Surface Display of Malolactic Enzyme from Oenococcus oeni on Saccharomyces cerevisiae

2013 ◽  
Vol 169 (8) ◽  
pp. 2350-2361 ◽  
Author(s):  
Xiuyan Zhang ◽  
Xiaoyan Hou ◽  
Fang Liang ◽  
Fusheng Chen ◽  
Xiaohong Wang
Catalysts ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 757
Author(s):  
Huiyi Shang ◽  
Danni Yang ◽  
Dairong Qiao ◽  
Hui Xu ◽  
Yi Cao

Levan has wide applications in chemical, cosmetic, pharmaceutical and food industries. The free levansucrase is usually used in the biosynthesis of levan, but the poor reusability and low stability of free levansucrase have limited its large-scale use. To address this problem, the surface-displayed levansucrase in Saccharomyces cerevisiae were generated and evaluated in this study. The levansucrase from Zymomonas mobilis was displayed on the cell surface of Saccharomyces cerevisiae EBY100 using a various yeast surface display platform. The N-terminal fusion partner is based on a-agglutinin, and the C-terminal one is Flo1p. The yield of levan produced by these two whole-cell biocatalysts reaches 26 g/L and 34 g/L in 24 h, respectively. Meanwhile, the stability of the surface-displayed levansucrases is significantly enhanced. After six reuses, these two biocatalysts retained over 50% and 60% of their initial activities, respectively. Furthermore, the molecular weight and polydispersity test of the products suggested that the whole-cell biocatalyst of levansucrase displayed by Flo1p has more potentials in the production of levan with low molecular weight which is critical in certain applications. In conclusion, our method not only enable the possibility to reuse the enzyme, but also improves the stability of the enzyme.


2019 ◽  
Vol 21 (7) ◽  
pp. 1795-1808 ◽  
Author(s):  
Gregory Guirimand ◽  
Kentaro Inokuma ◽  
Takahiro Bamba ◽  
Mami Matsuda ◽  
Kenta Morita ◽  
...  

Xylitol is a major commodity chemical widely used in both the food and pharmaceutical industries.


2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Xiaoyu Yang ◽  
Hongting Tang ◽  
Meihui Song ◽  
Yu Shen ◽  
Jin Hou ◽  
...  

Bioengineered ◽  
2013 ◽  
Vol 4 (3) ◽  
pp. 147-152 ◽  
Author(s):  
Christina Schümann ◽  
Herbert Michlmayr ◽  
Andrés M. del Hierro ◽  
Klaus D. Kulbe ◽  
Vladimir Jiranek ◽  
...  

2020 ◽  
Vol 8 (5) ◽  
pp. 655 ◽  
Author(s):  
Santiago Benito

The combined use of Lachancea thermotolerans and Schizosaccharomyces pombe is a new winemaking biotechnology that aims to solve some modern industrial oenology problems related to warm viticulture regions. These areas are characterized for producing musts with high levels of sugar that can potentially be converted into wines with elevated ethanol contents, which are usually associated with high pH levels. This biotechnology was reported for the first time in 2015, and since then, several scientific articles have been published regarding this topic. These reported scientific studies follow an evolution similar to that performed in the past for Saccharomyces cerevisiae and Oenococcus oeni; they start by reporting results for basic winemaking parameters at the beginning, later continuing with more advanced parameters. This review compares the results of different researchers that have applied this new biotechnology and have studied wine quality parameters such as ethanol, glycerol, malic acid, lactic acid, amino acids, aroma compounds, or anthocyanins. It is shown that the new biotechnology is repeatedly reported to solve specific winemaking problems such as the lack of acidity, biogenic amines, ethyl carbamate, or undesirable color losses. Such results highlight this biotechnology as a promising option for warm viticulture areas.


2006 ◽  
Vol 72 (11) ◽  
pp. 7140-7147 ◽  
Author(s):  
Frank Breinig ◽  
Björn Diehl ◽  
Sabrina Rau ◽  
Christian Zimmer ◽  
Helmut Schwab ◽  
...  

ABSTRACT Yeast cell surface display is a powerful tool for expression and immobilization of biocatalytically active proteins on a unicellular eukaryote. Here bacterial carboxylesterase EstA from Burkholderia gladioli was covalently anchored into the cell wall of Saccharomyces cerevisiae by in-frame fusion to the endogenous yeast proteins Kre1p, Cwp2p, and Flo1p. When p-nitrophenyl acetate was used as a substrate, the esterase specific activities of yeast expressing the protein fusions were 103 mU mg−1 protein for Kre1/EstA/Cwp2p and 72 mU mg−1 protein for Kre1/EstA/Flo1p. In vivo cell wall targeting was confirmed by esterase solubilization after laminarinase treatment and immunofluorescence microscopy. EstA expression resulted in cell wall-associated esterase activities of 2.72 U mg−1 protein for Kre1/EstA/Cwp2p and 1.27 U mg−1 protein for Kre1/EstA/Flo1p. Furthermore, esterase display on the yeast cell surface enabled the cells to effectively grow on the esterase-dependent carbon source glycerol triacetate (Triacetin). In the case of Kre1/EstA/Flo1p, in vivo maturation within the yeast secretory pathway and final incorporation into the wall were further enhanced when there was constitutive activation of the unfolded protein response pathway. Our results demonstrate that esterase cell surface display in yeast, which, as shown here, is remarkably more effective than EstA surface display in Escherichia coli, can be further optimized by activating the protein folding machinery in the eukaryotic secretion pathway.


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