Process optimization for large-scale production of TGF- ? -PE40 in recombinant Escherichia coli : effect of medium composition and induction timing on protein expression

1997 ◽  
Vol 18 (4) ◽  
pp. 260-266 ◽  
Author(s):  
C Lee ◽  
W-J Sun ◽  
B W Burgess ◽  
B H Junker ◽  
J Reddy ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Expression ◽  
Process Optimization ◽  
Large Scale ◽  
Medium Composition ◽  
Recombinant Escherichia Coli ◽  
Scale Production ◽  
Large Scale Production

Large-scale production of citrate synthase from a cloned gene

1982 ◽  
Vol 60 (12) ◽  
pp. 1143-1147 ◽  
Author(s):  
Harry W. Duckworth ◽  
Alexander W. Bell
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Citrate Synthase ◽  
Specific Activity ◽  
Tetracycline Resistance ◽  
Scale Production ◽  
Ampicillin Resistance ◽  
Colicin E1 ◽  
Large Scale Production ◽  
Cloned Gene

Starting with a colicin E1 resistance recombinant plasmid which contains gltA, the gene for citrate synthase in Escherichia coli, we have constructed an ampicillin-resistance plasmid containing the gltA region as a 2.9-kilobase-pair insert in the tetracycline-resistance region of pBR322. Escherichia coli HB101 harbouring this plasmid, when grown on rich medium containing ampicillin, contains citrate synthase as about 8% of its soluble protein. The enzyme has been purified from this rich source and is identical to the chromosomal enzyme prepared previously in every property tested, except for specific activity, which is 64 U∙mg−1 as compared with 45–50 U∙mg−1 previously obtained. The N-terminal sequences of both enzymes are reported, and they are identical up to residue 16 at least. The overall yield of pure enzyme, starting with the cells grown in 15 L of medium, is 600–800 mg.

scholarly journals Efficient Production of l-Ribose with a Recombinant Escherichia coli Biocatalyst

2008 ◽  
Vol 74 (10) ◽  
pp. 2967-2975 ◽  
Author(s):  
Ryan D. Woodyer ◽  
Nathan J. Wymer ◽  
F. Michael Racine ◽  
Shama N. Khan ◽  
Badal C. Saha
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Active Form ◽  
Apium Graveolens ◽  
Efficient Production ◽  
Scale Production ◽  
Gene Encoding ◽  
Large Scale Production ◽  
One Step ◽  
Rare Sugars

ABSTRACT A new synthetic platform with potential for the production of several rare sugars, with l-ribose as the model target, is described. The gene encoding the unique NAD-dependent mannitol-1-dehydrogenase (MDH) from Apium graveolens (garden celery) was synthetically constructed for optimal expression in Escherichia coli. This MDH enzyme catalyzes the interconversion of several polyols and their l-sugar counterparts, including the conversion of ribitol to l-ribose. Expression of recombinant MDH in the active form was successfully achieved, and one-step purification was demonstrated. Using the created recombinant E. coli strain as a whole-cell catalyst, the synthetic utility was demonstrated for production of l-ribose, and the system was improved using shaken flask experiments. It was determined that addition of 50 to 500 μM ZnCl2 and addition of 5 g/liter glycerol both improved production. The final levels of conversion achieved were >70% at a concentration of 40 g/liter and >50% at a concentration of 100 g/liter. The best conditions determined were then scaled up to a 1-liter fermentation that resulted in 55% conversion of 100 g/liter ribitol in 72 h, for a volumetric productivity of 17.4 g liter−1 day−1. This system represents a significantly improved method for the large-scale production of l-ribose.

pHsh vectors, a novel expression system of Escherichia coli for the large-scale production of recombinant enzymes

2010 ◽  
Vol 32 (6) ◽  
pp. 795-801 ◽  
Author(s):  
Huawei Wu ◽  
Jianjun Pei ◽  
Yu Jiang ◽  
Xin Song ◽  
Weilan Shao
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Expression System ◽  
Scale Production ◽  
Recombinant Enzymes ◽  
Large Scale Production

Large-Scale Production of Major House Dust Mite Allergen Der f 2 Mutant (C8/119S) in Escherichia coli

2008 ◽  
Vol 106 (4) ◽  
pp. 387-392 ◽  
Author(s):  
Satoshi Koyanagi ◽  
Toshihiro Maeda ◽  
Toshio Murakami ◽  
Kenjirou Kawatsu ◽  
Keishin Sugawara ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
House Dust ◽  
House Dust Mite ◽  
Large Scale ◽  
Mite Allergen ◽  
Scale Production ◽  
House Dust Mite Allergen ◽  
Large Scale Production ◽  
Dust Mite ◽  
Dust Mite Allergen
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