A novel plasmid addiction system for large-scale production of cyanophycin in Escherichia coli using mineral salts medium

2010 ◽  
Vol 89 (3) ◽  
pp. 593-604 ◽  
Author(s):  
Jens Kroll ◽  
Stefan Klinter ◽  
Alexander Steinbüchel
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Scale Production ◽  
Mineral Salts ◽  
Large Scale Production ◽  
Plasmid Addiction System

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.

scholarly journals Technical-Scale Production of Cyanophycin with Recombinant Strains of Escherichia coli

2002 ◽  
Vol 68 (7) ◽  
pp. 3377-3384 ◽  
Author(s):  
Kay M. Frey ◽  
Fred B. Oppermann-Sanio ◽  
Holger Schmidt ◽  
Alexander Steinbüchel
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Cell Mass ◽  
Extraction Procedure ◽  
Temperature Shift ◽  
Exponential Growth Phase ◽  
Acid Extraction ◽  
Scale Production ◽  
Mineral Salts ◽  
Cell Content

ABSTRACT By the use of Escherichia coli DH1 harboring cphA from Synechocystis sp. strain PCC6803, large-scale production of cyanophycin at 30- and 500-liter culture volumes was established. Transcription of cphA was controlled by the thermosensitive cI857 repressor, which enabled induction of cphA by a simple temperature shift in the culture fluid. Maximum cyanophycin cell content of up to 24% (wt/wt) of cellular dry matter was obtained by induction in the early exponential growth phase and cultivation of the cells in terrific broth complex medium. Synthesis of cyanophycin was found to be strongly dependent on the presence of complex components, and in mineral salts medium the cells synthesized and accumulated cyanophycin only if Casamino Acids were added. Cultivations were done at the 500-liter scale, allowing the provision of cell mass for the preparation of cyanophycin at the kilogram scale. Isolation of cyanophycin was achieved by a new acid extraction procedure which allowed large-scale purification of the polyamide from whole cells.

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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