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Metagenomic mining of feruloyl esterases from termite enteric flora

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Abstract

A metagenome expression library was created from Trinervitermes trinervoides termite hindgut symbionts and subsequently screened for feruloyl esterase (FAE) activities, resulting in seven recombinant fosmids conferring feruloyl esterase phenotypes. The amino acid sequence lengths of the seven FAE encoding open reading frames (ORFs) ranged from 260 to 274 aa and encoded polypeptides of between 28.9 and 31.4 kDa. The highest sequence identity scores for the seven ORFs against the GenBank database were between 45 and 59 % to a number of carboxyl ester hydrolyses. The seven FAE primary structures contained sequence motifs that correspond well with a classical pentapeptide (G-x-S-x-G) serine hydrolyse signature motif which harbours the catalytic serine residue in other FAE families. Six of the seven fae genes were successfully expressed heterologously in Escherichia coli, and the purified enzymes exhibited temperature optima range of 40–70 °C and the pH optima of between 6.5 and 8.0. The k cat/K M ratios for the six characterised FAEs showed the following order of substrate preference: methyl sinapate > methyl ferulate > ethyl ferulate. All six FAEs showed poor conversion rates against methyl p-coumarate and methyl caffeate, both of which lacked the methoxy (O–CH3) group substituent on the aromatic ring of the ester substrates, emphasising the requirement for at least one methoxy group on the aromatic ring of the hydroxycinnamic acid ester substrate for optimal FAE activity.

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Acknowledgments

The work was supported by the Technology Innovation Agency of South Africa and the CSIR South Africa. The authors would also like to thank Mr Harris Tshwane Manchidi, Johan Kemp and Dr Fritha Hennessey for the help with sample collection. Special thanks to Cornel Du Toit, UP Department of Entomology, University of Pretoria, for the help with termite collection and dissection and Dr Daniel Visser for his input during the screening programme.

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  1. CSIR Biosciences, Building 18 Meiring Naudé Road, Brummeria, PO Box 395, Pretoria, 0001, South Africa

    Konanani Rashamuse, Tina Ronneburg, Walter Sanyika, Kgama Mathiba, Edwin Mmutlane & Dean Brady

  2. Departments of Biotechnology and Food Technology, Tshwane University of Technology, Pretoria, 0001, South Africa

    Walter Sanyika & Dean Brady

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  1. Konanani Rashamuse
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  2. Tina Ronneburg
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Correspondence to Konanani Rashamuse.

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Konanani Rashamuse and Tina Ronneburg contributed equally to this work.

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Rashamuse, K., Ronneburg, T., Sanyika, W. et al. Metagenomic mining of feruloyl esterases from termite enteric flora. Appl Microbiol Biotechnol 98, 727–737 (2014). https://doi.org/10.1007/s00253-013-4909-7

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  • DOI: https://doi.org/10.1007/s00253-013-4909-7

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