FOOD SCIENCE ›› 2013, Vol. 34 ›› Issue (21): 217-220.doi: 10.7506/spkx1002-6630-201321044

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Screening of Enterobacter sakazakii-binding Peptides by Phage Display Technique

QU Wei,YUAN Jing-jing,ZHU Wei-dong,WANG Ming-ming,LIU Jian*   

  1. School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei 230009, China
  • Received:2012-11-27 Revised:2013-09-25 Online:2013-11-15 Published:2013-10-28
  • Contact: Jian LIU E-mail:liujian509@hfut.edu.cn

Abstract:

In this study, we used a phage-displayed random peptide library to identify phage clones, and the displayed
peptides could specifically and strongly bind to the cell surface of Enterobacter sakazakii. The capability of the phage clones
to interact specifically with Enterobacter sakazakii was demonstrated by using enzyme-linked immunosorbent assay (ELISA).
We assessed the selectivity of phage-bacteria binding by comparing the binding capability of the selected clones to the target
bacteria and a panel of other bacterial species. Tween-20 concentration, and binding and elution time were optimized to
enhance the stringency of selection or elution and to obtain a consensus binding sequence. After four rounds of screening,
positive phage clone DNAs were sequenced and their specific binding activity was identified by ELISA. The affinity of
the phage clones encoding QNDGTPR peptides was significantly higher than that from the control group. No phage clone
significantly bound to other selected bacterial species. Overall, Enterobacter sakazakii-binding peptide QNDGTPR was
screened and obtained by a random peptide library and the peptide may be used to develop novel diagnostic probes.

Key words: Enterobacter sakazakii, phage display, specific peptides

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