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A multifunctional hybrid glycosyl hydrolase discovered in an uncultured microbial consortium from ruminant gut

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Abstract

A unique multifunctional glycosyl hydrolase was discovered by screening an environmental DNA library prepared from a microbial consortium collected from cow rumen. The protein consists of two adjacent catalytic domains. Sequence analysis predicted that one domain conforms to glycosyl hydrolase family 5 and the other to family 26. The enzyme is active on several different β-linked substrates and possesses mannanase, xylanase, and glucanase activities. Site-directed mutagenesis studies on the catalytic residues confirmed the presence of two functionally independent catalytic domains. Using site-specific mutations, it was shown that one catalytic site hydrolyzes β-1,4-linked mannan substrates, while the second catalytic site hydrolyzes β-1,4-linked xylan and β-1,4-linked glucan substrates. Polysaccharide Analysis using Carbohydrate gel Electrophoresis (PACE) also confirmed that the enzyme has discrete domains for binding and hydrolysis of glucan- and mannan-linked polysaccharides. Such multifunctional enzymes have many potential industrial applications in plant processing, including biomass saccharification, animal feed nutritional enhancement, textile, and pulp and paper processing.

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Acknowledgments

We gratefully acknowledge the contributions of Diversa colleagues Eric Mathur, Martin Keller, Xuqiu Tan, Mark Burk, Shaun Healey, Walt Callen, Lisa Bibbs, and the members of the Sequencing Group, Cathy Chang, and the members of the Library Construction Group.

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Authors and Affiliations

  1. Diversa Corporation, 4955 Directors Place, San Diego, CA, 92121, USA

    Nisha Palackal, Christopher S. Lyon, Seema Zaidi, Peter Luginbühl, John L. Macomber, Jay M. Short, Geoffrey P. Hazlewood, Dan E. Robertson & Brian A. Steer

  2. Department of Biochemistry, University of Cambridge, Cambridge, CB2 1QW, UK

    Paul Dupree & Florence Goubet

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  1. Nisha Palackal
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  2. Christopher S. Lyon
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  3. Seema Zaidi
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  4. Peter Luginbühl
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  5. Paul Dupree
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  6. Florence Goubet
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  7. John L. Macomber
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  8. Jay M. Short
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  9. Geoffrey P. Hazlewood
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  10. Dan E. Robertson
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  11. Brian A. Steer
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Correspondence to Brian A. Steer.

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Palackal, N., Lyon, C.S., Zaidi, S. et al. A multifunctional hybrid glycosyl hydrolase discovered in an uncultured microbial consortium from ruminant gut. Appl Microbiol Biotechnol 74, 113–124 (2007). https://doi.org/10.1007/s00253-006-0645-6

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  • DOI: https://doi.org/10.1007/s00253-006-0645-6

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