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Journal of Zhejiang University SCIENCE B 2010 Vol.11 No.1 P.41-51

10.1631/jzus.B0900185


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


Author(s):  Qin GUO, Wei ZHANG, Liu-liu MA, Qi-he CHEN, Ji-cheng CHEN, Hong-bo ZHANG, Hui RUAN, Guo-qing HE

Affiliation(s):  Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China; more

Corresponding email(s):   ruanhui98@yahoo.com.cn, gqhe@zju.edu.cn

Key Words:  &alpha, -agglutinin, Food-grade selection marker, &beta, -1, 3-1, 4-glucanase, &alpha, -galactosidase, Thermostability


Qin GUO, Wei ZHANG, Liu-liu MA, Qi-he CHEN, Ji-cheng CHEN, Hong-bo ZHANG, Hui RUAN, Guo-qing HE. A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability[J]. Journal of Zhejiang University Science B, 2010, 11(1): 41-51.

@article{title="A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability",
author="Qin GUO, Wei ZHANG, Liu-liu MA, Qi-he CHEN, Ji-cheng CHEN, Hong-bo ZHANG, Hui RUAN, Guo-qing HE",
journal="Journal of Zhejiang University Science B",
volume="11",
number="1",
pages="41-51",
year="2010",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B0900185"
}

%0 Journal Article
%T A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability
%A Qin GUO
%A Wei ZHANG
%A Liu-liu MA
%A Qi-he CHEN
%A Ji-cheng CHEN
%A Hong-bo ZHANG
%A Hui RUAN
%A Guo-qing HE
%J Journal of Zhejiang University SCIENCE B
%V 11
%N 1
%P 41-51
%@ 1673-1581
%D 2010
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0900185

TY - JOUR
T1 - A food-grade industrial arming yeast expressing β-1,3-1,4-glucanase with enhanced thermal stability
A1 - Qin GUO
A1 - Wei ZHANG
A1 - Liu-liu MA
A1 - Qi-he CHEN
A1 - Ji-cheng CHEN
A1 - Hong-bo ZHANG
A1 - Hui RUAN
A1 - Guo-qing HE
J0 - Journal of Zhejiang University Science B
VL - 11
IS - 1
SP - 41
EP - 51
%@ 1673-1581
Y1 - 2010
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0900185


Abstract: 
The aim of this work was to construct a novel food-grade industrial arming yeast displaying &beta;-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 &beta;-1,3-1,4-glucanase gene from Bacillus subtilis was fused to &alpha%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>&alpha;-agglutinin and expressed under the control of the GAL1 promoter. &alpha%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>&alpha;-galactosidase induced by the constitutive PGK1 promoter was used as a food-grade selection marker. The feasibility of the &alpha%29&ck%5B%5D=abstract&ck%5B%5D=keyword'>&alpha;-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 &beta;-1,3-1,4-glucanase. The analysis of &beta;-1,3-1,4-glucanase activity in cell pellets and in the supernatant of the recombinant yeast strain revealed that &beta;-1,3-1,4-glucanase was successfully displayed on the cell surface of the yeast. The displayed &beta;-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 &beta;-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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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