Engineering the precursor pool to modulate the production of pamamycins in the heterologous host S. albus J1074

2021 ◽  
Author(s):  
Nils Gummerlich ◽  
Niko Manderscheid ◽  
Yuriy Rebets ◽  
Maksym Myronovskyi ◽  
Lars Gläser ◽  
...  
Keyword(s):  
Heterologous Host ◽  
Precursor Pool

Agrobacteium Transformation of Tobacco with a Genetic Module of Antimalarial Agent Artemisinin Biosynthesis

2020 ◽  
Vol 36 (3) ◽  
pp. 34-45
Author(s):  
T.Yu. Mitiuchkina ◽  
A.S. Pushin ◽  
A.K. Tzareva ◽  
A.M. Vainstein ◽  
S.V. Dolgov
Keyword(s):  
Target Genes ◽  
Artemisia Annua ◽  
Biosynthesis Pathway ◽  
Expression Systems ◽  
Heterologous Host ◽  
Russian Science ◽  
Tobacco Plants ◽  
Genetic Module ◽  
Heterologous Expression Systems ◽  
Artemisinin Biosynthesis

Artemisinin-based medicines are the most effective treatment for malaria. To date, the wormwood plants (Artemisia annua L.) are the main source of artemisinin. Due to the limited nature of this source, considerable efforts are directed towards the development of methods for artemisinin production via heterologous expression systems. We used in this study agrobacterial transformation to transfer the genetic module of the artemisinin biosynthesis pathway into plants and then analyzed its transcription in a heterologous host. Tobacco plants were transformed with the artemisinin biosynthesis genes encoding amorpha-4,11-diene synthase, artemisin-aldehyde All(13) reductase, amorpha-4,11-diene monooxygenase, cytochrome P450 reductase from A. annua and yeast 3-hydroxy-3-methylglutaryl-coenzyme A reductase cloned in the pArtemC vector; farnesyl diphosphate synthase and aldehyde dehydrogenase were used to transform the plants as parts of vector p2356. As a result of transformation with the pArtemC and p2356 vectors, in twos transgenic lines with all target genes were obtained. Five genes of artemisinin biosynthesis and two genes of biosynthesis of its precursors were successfully transferred into the genome of transgenic tobacco lines as a result of the co-transformation with abovementioned vectors. Thus, the entire artemisinin biosynthesis pathway was first reconstructed in heterologous plants: the transcription of the artemisinin biosynthesis genes in the tobacco plants was shown via RT-PCR. The obtained results will be used in further research on expression systems for the production of artemisinin and other non-protein substances in heterologous host plants. artemisinin, malaria, metabolic engineering, tobacco, transgenic plants This work was supported by a Grant from the Russian Science Foundation no. 19-14-00190.

scholarly journals Total Biosynthesis and Diverse Applications of the Nonribosomal Peptide-Polyketide Siderophore Yersiniabactin

2015 ◽  
Vol 81 (16) ◽  
pp. 5290-5298 ◽  
Author(s):  
Mahmoud Kamal Ahmadi ◽  
Samar Fawaz ◽  
Charles H. Jones ◽  
Guojian Zhang ◽  
Blaine A. Pfeifer
Keyword(s):  
Polyketide Synthase ◽  
Parallel Applications ◽  
Total Removal ◽  
Heterologous Host ◽  
Nonribosomal Peptide ◽  
Content Type ◽  
Metal Chelating ◽  
Removal Capacity ◽  
Peptide Synthetase ◽  
Host Infection

ABSTRACTYersiniabactin (Ybt) is a mixed nonribosomal peptide-polyketide natural product natively produced by the pathogenYersinia pestis. The compound enables iron scavenging capabilities upon host infection and is biosynthesized by a nonribosomal peptide synthetase featuring a polyketide synthase module. This pathway has been engineered for expression and biosynthesis usingEscherichia colias a heterologous host. In the current work, the biosynthetic process for Ybt formation was improved through the incorporation of a dedicated step to eliminate the need for exogenous salicylate provision. When this improvement was made, the compound was tested in parallel applications that highlight the metal-chelating nature of the compound. In the first application, Ybt was assessed as a rust remover, demonstrating a capacity of ∼40% compared to a commercial removal agent and ∼20% relative to total removal capacity. The second application tested Ybt in removing copper from a variety of nonbiological and biological solution mixtures. Success across a variety of media indicates potential utility in diverse scenarios that include environmental and biomedical settings.

