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The Protein Journal

, Volume 34, Issue 2, pp 135–146 | Cite as

Identification of Proteins Possibly Involved in Glucosinolate Metabolism in L. agilis R16 and E. coli VL8

  • Vijitra Luang-In
  • Arjan Narbad
  • Fatma Cebeci
  • Mark Bennett
  • John T. Rossiter
Article

Abstract

This study was aimed to identify sinigrin-induced bacterial proteins potentially involved in the metabolism of glucosinolate in two glucosinolate-metabolising bacteria Lactobacillus agilis R16 and Escherichia coli VL8. Sinigrin (2 mM) was used to induce the proteins in both bacteria under anaerobic incubation for 8 h at 30 °C for L. agilis R16 and 37 °C for E. coli VL8 and the controls without sinigrin were performed. Allyl isothiocyanate and allyl nitrile as two degradation products of sinigrin were detected in sinigrin-induced cultures of L. agilis R16 (27 % total products) and E. coli VL8 (38 % total products) from a complete sinigrin degradation in 8 h for both bacteria. 2D gel electrophoresis was conducted to identify induced proteins with at least twofold increased abundance. Sinigrin-induced L. agilis R16 and the control produced 1561 and 1543 protein spots, respectively. For E. coli VL8, 1363 spots were detected in sinigrin-induced and 1354 spots in the control. A combination of distinct proteins and upregulated proteins of 32 and 35 spots in L. agilis R16 and E. coli VL8, respectively were detected upon sinigrin induction. Of these, 12 and 16 spots from each bacterium respectively were identified by LC–MS/MS. In both bacteria most of the identified proteins are involved in carbohydrate metabolism, oxidoreduction system and sugar transport while the minority belong to purine metabolism, hydrolysis, and proteolysis. This indicated that sinigrin induction led to the expressions of proteins with similar functions in both bacteria and these proteins may play a role in bacterial glucosinolate metabolism.

Keywords

Glucosinolate Gut bacteria Myrosinase Isothiocyanate Two-dimensional electrophoresis 

Abbreviations

GSL

Glucosinolate

ITC

Isothiocyanate

NIT

Nitriles

2D

Two dimension

LA

L. agilis R16

ECO

E. coli VL8

LC–MS/MS

Liquid chromatography mass spectrometry

Notes

Acknowledgments

The authors thank Llanos-Palop et al. (1995) for L. agilis R16 strain used in this work. The study was supported in part by a strategic programme grant to Institute of Food Research (Norwich) from the UK Biotechnology and Biological Sciences Research Council (BB/J004545/1).

Conflict of interest

The authors have declared no conflict of interest.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Vijitra Luang-In
    • 1
    • 4
  • Arjan Narbad
    • 2
  • Fatma Cebeci
    • 3
  • Mark Bennett
    • 4
  • John T. Rossiter
    • 4
  1. 1.Natural Antioxidant Research Unit, Department of Biotechnology, Faculty of TechnologyMahasarakham UniversityMahasarakhamThailand
  2. 2.Gut Health and Food Safety ProgrammeInstitute of Food ResearchNorwichUK
  3. 3.Food and Health ProgrammeInstitute of Food ResearchNorwichUK
  4. 4.Faculty of Life SciencesImperial College LondonLondonUK

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