Discovery of the bioactive peptides secreted by Bifidobacterium using integrated MCX coupled with LC-MS and feature-based molecular networking

Shengshuang Chen; Guoxin Huang; Weilin Liao; Shilin Gong; Jianbo Xiao; Jiao Bai; W L Wendy Hsiao; Na Li; Jian-Lin Wu

doi:10.1016/j.foodchem.2021.129008

Discovery of the bioactive peptides secreted by Bifidobacterium using integrated MCX coupled with LC-MS and feature-based molecular networking

Food Chem. 2021 Jun 15:347:129008. doi: 10.1016/j.foodchem.2021.129008. Epub 2021 Jan 7.

Authors

Shengshuang Chen¹, Guoxin Huang¹, Weilin Liao¹, Shilin Gong¹, Jianbo Xiao², Jiao Bai³, W L Wendy Hsiao¹, Na Li⁴, Jian-Lin Wu⁵

Affiliations

¹ Macau Institute for Applied Research in Medicine and Health, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau.
² Institute of Chinese Medical Sciences, State Key Laboratory of Quality Research in Chinese Medicine, University of Macau, Macau.
³ Key Laboratory of Structure-Based Drug Design and Discovery, Ministry of Education, School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China.
⁴ Macau Institute for Applied Research in Medicine and Health, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau. Electronic address: nli@must.edu.mo.
⁵ Macau Institute for Applied Research in Medicine and Health, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau. Electronic address: jlwu@must.edu.mo.

PMID: 33484958
DOI: 10.1016/j.foodchem.2021.129008

Abstract

Probiotics can release many bioactive peptides that confer a myriad of benefits to the host health. However, exploring new bioactive peptides secreted by probiotics is hampered by lots of matrix-related interference peptides from the medium, and the low abundance. To this end, a new approach integrating mixed-mode cationic exchange based solid-phase extraction (MCX-SPE) coupled with liquid chromatography-mass spectrometry (LC-MS) and feature-based molecular networking (FBMN) was developed. FBMN's intuitive visualization results enabled twenty-five novel peptides to be quickly discovered and characterized from the cultures of three strains of Bifidobacterium, B. animalis, B. longum, and B. pseudolongum. Interestingly, four were uniquely secreted by B. animalis treated with gypenosides, and one showed ACE inhibitory effect with an IC₅₀ value of 193.22 μM. Consequently, this approach could serve as a powerful tool for quickly discovering bioactive peptides from the complex metabolites of probiotics, which contributes to the development of functional foods and nutraceuticals.

Keywords: Bifidobacterium; Bioactive peptides; Feature-based molecular networking; MCX coupled with LC–MS.

MeSH terms

Bifidobacterium / metabolism*
Chromatography, Liquid / methods*
Mass Spectrometry / methods*
Peptides / analysis*
Peptides / isolation & purification
Probiotics
Solid Phase Extraction / methods*

Substances

Peptides