Open Access Highly Accessed Research article

Myosin-cross-reactive antigen (MCRA) protein from Bifidobacterium breve is a FAD-dependent fatty acid hydratase which has a function in stress protection

Eva Rosberg-Cody124, Alena Liavonchanka3, Cornelia Göbel3, R Paul Ross14, Orla O'Sullivan4, Gerald F Fitzgerald12, Ivo Feussner3 and Catherine Stanton14*

Author Affiliations

1 Alimentary Pharmabiotic Centre, Cork, Ireland

2 Microbiology Department, University College Cork, Cork, Ireland

3 Department Plant Biochemistry, Albrecht-von-Haller-Institute, Georg-August-University Göttingen, Göttingen, Germany

4 Teagasc Food Research Centre, Moorepark, Fermoy, Co. Cork, Ireland

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BMC Biochemistry 2011, 12:9  doi:10.1186/1471-2091-12-9

Published: 17 February 2011

Additional files

Additional file 1:

Map of putative isomerase gene. Map of putative isomerase gene and chromosome walking strategy (inverse PCR) used to obtain the full sequence.

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Additional file 2:

Mcra gene sequence data from Bifidobacterium strains and Propionibacterium freudenreichii shermanii. 1 kb PCR fragments following PCR of genomic DNA from a range of bifidobacterial strains and Propionibacterium freudenreichii shermanii 9093 using oligonucleotide primers EV1a forward and EV2a reverse. Lanes 1-12 in the following order; B. adolescentis; B. breve 2257; B. breve 702258; B. breve 8815; B. dentium 2243; B. infantis 2205; B. lactis Bb 12; B. longum 2259; B. longum BB 536; Bifidobacterium sp. 35612; Bifidobacterium sp. 35687; Propionibacterium freudenreichii shermanii 9093.

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Additional file 3:

Putative linoleic acid isomerase protein. Panel A. E. coli JM109(DE3) pOXO4 (lanes 1-4) and pEMYE3 (lanes 5- 8) induced with 0.5 mM IPTG. Samples taken after 1, 3, 6 and 12 h growth for protein extraction and SDS-PAGE. Panel B. C. glutamicum pCLIK (lane 1) and pMYC2 (lane 2) following 24 h growth prior to protein extraction. Panel C. L. lactis pNZ4848 (lanes 1-2) and pEMYL1 (lanes 3-4) following induction with nisin 0 and 10 ng/ml, respectively, and further growth for 12 h prior to protein extraction.

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Additional file 4:

CLA forming activity in bacterial strains expressing MCRA from B. breve. GC chromatogram of supernatant from B. breve NCIMB 702258 (positive control), L. lactis pEMYL1 (carrying laiA gene), and L. lactis pNZ8048 (vector control) following growth in linoleic acid (0.1 mg/ml) (L. lactis) and (0.5 mg/ml (B. breve) for 48-72 h. Similar results were obtained for recombinant C. glutamicum and E. coli.

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Additional file 5:

Hydratase activity in bacterial strains expressing MCRA from B. breve. Amounts of 10-HOA (black bars) and 10-HOE (gray bars) in supernatant from L. lactis pEMYL1 (carrying MCRA gene), and L. lactis pNZ8048 (VC, vector control) following growth in linoleic acid (0.1 mg/ml) (L. lactis) for 2-30 h. Data represent the mean values of 3 measurements. Similar results were obtained for recombinant C. glutamicum.

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