Feruloyl Esterase Protocols
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Introduction
These are methods to screen for and assay Ferulic-acid Esterase activity. While the screening method outlined here is specific for Lactobacillus 'species' it can be modified for other species by changing the media type to one appropriate for that species.
Plate Screen
Materials
- Ethyl ferulate stock solution (10%v/v in dimethylformamide)
- MRS agar
- If screening natural strains some find it helpfull to eliminate glucose from the media to drive FAE secretion
- Petri plates
Method
- Prepare 300ml of MRS media.
- Autoclave the media for 20min on liquid cycle.
- Let the autoclaved media cool in a 65°C water bath for 1 hour.
- Add 300μL ethyl ferulate to 2.7ml dimethylformamide.
Assay
Materials
Method
Notes
References
Donaghy, J., P. F. Kelly, et al. (1998). "Detection of ferulic acid esterase production by Bacillus spp. and lactobacilli." Applied Microbiology and Biotechnology 50(2): 257-260.
Mastihuba, V., L. Kremnicky, et al. (2002). "A spectrophotometric assay for feruloyl esterases." Analytical Biochemistry 309(1): 96-101.
Nsereko, V. L., B. K. Smiley, et al. (2008). "Influence of inoculating forage with lactic acid bacterial strains that produce ferulate esterase on ensilage and ruminal degradation of fiber." Animal Feed Science and Technology 145(1-4): 122-135.
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