Large-scale production and characterization of Bacillus thuringiensis subsp. tenebrionis insecticidal protein from Escherichia coli

1997 ◽  
Vol 47 (3) ◽  
pp. 255-261 ◽  
Author(s):  
M. E. Gustafson ◽  
R. A. Clayton ◽  
P. B. Lavrik ◽  
G. V. Johnson ◽  
R. M. Leimgruber ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacillus Thuringiensis ◽  
Large Scale ◽  
Insecticidal Protein ◽  
Scale Production ◽  
Bacillus Thuringiensis Subsp ◽  
Large Scale Production

Characterization of Di-Rhamnolipid Biosurfactant in Recombinant Escherichia coli

2021 ◽  
Vol 874 ◽  
pp. 107-114
Author(s):  
Subhan Hadi Kusuma ◽  
Karlia Meitha ◽  
Sony Suhandono
Keyword(s):  
Escherichia Coli ◽  
Large Scale ◽  
Sodium Dodecyl ◽  
Petroleum Industry ◽  
Scale Production ◽  
Glycolipid Biosurfactant ◽  
Promoter Characterization ◽  
Large Scale Production ◽  
Rhamnolipid Biosurfactant

Surfactants are amphiphilic molecules, which have hydrophilic and hydrophobic groups. Surfactants have an important role in various fields including agriculture, cosmetics, pharmaceuticals, bioremediation, and the petroleum industry especially EOR but because synthetic surfactants are not biodegradable, it is necessary to produce biodegradable surfactants such as rhamnolipid biosurfactants. Rhamnolipid is a glycolipid biosurfactant produced by Pseudomonas aeuruginosa. This species is a pathogen, so it is needed to overcome this problem by cloning the rhamnolipid gene into Escherichia coli for large-scale production. Rhamnolipid biosynthesis includes three main genes, rhlA, rhlB, and rhlC. The rhlAB produces mono-rhamnolipid and rhlABC produces di-rhamnolipid. The construction involved one plasmid pPM RHLABC (di-rhamnolipid) with T7lac promoter. Characterization of surfactants by E24, IFT, and CMC analysis showed that di-rhamnolipid biosurfactant has the best activity (70%, 0.8 mN/m, and 300 mg/L) than chemical surfactant, sodium dodecyl sulfate (46%, 4.7 mN/m, and 2000 mg/L) at pH 7, 25 °C, and 0% salinity. The conclusion from this research shows that the characteristics of di-rhamnolipid are very promising in the utilization of industrial-scale including EOR technology, agriculture, and pharmacy

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 Large-scale production and physicochemical characterization of human immune interferon.

1979 ◽  
Vol 26 (1) ◽  
pp. 36-41 ◽  
Author(s):  
M P Langford ◽  
J A Georgiades ◽  
G J Stanton ◽  
F Dianzani ◽  
H M Johnson
Keyword(s):  
Large Scale ◽  
Physicochemical Characterization ◽  
Scale Production ◽  
Immune Interferon ◽  
Large Scale Production ◽  
Human Immune

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 Preclinical Characterization of Naturally Occurring Polyketide Cyclophilin Inhibitors from the Sanglifehrin Family

2011 ◽  
Vol 55 (5) ◽  
pp. 1975-1981 ◽  
Author(s):  
Matthew A. Gregory ◽  
Michael Bobardt ◽  
Susan Obeid ◽  
Udayan Chatterji ◽  
Nigel J. Coates ◽  
...  
Keyword(s):  
Large Scale ◽  
Scale Production ◽  
Subgenomic Replicon ◽  
Isomerase Activity ◽  
Lead Compounds ◽  
Naturally Occurring ◽  
Large Scale Production ◽  
Pharmacokinetic Properties ◽  
Biosynthetic Engineering

ABSTRACTCyclophilin inhibitors currently in clinical trials for hepatitis C virus (HCV) are all analogues of cyclosporine (CsA). Sanglifehrins are a group of naturally occurring cyclophilin binding polyketides that are structurally distinct from the cyclosporines and are produced by a microorganism amenable to biosynthetic engineering for lead optimization and large-scale production by fermentation. Preclinical characterization of the potential utility of this class of compounds for the treatment of HCV revealed that the natural sanglifehrins A to D are all more potent than CsA at disrupting formation of the NS5A-CypA, -CypB, and -CypD complexes and at inhibition of CypA, CypB, and CypD isomerase activity. In particular, sanglifehrin B (SfB) was 30- to 50-fold more potent at inhibiting the isomerase activity of all Cyps tested than CsA and was also shown to be a more potent inhibitor of the 1b subgenomic replicon (50% effective concentrations [EC50s] of 0.070 μM and 0.16 μM in Huh 5-2 and Huh 9-13 cells, respectively). Physicochemical and mouse pharmacokinetic analyses revealed low oral bioavailability (F< 4%) and low solubility (<25 μM), although the half-lives (t1/2) of SfA and SfB in mouse blood after intravenous (i.v.) dosing were long (t1/2> 5 h). These data demonstrate that naturally occurring sanglifehrins are suitable lead compounds for the development of novel analogues that are less immunosuppressive and that have improved metabolism and pharmacokinetic properties.

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