compare inhibitory effect and future optimize screening H37Ra inhibitors. According to the results of ELISA test screen the phage peptide library, and then uses the Molecular docking software to simulate peptide docking with ICL protein crystal(1F8I). The successful docking peptide uses the solid-phase synthesis method of Fmoc for synthesis, and tests its biological activity. By preparative reversed phase HPLC purifies the synthesized peptides and detects mass spectrometry. Four peptides were screened, all of which have bacteriostatic effect, and correlated with the dose. The results showed that when the concentration between 800 and 1 500 μg/mL all group were bacteriostat. When the concentration is 500 μg/mL, only one peptide has bacteriostatic effect in the normal medium, while all cannot inhibit in the limit carbon medium. The inhibitory effect of No.2 peptide is best, and its MIC is 200 μg/mL in normal culture, while it is 500 μg/mL in carbon limitations. Positive control group, two media’s inhibitory effect were consistent, RIF’s and INH’s MIC were 0.8 μg/mL and 0.5 μg/mL, respectively. According to MIC results, the inhibitory effects of peptide, RIF and INH were improved when adding Vc to the normal culture, and to be a dose-dependent increase.

Key words: Peptides, MIC, Phage peptide library, Molecular docking, Solid-phase synthesis method of Fmoc