scholarly journals A Novel Method for High-Level Production of TEV Protease by Superfolder GFP Tag

2009 ◽  
Vol 2009 ◽  
pp. 1-8 ◽  
Author(s):  
Xudong Wu ◽  
Di Wu ◽  
Zhisheng Lu ◽  
Wentao Chen ◽  
Xiaojian Hu ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Large Scale ◽  
Fluorescent Protein ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Scale Production ◽  
Sequence Specificity ◽  
Tev Protease ◽  
Large Scale Production ◽  
Novel Method

Because of its stringent sequence specificity, tobacco etch virus (TEV) protease is widely used to remove fusion tags from recombinant proteins. Due to the poor solubility of TEV protease, many strategies have been employed to increase the expression level of this enzyme. In our work, we introduced a novel method to produce TEV protease by using visible superfolder green fluorescent protein (sfGFP) as the fusion tag. The soluble production and catalytic activity of six variants ofsfGFP-TEV was examined, and then the best variant was selected for large-scale production. After purified by Ni-NTA affinity chromatography and Q anion exchange chromatography, the best variant ofsfGFP-TEV fusion protease was obtained with purity of over 98% and yield of over 320 mg per liter culture. ThesfGFP-TEV had a similar catalytic activity to that of the original TEV protease. Our research showed a novel method of large-scale production of visible and functional TEV protease for structural genomics research and other applications.

A Novel Method for Expression and Large-Scale Production of Human Brain -Glutamate Decarboxylase

2000 ◽  
Vol 267 (3) ◽  
pp. 777-782 ◽  
Author(s):  
Kathleen M. Davis ◽  
Todd Foos ◽  
C.Sean Bates ◽  
Erin Tucker ◽  
Che-Chang Hsu ◽  
...  
Keyword(s):  
Human Brain ◽  
Glutamate Decarboxylase ◽  
Large Scale ◽  
Scale Production ◽  
Large Scale Production ◽  
Novel Method

scholarly journals Large-scale Production of Novel Porcine Circovirus Type 2d (PCV2d) Subunit Vaccine Using E. coli Platform

2021 ◽  
Author(s):  
Sarawuth Noppiboon ◽  
Thunyaluk Kerdboon ◽  
Neeracha Lapanusorn ◽  
Pisit Ekkpongpaisit ◽  
Sarah Slack ◽  
...  
Keyword(s):  
Large Scale ◽  
Pilot Scale ◽  
Porcine Circovirus Type ◽  
Exchange Chromatography ◽  
Porcine Circovirus ◽  
Scale Production ◽  
E Coli ◽  
Large Scale Production ◽  
Production Platform ◽  
Cost Efficient

Porcine Circovirus type 2d (PCV2d) is becoming the predominant PCV genotype and considerably affects the global pig industry. Nevertheless, currently no commercial PCV2d vaccine is available. Preventing and controlling the disease caused by PCV2d is therefore based on other genotype-based vaccines. However, their production platforms are laborious, limited in expression level and relatively expensive for veterinary applications. To address these challenges, we have developed a simple and cost-efficient platform for a novel PCV2d vaccine production platform, using fed-batch E. coli fermentation followed by cell disruption and filtration, and a single purification step via cation exchange chromatography. The process was developed at bench scale and then pilot scale where the PCV2d subunit protein yield was approximately 0.8 g/L fermentation volume in a short production time. Moreover, we have successfully implemented this production process at two different sites, in Southeast Asia and Europe. This demonstrates transferability and the high potential for successful industrial production.

scholarly journals Antioxidant activity of pea protein hydrolysates produced by batch fermentation with lactic acid bacteria

2015 ◽  
Vol 67 (3) ◽  
pp. 1033-1042 ◽  
Author(s):  
Nemanja Stanisavljevic ◽  
Goran Vukotic ◽  
Ferenc Pastor ◽  
Desanka Suznjevic ◽  
Zivko Jovanovic ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Large Scale ◽  
Lactobacillus Rhamnosus ◽  
Weight Fraction ◽  
Exchange Chromatography ◽  
Scale Production ◽  
Large Scale Production ◽  
Pea Proteins ◽  
Basic Peptides ◽  
Pea Seed

