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A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability

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Abstract

The aim of this work was to construct a novel food-grade industrial arming yeast displaying β-1,3-1,4-glucanase and to evaluate the thermal stability of the glucanase for practical application. For this purpose, a bi-directional vector containing galactokinase (GAL1) and phosphoglycerate kinase 1 (PGK1) promoters in different orientations was constructed. The β-1,3-1,4-glucanase gene from Bacillus subtilis was fused to α-agglutinin and expressed under the control of the GAL1 promoter. α-galactosidase induced by the constitutive PGK1 promoter was used as a food-grade selection marker. The feasibility of the α-galactosidase marker was confirmed by the growth of transformants harboring the constructed vector on a medium containing melibiose as a sole carbon source, and by the clear halo around the transformants in Congo-red plates owing to the expression of β-1,3-1,4-glucanase. The analysis of β-1,3-1,4-glucanase activity in cell pellets and in the supernatant of the recombinant yeast strain revealed that β-1,3-1,4-glucanase was successfully displayed on the cell surface of the yeast. The displayed β-1,3-1,4-glucanase activity in the recombinant yeast cells increased immediately after the addition of galactose and reached 45.1 U/ml after 32-h induction. The thermal stability of β-1,3-1,4-glucanase displayed in the recombinant yeast cells was enhanced compared with the free enzyme. These results suggest that the constructed food-grade yeast has the potential to improve the brewing properties of beer.

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Authors and Affiliations

  1. Department of Food Science and Nutrition, Zhejiang University, Hangzhou, 310029, China

    Qin Guo, Liu-Liu Ma, Qi-He Chen, Ji-Cheng Chen, Hong-Bo Zhang, Hui Ruan & Guo-Qing He

  2. Wenzhou Medical College, Wenzhou, 325035, China

    Wei Zhang

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  1. Qin Guo
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  2. Wei Zhang
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  3. Liu-Liu Ma
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  4. Qi-He Chen
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  5. Ji-Cheng Chen
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  6. Hong-Bo Zhang
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  7. Hui Ruan
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  8. Guo-Qing He
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Correspondence to Hui Ruan or Guo-Qing He.

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Project (No. 2006AA10Z316) supported by the Hi-Tech Research and Development Program (863) of China

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Guo, Q., Zhang, W., Ma, LL. et al. A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability. J. Zhejiang Univ. Sci. B 11, 41–51 (2010). https://doi.org/10.1631/jzus.B0900185

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