Transient Secretory Enzyme Expression in Leaf Protoplasts to Characterize SNARE Functional Classes in Conventional and Unconventional Secretion

2017 ◽  
pp. 209-221
Author(s):  
Di Sansebastiano Gian Pietro ◽  
Barozzi Fabrizio
Keyword(s):  
Enzyme Expression ◽  
Unconventional Secretion ◽  
Functional Classes ◽  
Secretory Enzyme ◽  
Leaf Protoplasts

Vascular Angiotensin-Converting Enzyme Expression Regulates Local Angiotensin II

Hypertension ◽  
1997 ◽  
Vol 29 (1) ◽  
pp. 98-104 ◽  
Author(s):  
Dominik N. Müller ◽  
Jürgen Bohlender ◽  
Karl F. Hilgers ◽  
Duska Dragun ◽  
Olivier Costerousse ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Angiotensin Converting Enzyme ◽  
Converting Enzyme ◽  
Enzyme Expression

scholarly journals Characterization and optimization of extracellular enzymes production by Aspergillus niger strains isolated from date by-products

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Reda Bellaouchi ◽  
Houssam Abouloifa ◽  
Yahya Rokni ◽  
Amina Hasnaoui ◽  
Nabil Ghabbour ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Culture Medium ◽  
Extracellular Enzymes ◽  
Enzyme Expression ◽  
Specific Enzyme ◽  
Enzymatic Production ◽  
High Production Rate ◽  
High Production ◽  
Incubation Temperatures ◽  
By Products

Abstract Background This work aims to study the optimal conditions of the fermentation culture medium used for the production of extracellular enzymes (amylase, cellulase, lipase, and protease) from previously isolated Aspergillus niger strains in date by-products. Results The five most powerful isolates selected based on the zone of degradation formed on Petri plates by the substrate were subjected to the quantitative evaluation of their enzymatic production. All five strains showed almost similar API-ZYM profiles, with minor variations observed at the level of some specific enzyme expression. The production of cellulase and amylase was depending on pH and incubation temperatures. ASP2 strain demonstrated the high production rate of amylase (at pH 5 and 30 °C) and cellulase (at pH 6 and 30 °C) for 96 h of incubation. Conclusion The A. niger showed the ability to produce several extracellular enzymes and can be used in the valorization of different agroindustrial residues.

Differential effects of inhibitors of PTZ‐induced kindling on glutamate transporters and enzyme expression

2021 ◽  
Author(s):  
Numan Taspinar ◽  
Ahmet Hacimuftuoglu ◽  
Selcuk Butuner ◽  
Basak Togar ◽  
Gokhan Arslan ◽  
...  
Keyword(s):  
Glutamate Transporters ◽  
Enzyme Expression ◽  
Differential Effects

scholarly journals Controlling Unconventional Secretion for Production of Heterologous Proteins in Ustilago maydis through Transcriptional Regulation and Chemical Inhibition of the Kinase Don3

2021 ◽  
Vol 7 (3) ◽  
pp. 179
Author(s):  
Kai P. Hussnaetter ◽  
Magnus Philipp ◽  
Kira Müntjes ◽  
Michael Feldbrügge ◽  
Kerstin Schipper
Keyword(s):  
Protein Production ◽  
Heterologous Protein ◽  
Ustilago Maydis ◽  
Downstream Processing ◽  
Culture Broth ◽  
Yeast Cells ◽  
Heterologous Protein Expression ◽  
Heterologous Proteins ◽  
Regulatory Systems ◽  
Unconventional Secretion

Heterologous protein production is a highly demanded biotechnological process. Secretion of the product to the culture broth is advantageous because it drastically reduces downstream processing costs. We exploit unconventional secretion for heterologous protein expression in the fungal model microorganism Ustilago maydis. Proteins of interest are fused to carrier chitinase Cts1 for export via the fragmentation zone of dividing yeast cells in a lock-type mechanism. The kinase Don3 is essential for functional assembly of the fragmentation zone and hence, for release of Cts1-fusion proteins. Here, we are first to develop regulatory systems for unconventional protein secretion using Don3 as a gatekeeper to control when export occurs. This enables uncoupling the accumulation of biomass and protein synthesis of a product of choice from its export. Regulation was successfully established at two different levels using transcriptional and post-translational induction strategies. As a proof-of-principle, we applied autoinduction based on transcriptional don3 regulation for the production and secretion of functional anti-Gfp nanobodies. The presented developments comprise tailored solutions for differentially prized products and thus constitute another important step towards a competitive protein production platform.

