Citation: GUO Jing, YAN Jiaze, GUO Ming, JIN Yan. Application of reversed-phase liquid chromatography-tandem mass spectrometry in the identification of protein and bioactivity peptides from rape bee pollen[J]. Chinese Journal of Chromatography, ;2014, 32(3): 284-289. doi: 10.3724/SP.J.1123.2013.12032 shu

Application of reversed-phase liquid chromatography-tandem mass spectrometry in the identification of protein and bioactivity peptides from rape bee pollen

GUO Jing ^1,2 ,
YAN Jiaze ² ,
GUO Ming ^1,, ,
JIN Yan ^2,,

1.
1. College of Environmental and Chemical Engineering, Dalian University, Dalian 116622, China;

Corresponding author: GUO Ming, JIN Yan,
Received Date: 27 December 2013
Available Online: 13 January 2014
Fund Project: 东北特色生物资源综合利用项目(2013SYHZ0020). (2013SYHZ0020)

Based on the shotgun proteomic method, rape bee pollen protein was prepared with ultrasonic extraction and digested by trypsin, then separated and sequenced by reversed-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS), followed by protein database searching. After the above analysis, 353 peptides were identified and the molecular biological functions of 239 proteins have been known. The identified molecular biological functions of these proteins mainly included binding activity, enzyme activity, transporter activity, inhibitor activity and so on. Five peptides were obtained after the screening and appropriate amino acid modification among the identified 353 peptides, according to the relationship between the sequence structure of the bioactivity peptide and angiotensin converting enzyme (ACE) inhibitor activity. The five peptides were speculated to have ACE inhibitor activity and synthesized to detect ACE inhibitor activity. The results showed that all of the five peptides had good ACE inhibitor activity. The peptides of AELDIVLALF and LAVNLIPFP among the five peptides displayed especially good ACE inhibition with half maximal inhibitory concentration (IC₅₀) of (10.65±0.50) μmol/L and (23.66±1.08) μmol/L, respectively. This method is rapid, low-cost and achieves the goal of high-throughput screening of bioactivity peptides that greatly shorten the period of identification compared with traditional methods.
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