scholarly journals Display of Microbial Glucose Dehydrogenase and Cholesterol Oxidase on the Yeast Cell Surface for the Detection of Blood Biochemical Parameters

Biosensors ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 13
Author(s):  
Shiyao Zhao ◽  
Dong Guo ◽  
Quanchao Zhu ◽  
Weiwang Dou ◽  
Wenjun Guan
Keyword(s):  
Blood Glucose ◽  
Cholesterol Oxidase ◽  
Glucose Dehydrogenase ◽  
Surface Display ◽  
Storage Period ◽  
Glucose Biosensor ◽  
Yeast Surface Display ◽  
Yeast Cells ◽  
Yeast Cell Surface ◽  
Blood Biochemical

High levels of blood glucose are always associated with numerous complications including cholesterol abnormalities. Therefore, it is important to simultaneously monitor blood glucose and cholesterol levels in patients with diabetes during the management of chronic diseases. In this study, a glucose dehydrogenase from Aspergillus oryzae TI and a cholesterol oxidase from Chromobacterium sp. DS-1 were displayed on the surface of Saccharomyces cerevisiae, respectively, using the yeast surface display system at a high copy number. In addition, two whole-cell biosensors were constructed through the immobilization of the above yeast cells on electrodes, for electrochemical detection of glucose and cholesterol. The assay time was 8.5 s for the glucose biosensors and 30 s for the cholesterol biosensors. Under optimal conditions, the cholesterol biosensor exhibited a linear range from 2 to 6 mmol·L−1. The glucose biosensor responded efficiently to the presence of glucose at a concentration range of 20–600 mg·dL−1 (1.4–33.3 mmol·L−1) and showed excellent anti-xylose interference properties. Both biosensors exhibited good performance at room temperature and remained stable over a three-week storage period.

Microbial surface display of glucose dehydrogenase for amperometric glucose biosensor

2013 ◽  
Vol 45 ◽  
pp. 19-24 ◽  
Author(s):  
Bo Liang ◽  
Liang Li ◽  
XiangJiang Tang ◽  
Qiaolin Lang ◽  
Hongwei Wang ◽  
...  
Keyword(s):  
Glucose Dehydrogenase ◽  
Surface Display ◽  
Glucose Biosensor

scholarly journals Yeast Surface Display of Trifunctional Minicellulosomes for Simultaneous Saccharification and Fermentation of Cellulose to Ethanol

2009 ◽  
Vol 76 (4) ◽  
pp. 1251-1260 ◽  
Author(s):  
Fei Wen ◽  
Jie Sun ◽  
Huimin Zhao
Keyword(s):  
Cell Surface ◽  
Consolidated Bioprocessing ◽  
Surface Display ◽  
Cellulose Hydrolysis ◽  
Cellulase Production ◽  
Single Step ◽  
Yeast Surface Display ◽  
Biofuel Production ◽  
Yeast Cells ◽  
Recombinant Yeast Strain

ABSTRACT By combining cellulase production, cellulose hydrolysis, and sugar fermentation into a single step, consolidated bioprocessing (CBP) represents a promising technology for biofuel production. Here we report engineering of Saccharomyces cerevisiae strains displaying a series of uni-, bi-, and trifunctional minicellulosomes. These minicellulosomes consist of (i) a miniscaffoldin containing a cellulose-binding domain and three cohesin modules, which was tethered to the cell surface through the yeast a-agglutinin adhesion receptor, and (ii) up to three types of cellulases, an endoglucanase, a cellobiohydrolase, and a β-glucosidase, each bearing a C-terminal dockerin. Cell surface assembly of the minicellulosomes was dependent on expression of the miniscaffoldin, indicating that formation of the complex was dictated by the high-affinity interactions between cohesins and dockerins. Compared to the unifunctional and bifunctional minicellulosomes, the quaternary trifunctional complexes showed enhanced enzyme-enzyme synergy and enzyme proximity synergy. More importantly, surface display of the trifunctional minicellulosomes gave yeast cells the ability to simultaneously break down and ferment phosphoric acid-swollen cellulose to ethanol with a titer of ∼1.8 g/liter. To our knowledge, this is the first report of a recombinant yeast strain capable of producing cell-associated trifunctional minicellulosomes. The strain reported here represents a useful engineering platform for developing CBP-enabling microorganisms and elucidating principles of cellulosome construction and mode of action.