scholarly journals Disruption of the Complex between GAPDH and Hsp70 Sensitizes C6 Glioblastoma Cells to Hypoxic Stress

2021 ◽  
Vol 22 (4) ◽  
pp. 1520
Author(s):  
Marina A. Mikeladze ◽  
Elizaveta A. Dutysheva ◽  
Victor G. Kartsev ◽  
Boris A. Margulis ◽  
Irina V. Guzhova ◽  
...  
Keyword(s):  
Cell Death ◽  
Hypoxic Stress ◽  
C6 Cells ◽  
Tumor Resistance ◽  
Glioblastoma Cells ◽  
Enzyme Expression ◽  
Hsp70 Chaperone ◽  
Therapeutic Tools ◽  
Enzyme Molecules

Hypoxia, which commonly accompanies tumor growth, depending on its strength may cause the enhancement of tumorigenicity of cancer cells or their death. One of the proteins targeted by hypoxia is glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and we demonstrated here that hypoxia mimicked by treating C6 rat glioblastoma cells with cobalt chloride caused an up-regulation of the enzyme expression, while further elevation of hypoxic stress caused the enzyme aggregation concomitantly with cell death. Reduction or elevation of GAPDH performed with the aid of specific shRNAs resulted in the augmentation of the tumorigenicity of C6 cells or their sensitization to hypoxic stress. Another hypoxia-regulated protein, Hsp70 chaperone, was shown to prevent the aggregation of oxidized GAPDH and to reduce hypoxia-mediated cell death. In order to release the enzyme molecules from the chaperone, we employed its inhibitor, derivative of colchicine. The compound was found to substantially increase aggregation of GAPDH and to sensitize C6 cells to hypoxia both in vitro and in animals bearing tumors with distinct levels of the enzyme expression. In conclusion, blocking the chaperonic activity of Hsp70 and its interaction with GAPDH may become a promising strategy to overcome tumor resistance to multiple environmental stresses and enhance existing therapeutic tools.

Determination of Sample Size and Influencing Factors to Characterize Faulting in Jointed Concrete Pavements

2021 ◽  
pp. 036119812199670
Author(s):  
Ruohan Li ◽  
Jorge A. Prozzi
Keyword(s):  
Sample Size ◽  
Plain Concrete ◽  
Concrete Pavements ◽  
Portland Cement Concrete ◽  
Federal Highway ◽  
Significant Difference ◽  
Jointed Concrete Pavement ◽  
Functional Classes ◽  
High Precipitation ◽  
Departments Of Transportation

The objective of this study is to evaluate the field variability of jointed concrete pavement (JCP) faulting and its effects on pavement performance. The standard deviation of faulting along both the longitudinal and transverse directions are calculated. Based on these, the overall variability is determined, and the required sample sizes needed for a given precision at a certain confidence level are calculated and presented. This calculation is very important as state departments of transportation are required to report faulting every 0.1 mi to the Federal Highway Administration as required by the 2015 FAST Act. On average, twice the number of measurements are needed on jointed reinforced concrete pavements (JRCP) to achieve the same confidence and precision as on jointed plain concrete pavements (JPCP). For example, a sample size of 13 is needed to achieve a 95% confidence interval with a precision of 1.0 mm for average faulting of JPCP, while 26 measurements are required for JRCP ones. Average faulting was found to correlate with several climatic, structural, and traffic variables, while no significant difference was found between edge and outer wheelpath measurements. The application of Portland cement concrete overlay and the use of dowel bars (rather than aggregate interlock) are found to significantly reduce faulting. Older sections located on higher functional classes, and in regions of high precipitation or where the daily temperature change is larger, tend to have higher faulting, and might require larger samples sizes as compared with the rest when faulting surveys are to be conducted.

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