Efficient production of Thermus protease aqualysin I in Escherichia coli: effects of cloned gene structure and two-stage culture

1996 ◽  
Vol 45 (1-2) ◽  
pp. 94-101 ◽  
Author(s):  
S. Sakamoto ◽  
I. Terada ◽  
Y. -C. Lee ◽  
K. Uehara ◽  
H. Matsuzawa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gene Structure ◽  
Efficient Production ◽  
Two Stage ◽  
Cloned Gene ◽  
Two Stage Culture

scholarly journals Improved Succinic Acid Production in the Anaerobic Culture of an Escherichia coli pflB ldhA Double Mutant as a Result of Enhanced Anaplerotic Activities in the Preceding Aerobic Culture

2007 ◽  
Vol 73 (24) ◽  
pp. 7837-7843 ◽  
Author(s):  
Hui Wu ◽  
Zhi-min Li ◽  
Li Zhou ◽  
Qin Ye
Keyword(s):  
Escherichia Coli ◽  
Succinic Acid ◽  
Acid Production ◽  
Double Mutant ◽  
Anaerobic Culture ◽  
Two Stage ◽  
Acid Yield ◽  
Succinic Acid Production ◽  
Ferment Glucose ◽  
Two Stage Culture

ABSTRACT Escherichia coli NZN111 is a pflB ldhA double mutant which loses its ability to ferment glucose anaerobically due to redox imbalance. In this study, two-stage culture of NZN111 was carried out for succinic acid production. It was found that when NZN111 was aerobically cultured on acetate, it regained the ability to ferment glucose with succinic acid as the major product in subsequent anaerobic culture. In two-stage culture carried out in flasks, succinic acid was produced at a level of 11.26 g/liter from 13.4 g/liter of glucose with a succinic acid yield of 1.28 mol/mol glucose and a productivity of 1.13 g/liter·h in the anaerobic stage. Analyses of key enzyme activities revealed that the activities of isocitrate lyase, malate dehydrogenase, malic enzyme, and phosphoenolpyruvate (PEP) carboxykinase were greatly enhanced while those of pyruvate kinase and PEP carboxylase were reduced in the acetate-grown cells. The two-stage culture was also performed in a 5-liter fermentor without separating the acetate-grown NZN111 cells from spent medium. The overall yield and concentration of succinic acid reached 1.13 mol/mol glucose and 28.2 g/liter, respectively, but the productivity of succinic acid in the anaerobic stage dropped to 0.7 g/liter·h due to cell autolysis and reduced anaplerotic activities. The results indicate the great potential to take advantage of cellular regulation mechanisms for improvement of succinic acid production by a metabolically engineered E. coli strain.

Hybrid two-stage culture of Halamphora coffeaeformis for biodiesel production: Growth phases, nutritional stages and biorefinery approach

2018 ◽  
Vol 118 ◽  
pp. 984-992 ◽  
Author(s):  
Lucas A. Martín ◽  
Cecilia A. Popovich ◽  
Ana M. Martínez ◽  
Paola G. Scodelaro Bilbao ◽  
María C. Damiani ◽  
...  
Keyword(s):  
Biodiesel Production ◽  
Growth Phases ◽  
Two Stage ◽  
Production Growth ◽  
Two Stage Culture

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.

Optimized Culture Conditions for the Efficient Production of Porcine Adenylate Kinase in Recombinant Escherichia coli

2010 ◽  
Vol 162 (3) ◽  
pp. 823-829 ◽  
Author(s):  
Toru Matsui ◽  
Takashi Togari ◽  
Satoru Misawa ◽  
Tomoyuki Namihira ◽  
Naoya Shinzato ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Adenylate Kinase ◽  
Culture Conditions ◽  
Recombinant Escherichia Coli ◽  
Efficient Production

Engineering efficient production of itaconic acid from diverse substrates in Escherichia coli

2017 ◽  
Vol 249 ◽  
pp. 73-81 ◽  
Author(s):  
Peiching Chang ◽  
Grey S. Chen ◽  
Hsiang-Yuan Chu ◽  
Ken W. Lu ◽  
Claire R. Shen
Keyword(s):  
Escherichia Coli ◽  
Itaconic Acid ◽  
Efficient Production

scholarly journals Metabolic engineering of Escherichia coli W for isobutanol production on chemically defined medium and cheese whey as alternative raw material

2020 ◽  
Vol 47 (12) ◽  
pp. 1117-1132
Author(s):  
Katharina Novak ◽  
Juliane Baar ◽  
Philipp Freitag ◽  
Stefan Pflügl
Keyword(s):  
Escherichia Coli ◽  
Cheese Whey ◽  
Strain Improvement ◽  
Defined Medium ◽  
Raw Material ◽  
Efficient Production ◽  
Substrate Uptake ◽  
Chemically Defined Medium ◽  
E Coli ◽  
Plasmid Library

AbstractThe aim of this study was to establish isobutanol production on chemically defined medium in Escherichia coli. By individually expressing each gene of the pathway, we constructed a plasmid library for isobutanol production. Strain screening on chemically defined medium showed successful production in the robust E. coli W strain, and expression vector IB 4 was selected as the most promising construct due to its high isobutanol yields and efficient substrate uptake. The investigation of different aeration strategies in combination with strain improvement and the implementation of a pulsed fed-batch were key for the development of an efficient production process. E. coli W ΔldhA ΔadhE Δpta ΔfrdA enabled aerobic isobutanol production at 38% of the theoretical maximum. Use of cheese whey as raw material resulted in longer process stability, which allowed production of 20 g l−1 isobutanol. Demonstrating isobutanol production on both chemically defined medium and a residual waste stream, this study provides valuable information for further development of industrially relevant isobutanol production processes.

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