scholarly journals Yeast Surface Display System: Strategies for Improvement and Biotechnological Applications

2022 ◽  
Vol 9 ◽  
Author(s):  
Karla V. Teymennet-Ramírez ◽  
Fernando Martínez-Morales ◽  
María R. Trejo-Hernández
Keyword(s):  
Protein Interactions ◽  
Genetic Manipulation ◽  
Surface Display ◽  
Yeast Surface Display ◽  
Yeast Cells ◽  
Biotechnological Applications ◽  
Protein Protein Interactions ◽  
Post Translational Modifications ◽  
Yeast Surface ◽  
Antibody Design

Yeast surface display (YSD) is a “whole-cell” platform used for the heterologous expression of proteins immobilized on the yeast’s cell surface. YSD combines the advantages eukaryotic systems offer such as post-translational modifications, correct folding and glycosylation of proteins, with ease of cell culturing and genetic manipulation, and allows of protein immobilization and recovery. Additionally, proteins displayed on the surface of yeast cells may show enhanced stability against changes in temperature, pH, organic solvents, and proteases. This platform has been used to study protein-protein interactions, antibody design and protein engineering. Other applications for YSD include library screening, whole-proteome studies, bioremediation, vaccine and antibiotics development, production of biosensors, ethanol production and biocatalysis. YSD is a promising technology that is not yet optimized for biotechnological applications. This mini review is focused on recent strategies to improve the efficiency and selection of displayed proteins. YSD is presented as a cutting-edge technology for the vectorial expression of proteins and peptides. Finally, recent biotechnological applications are summarized. The different approaches described herein could allow for a better strategy cascade for increasing protein/peptide interaction and production.

Expression Cassette and Plasmid Construction for Yeast Surface Display in Saccharomyces Cerevisiae

2021 ◽  
Author(s):  
Renan E A Piraine ◽  
Vitória S Gonçalves ◽  
Alceu GS dos Santos Junior ◽  
Rodrigo C Cunha ◽  
Pedro MM Albuquerque ◽  
...  
Keyword(s):  
Cell Surface ◽  
Recombinant Protein Expression ◽  
Polyclonal Antibodies ◽  
Surface Display ◽  
Cell Surface Display ◽  
Yeast Surface Display ◽  
Expression Cassette ◽  
Dot Blot ◽  
Herpesvirus Type ◽  
Yeast Cell Surface

Abstract Objectives. Develop a Cell Surface Display system in S. cerevisiae, based on the construction of an expression cassette for pYES2 plasmid. Results. The construction of an expression cassette containing the α-factor signal peptide and the C-terminal portion of the α-agglutinin protein was made and its sequence inserted into a plasmid named pYES2/gDαAgglutinin, allowing cell surface display of bovine herpesvirus type 5 (BoHV-5) glycoprotein D (gD) on S. cerevisiae BY4741 strain. Recombinant protein expression was confirmed by dot blot, and indirect immunofluorescence using monoclonal anti-histidine antibodies and polyclonal antibodies from mice experimentally vaccinated with a recombinant gD. Conclusions. These results demonstrate that the approach and plasmid used represent not only an effective system for immobilizing proteins on the yeast cell surface, as well as a platform for immunobiologicals development.

