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Over-expression of native Saccharomyces cerevisiae exocytic SNARE genes increased heterologous cellulase secretion

  • Applied genetics and molecular biotechnology
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Abstract

SNAREs (soluble NSF [N-ethylmaleimide-sensitive factor] attachment receptor proteins) are required at the majority of fusion events during intracellular membrane transport and play crucial roles in facilitating protein trafficking between the various membrane-enclosed organelles and the plasma membrane. We demonstrate increases in the secretion of the Talaromyces emersonii Cel7A (a cellobiohydrolase) and the Saccharomycopsis fibuligera Cel3A (a β-glucosidase), through the separate and simultaneous over-expression of different components of the exocytic SNARE complex in Saccharomyces cerevisiae. Over-expression of SNC1 yielded the biggest improvement in Te-Cel7A secretion (71 %), whilst SSO1 over-expression lead to the highest increases in Sf-Cel3A secretion (43.8 %). Simultaneous over-expression of differential combinations of these SNARE components yielded maximal increases of ~52 % and ~49 % for the secretion of Te-Cel7A and Sf-Cel3A, respectively. These increases generally did not cause deleterious growth effects, whilst differential improvement patterns were observed for the two reporter proteins (Sf-Cel3A and Te-Cel7A). Simultaneous over-expression of up to three of these components, in strains secreting the more efficiently expressed Sf-Cel3A, illustrated a slight decrease in osmotic tolerance at elevated NaCl concentrations, as well as a detectable decrease in ethanol tolerance at increased concentrations. This work illustrates the potential of engineering components of the anterograde secretory pathway, particularly its SNARE components, for the improvement of heterologous cellulase secretion.

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Acknowledgements

Funding for this project was provided by the National Research Foundation (South Africa).

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The authors declare that there are no conflicts of interest associated with the submission of this study.

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  1. Department of Microbiology, Stellenbosch University, Stellenbosch, 7602, South Africa

    John Henry D. Van Zyl, Riaan Den Haan & Willem H. Van Zyl

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  1. John Henry D. Van Zyl
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  2. Riaan Den Haan
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  3. Willem H. Van Zyl
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Correspondence to Willem H. Van Zyl.

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Van Zyl, J.H.D., Den Haan, R. & Van Zyl, W.H. Over-expression of native Saccharomyces cerevisiae exocytic SNARE genes increased heterologous cellulase secretion. Appl Microbiol Biotechnol 98, 5567–5578 (2014). https://doi.org/10.1007/s00253-014-5647-1

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