scholarly journals Expression cassette and plasmid construction for Yeast Surface Display in Saccharomyces cerevisiae

2021 ◽  
Author(s):  
Renan Eugênio Araujo Piraine ◽  
Vitória Sequeira Gonçalves ◽  
Alceu Gonçalves dos Santos Junior ◽  
Rodrigo Casquero Cunha ◽  
Pedro Machado Medeiros de Albuquerque ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Surface Display ◽  
Yeast Surface Display ◽  
Expression Cassette ◽  
Plasmid Construction ◽  
Yeast Surface

scholarly journals Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis

Catalysts ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 757
Author(s):  
Huiyi Shang ◽  
Danni Yang ◽  
Dairong Qiao ◽  
Hui Xu ◽  
Yi Cao
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Weight ◽  
Large Scale ◽  
Surface Display ◽  
Yeast Surface Display ◽  
Fusion Partner ◽  
Whole Cell ◽  
Food Industries ◽  
Yeast Surface ◽  
The Stability

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.

scholarly journals Directed Evolution of Brain-Derived Neurotrophic Factor for Improved Folding and Expression in Saccharomyces cerevisiae

2014 ◽  
Vol 80 (18) ◽  
pp. 5732-5742 ◽  
Author(s):  
Michael L. Burns ◽  
Thomas M. Malott ◽  
Kevin J. Metcalf ◽  
Benjamin J. Hackel ◽  
Jonah R. Chan ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Directed Evolution ◽  
Neurotrophic Factor ◽  
Surface Display ◽  
Brain Derived Neurotrophic Factor ◽  
Binding Activity ◽  
Yeast Surface Display ◽  
Site Directed Mutagenesis ◽  
Secretion Systems ◽  
Yeast Surface

ABSTRACTBrain-derived neurotrophic factor (BDNF) plays an important role in nervous system function and has therapeutic potential. Microbial production of BDNF has resulted in a low-fidelity protein product, often in the form of large, insoluble aggregates incapable of binding to cognate TrkB or p75 receptors. In this study, employingSaccharomyces cerevisiaedisplay and secretion systems, it was found that BDNF was poorly expressed and partially inactive on the yeast surface and that BDNF was secreted at low levels in the form of disulfide-bonded aggregates. Thus, for the purpose of increasing the compatibility of yeast as an expression host for BDNF, directed-evolution approaches were employed to improve BDNF folding and expression levels. Yeast surface display was combined with two rounds of directed evolution employing random mutagenesis and shuffling to identify BDNF mutants that had 5-fold improvements in expression, 4-fold increases in specific TrkB binding activity, and restored p75 binding activity, both as displayed proteins and as secreted proteins. Secreted BDNF mutants were found largely in the form of soluble homodimers that could stimulate TrkB phosphorylation in transfected PC12 cells. Site-directed mutagenesis studies indicated that a particularly important mutational class involved the introduction of cysteines proximal to the native cysteines that participate in the BDNF cysteine knot architecture. Taken together, these findings show that yeast is now a viable alternative for both the production and the engineering of BDNF.

Application of yeast surface display system in expression of recombinant pediocin PA-1 in Saccharomyces cerevisiae

2020 ◽  
Vol 65 (6) ◽  
pp. 955-961
Author(s):  
Thu Pham Anh Nguyen ◽  
Thu Thi Minh Nguyen ◽  
Nghia Hieu Nguyen ◽  
Tri Nhan Nguyen ◽  
Thao Thi Phuong Dang
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Surface Display ◽  
Yeast Surface Display ◽  
Display System ◽  
Yeast Surface

Isomaltulose production via yeast surface display of sucrose isomerase from Enterobacter sp. FMB-1 on Saccharomyces cerevisiae

2011 ◽  
Vol 102 (19) ◽  
pp. 9179-9184 ◽  
Author(s):  
Gil-Yong Lee ◽  
Jong-Hyun Jung ◽  
Dong-Ho Seo ◽  
Jantra Hansin ◽  
Suk-Jin Ha ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Surface Display ◽  
Yeast Surface Display ◽  
Yeast Surface ◽  
Sucrose Isomerase

scholarly journals The Role of Yeast-Surface-Display Techniques in Creating Biocatalysts for Consolidated BioProcessing

Catalysts ◽  
2018 ◽  
Vol 8 (3) ◽  
pp. 94 ◽  
Author(s):  
Ian Dominic Flormata Tabañag ◽  
I-Ming Chu ◽  
Yu-Hong Wei ◽  
Shen-Long Tsai
Keyword(s):  
Climate Change ◽  
Lignocellulosic Biomass ◽  
Surface Engineering ◽  
Consolidated Bioprocessing ◽  
Surface Display ◽  
Yeast Surface Display ◽  
Qualitative Assessment ◽  
The Status ◽  
Yeast Surface

Climate change is directly linked to the rapid depletion of our non-renewable fossil resources and has posed concerns on sustainability. Thus, imploring the need for us to shift from our fossil based economy to a sustainable bioeconomy centered on biomass utilization. The efficient bioconversion of lignocellulosic biomass (an ideal feedstock) to a platform chemical, such as bioethanol, can be achieved via the consolidated bioprocessing technology, termed yeast surface engineering, to produce yeasts that are capable of this feat. This approach has various strategies that involve the display of enzymes on the surface of yeast to degrade the lignocellulosic biomass, then metabolically convert the degraded sugars directly into ethanol, thus elevating the status of yeast from an immobilization material to a whole-cell biocatalyst. The performance of the engineered strains developed from these strategies are presented, visualized, and compared in this article to highlight the role of this technology in moving forward to our quest against climate change. Furthermore, the qualitative assessment synthesized in this work can serve as a reference material on addressing the areas of improvement of the field and on assessing the capability and potential of the different yeast surface display strategies on the efficient degradation, utilization, and ethanol production from lignocellulosic biomass.

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