Nine Lactobacillus strains known for surface proteinase activity were chosen from our collection and tested for their ability to grow in pea seed protein-based medium, and to hydrolyze purified pea proteins in order to produce peptides with antioxidant (AO) activity. Two strains, Lactobacillus rhamnosus BGT10 and Lactobacillus zeae LMG17315, exhibited strong proteolytic activity against pea proteins. The AO activity of the pea hydrolysate fraction, MW <10 kDa, obtained by the fermentation of purified pea proteins with Lactobacillus rhamnosus BGT10, was tested by standard spectrophotometric assays (DPPH, ABTS, Fe3+-reducing capacity) and the recently developed direct current (DC) polarographic assay. The low molecular weight fraction of the obtained hydrolysate was separated using ion exchange chromatography, while the AO activity of eluted fractions was determined by means of a sensitive DC polarographic assay without previous concentration of samples. Results revealed that the fraction present in low abundance that contained basic peptides possessed the highest antioxidant activity. Based on the obtained results, it can be concluded that Lactobacillus rhamnosus BGT10 should be further investigated as a candidate strain for large-scale production of bioactive peptides from legume proteins.

Large-scale production of high-quality graphene using glucose and ferric chloride

2014 ◽  
Vol 5 (12) ◽  
pp. 4656-4660 ◽  
Author(s):  
Binbin Zhang ◽  
Jinliang Song ◽  
Guanying Yang ◽  
Buxing Han
Keyword(s):  
Ferric Chloride ◽  
Large Scale ◽  
Scale Production ◽  
High Quality ◽  
Large Scale Production ◽  
Novel Method

A novel method to prepare high-quality graphene is developed using simple calcination of a mixture of glucose and FeCl3.

A novel method for the large-scale production of PG-CNP37, a C-type natriuretic peptide analogue

2013 ◽  
Vol 164 (2) ◽  
pp. 196-201 ◽  
Author(s):  
Shinong Long ◽  
Daniel J. Wendt ◽  
Sean M. Bell ◽  
Timothy W. Taylor ◽  
Jean-Yves Dewavrin ◽  
...  
Keyword(s):  
Natriuretic Peptide ◽  
Large Scale ◽  
Scale Production ◽  
Peptide Analogue ◽  
Large Scale Production ◽  
Novel Method

scholarly journals Production and Partial Purification of Heat-Stable Enterotoxin (A) Produced by Enterotoxigenic Escherichia coli

2010 ◽  
Vol 34 (1) ◽  
pp. 122-129
Author(s):  
Rajiha I. Al-Nuaimy
Keyword(s):  
Large Scale ◽  
Specific Activity ◽  
Exchange Chromatography ◽  
Enterotoxigenic Escherichia Coli ◽  
Partial Purification ◽  
Scale Production ◽  
E Coli ◽  
Standard Curve ◽  
Heat Stable ◽  
Large Scale Production

A total of (25) stool samples were collected from children and adults (2- 4) years oldsuffering from diarrhea to isolate E. coli strains that produce heat-stable enterotoxin a (STa),and after performing microscopic examination, cultural characterization and biochemicalidentification only (11) isolates showed positive E. coli. STa activity was estimated by usingsuckling mouse assay (SMA) and from these (11) isolates only (5) showed STa activity andthe one with the highest STa activity was selected for large scale production of STa, whichwas followed by partial purification using ion-exchange chromatography (normal phase)using DEAE sephadex A-50 column. After purification and determination of proteinconcentration by using the standard curve of bovine serum albumin, the concentration oftoxin-protein was estimated as (1.08) mg/ml. The specific activity varied from (350) U/mgprotein at the first step of purification to (2366.6) U/mg protein at the final step, while thefinal purification of the toxin was about (6.76) fold and with a yield of (18.25) %.

I. S. A. Baud. Forms of Production and Women's Labour: Gender Aspects of Industrialisation in India and Mexico. New Delhi: Sage Publications. 1992. 321 pp.Hardbound. Price: Indian Rs 295.00.

1993 ◽  
Vol 32 (1) ◽  
pp. 129-131
Author(s):  
Naureen Talha
Keyword(s):  
Large Scale ◽  
Indian Society ◽  
New Delhi ◽  
Scale Production ◽  
Female Labour ◽  
Self Employment ◽  
Commodity Production ◽  
Large Scale Production ◽  
Mexican Society ◽  
Small Production

The literature on female labour in Third World countries has become quite extensive. India, being comparatively more advanced industrially, and in view of its size and population, presents a pictures of multiplicity of problems which face the female labour market. However, the author has also included Mexico in this analytical study. It is interesting to see the characteristics of developing industrialisation in two different societies: the Indian society, which is conservative, and the Mexican society, which is progressive. In the first chapter of the book, the author explains that he is not concerned with the process of industrialisation and female labour employed at different levels of work, but that he is interested in forms of production and women's employment in large-scale production, petty commodity production, marginal small production, and self-employment in the informal sector. It is only by analysis of these forms that the picture of females having a lower status is understood in its social and political setting.

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