Detection of Glucose in Human Serum Based on Silicon Dot Probe

2020 ◽  
Vol 16 (6) ◽  
pp. 744-752
Author(s):  
Kuan Luo ◽  
Xinyu Jiang
Keyword(s):  
Quantum Dots ◽  
Blood Glucose ◽  
Human Serum ◽  
Organic Dyes ◽  
Limit Of Detection ◽  
Fasting Blood Glucose ◽  
Glucose Biosensor ◽  
Metal Nanoclusters ◽  
Glucose Levels ◽  
Highly Sensitive

Background: Diabetes Mellitus (DM) is a major public metabolic disease that influences 366 million people in the world in 2011, and this number is predicted to rise to 552 million in 2030. DM is clinically diagnosed by a fasting blood glucose that is equal or greater than 7 mM. Therefore, the development of effective glucose biosensor has attracted extensive attention worldwide. Fluorescence- based strategies have sparked tremendous interest due to their rapid response, facile operation, and excellent sensitivity. Many fluorescent compounds have been employed for precise analysis of glucose, including quantum dots, noble metal nanoclusters, up-converting nanoparticles, organic dyes, and composite fluorescent microspheres. Silicon dot as promising quantum dots materials have received extensive attention, owing to their distinct advantages such as biocompatibility, low toxicity and high photostability. Methods: MnO2 nanosheets on the Si nanoparticles (NPs) surface serve as a quencher. Si NPs fluorescence can make a recovery by the addition of H2O2, which can reduce MnO2 to Mn2+, and the glucose can thus be monitored based on the enzymatic conversion of glucose by glucose oxidase to generate H2O2. Therefore, the glucose concentration can be derived by recording the fluorescence recovery spectra of the Si NPs. Results: This probe enabled selective detection of glucose with a linear range of 1-100 μg/mL and a limit of detection of 0.98 μg/mL. Compared with the commercial glucometer, this method showed favorable results and convincing reliability. Conclusion: We have developed a novel method based on MnO2 -nanosheet-modified Si NPs for rapid monitoring of blood glucose levels. By combining the highly sensitive H2O2/MnO2 reaction with the excellent photostability of Si NPs, a highly sensitive, selective, and cost-efficient sensing approach for glucose detection has been designed and applied to monitor glucose levels in human serum with satisfactory results.

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.

Effect of Estrus Induction on Blood Biochemical and Mineral Profiles and Fertility Rate in Retained Fetal Membranes Affected and Normally Calved Riverine Buffaloes

2021 ◽  
Author(s):  
M. Selvaraju ◽  
K. Ganesh
Keyword(s):  
Blood Glucose ◽  
Total Protein ◽  
Fertility Rate ◽  
Fetal Membranes ◽  
Blood Biochemical ◽  
Group I ◽  
Mineral Mixture ◽  
Calcium Phosphorus ◽  
Calcium And Phosphorus ◽  
Phosphorus Levels

Background: Calving to conception interval is abnormally extended by the occurrence of retained fetal membranes (RFM) by altering the blood biochemical and mineral milieu in cows. Hence this experiment was conducted in RFM affected and normally calved (NC) buffaloes by inducing estrus with CIDR plus PGF2α protocol to study the blood biochemical and mineral profiles and to correlate them with fertility rate. Methods: Buffaloes (n=64) at 45-60 days postpartum including 32 treated for RFM and 32 NC from field were equally divided into groups I and II and groups III and IV, respectively. Buffaloes of groups I and III were initially dewormed and administered 35-50 g mineral mixture daily orally for 15 days in the concentrate feed. Then, buffaloes of all the groups were treated with CIDR plus PGF2α. After CIDR removal, all the buffaloes were artificially inseminated twice at 48 and 72 hrs. Blood was collected during different stages of treatment from all the buffaloes to assess the blood biochemical and mineral status. The animals returned to estrus following FTAI were again inseminated during subsequent estrus. Pregnancy diagnosis was done at 60 days post-AI and conception rates for induced estrus and overall of two cycles were calculated.Result: In all the groups, blood glucose, total protein, triglycerides, cholesterol and phosphorus levels increased from the time of selection to 10 days post-AI. There was an altered calcium phosphorus ratio in RFM affected buffaloes (1:1) at the time of selection. CIDR plus PGF2α protocol influenced the blood biochemical constituents and brought the calcium and phosphorus ratio as 2:1 and improved the fertility in riverine buffaloes. The overall conception rate was 62.50, 37.50, 87.50 and 75.00 per cent, in group I, II, III and IV buffaloes, respectively.

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