Protein engineering of invertase for enhancing yeast dough fermentation under high-sucrose conditions
Journal article, 2023

During yeast dough fermentation, such as the high-sucrose bread-making process, the yeast cells are subjected to considerable osmotic stress, resulting in poor outcomes. Invertase is important for catalyzing the irreversible hydrolysis of sucrose to free glucose and fructose, and decreasing the catalytic activity of the invertase may reduce the glucose osmotic stress on the yeast. In this study, we performed structural design and site-directed mutagenesis (SDM) on the Saccharomyces cerevisiae invertase (ScInV) in an Escherichia coli expression system to study the catalytic activity of ScInV mutants in vitro. In addition, we generated the same mutation sites in the yeast endogenous genome and tested their invertase activity in yeast and dough fermentation ability. Our results indicated that appropriately reduced invertase activity of yeast ScInV can enhance dough fermentation activity under high-sucrose conditions by 52%. Our systems have greatly accelerated the engineering of yeast endogenous enzymes both in vitro and in yeast, and shed light on future metabolic engineering of yeast.

Site-directed mutagenesis

Invertase activity

Fermentation ability

Protein engineering


Yijin Zhao

Beijing University of Chemical Technology

China Agricultural University

Kaiwen Meng

China Agricultural University

Jinyu Fu

Beijing University of Chemical Technology

Shijie Xu

Beijing University of Chemical Technology

Guang Cai

Beijing University of Chemical Technology

Geng Meng

China Agricultural University

Jens B Nielsen

Beijing University of Chemical Technology

Chalmers, Life Sciences, Systems and Synthetic Biology

Zihe Liu

Beijing University of Chemical Technology

Yueping Zhang

China Agricultural University

Folia Microbiologica

0015-5632 (ISSN) 18749356 (eISSN)

Vol. 68 2 207-217

Subject Categories

Biochemistry and Molecular Biology

Biocatalysis and Enzyme Technology

Other Industrial Biotechnology



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Latest update

7/5/2023 9