Water-soluble phospholipid precursor pool-sizes in quick-frozen and unfrozen rat livers

1975 ◽  
Vol 69 (1) ◽  
pp. 300-305 ◽  
Author(s):  
Elizabeth K. Korniat ◽  
Donald A. Beeler
Keyword(s):  
Water Soluble ◽  
Precursor Pool ◽  
Rat Livers ◽  
Pool Sizes

Monorhamnolipids and 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs) production using Escherichia coli as a heterologous host

2006 ◽  
Vol 73 (1) ◽  
pp. 187-194 ◽  
Author(s):  
Natividad Cabrera-Valladares ◽  
Anne-Pascale Richardson ◽  
Clarita Olvera ◽  
Luis Gerardo Treviño ◽  
Eric Déziel ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Heterologous Host ◽  
Alkanoic Acids

scholarly journals Erythropoietin-Induced Erythroid Precursor Pool Depletion Causes Erythropoietin Hyporesponsiveness

2012 ◽  
Vol 30 (4) ◽  
pp. 1026-1036
Author(s):  
Xiaoyu Yan ◽  
Sihem Ait-Oudhia ◽  
Wojciech Krzyzanski
Keyword(s):  
Erythroid Precursor ◽  
Precursor Pool

THE REDUCED CAPACITY OF THROMBOXANE FORMATION IN THROMBIN-PREACTIVATED PLATELETS IS NOT CAUSED BY DEPLETION OF THEIR ARACHIDONIC ACID POOL

1987 ◽  
Author(s):  
R E Scharf ◽  
M Stockschläder ◽  
H J Reimers ◽  
W Schneider
Keyword(s):  
Arachidonic Acid ◽  
Human Platelets ◽  
Thrombin Receptor ◽  
Precursor Pool ◽  
Acid Pool ◽  
Exogenous Arachidonic Acid ◽  
High Concentrations ◽  
Short Time ◽  
Thromboxane Formation

Thromboxane (TX) synthesis of washed human platelets pretreated with high concentrations of thrombin (0.5-2.0 U/ml) for 20 sec is significantly reduced upon further thrombin stimulation. Compared to controls (tyrode-pretreated platelets), thrombin-preactivated platelets recover normal TX synthesis following exposure to exogenous arachidonic acid (AA) indicating that short-time thrombin treatment does not inactivate platelet cyclooxygenase or TX synthetase (Blood 63: 858, 1984). To evaluate whether the reduced TX synthesis upon -the second thrombin exposure is due to depletion of their AA precursor pool, thrombin-pretreated platelets and tyrode-pretreated platelets (5×108/ml) were resuspended in autologous ACD plasma and incubated at 37°C with 0.2 μCi 14C-AA (20 μM) for 60 to 90 min in the presence of PGE1 (10 μM). Mean platelet uptake of 14C-AA (disappearance of radioactivity from the supernatant) was 12+3 nmoles AA/109 platelets and did not differ significantly between thrombin-pretreated platelets and controls. Thrombin-pretreated platelets released 10% or 4.5% of their 14c-activity upon further exposure to thrombin (2 U/ml) or collagen (8 μg/ml), respectively. The release from control platelets (15% with thrombin, 6.5% with collagen) did not differ from that of thrombin-pretreated platelets. However, even after incubation in ACD plasma, thrombin-pretreated platelets continued to form significantly less TXB2 (5.0±1.6 nmoles/109 platelets) than controls (9.7±2.2 nmoles/109 platelets, p< 0.05). These data indicate that the reduced capacity of thrombin-pretreated platelets is due neither to a depletion of the endogenous AA pool nor to an inactivation of cyclooxygenase or TX synthetase. The reduced TX synthesis capacity may be caused by a modification, destruction or desensitization of the platelet thrombin receptor as a consequence of the preceding thrombin stimulation.

Glutamine prevents downregulation of myosin heavy chain synthesis and muscle atrophy from glucocorticoids

1995 ◽  
Vol 268 (4) ◽  
pp. E730-E734 ◽  
Author(s):  
R. C. Hickson ◽  
S. M. Czerwinski ◽  
L. E. Wegrzyn
Keyword(s):  
Protein Synthesis ◽  
Myosin Heavy Chain ◽  
Muscle Mass ◽  
Muscle Atrophy ◽  
Heavy Chain ◽  
Total Protein ◽  
Female Rats ◽  
Glucocorticoid Treatment ◽  
Precursor Pool ◽  
Chain Synthesis

The aims of this study were to determine whether glutamine infusion prevents the decline in protein synthesis and muscle wasting associated with repeated glucocorticoid treatment. Hormone (cortisol acetate, 100 mg.kg body wt-1.day-1) and vehicle (carboxymethyl cellulose)-treated female rats were infused with either saline or glutamine (240 mM, 0.75 ml/h) for a 7-day period. Glutamine infusion attenuated the decline of plantaris muscle glutamine concentration (3.0 +/- 0.2 vs. 2.3 +/- 0.2 mumol/g) and prevented > 70% of the total muscle mass losses due to the glucocorticoid injections. Fractional synthesis rates of myosin heavy chain (MHC) and total protein were determined after constant [3H]leucine infusion from the leucyl-tRNA precursor pool, which was similar in all groups (range 4.8 +/- 0.5 to 6.3 +/- 0.4 disintegrations.min-1.pmol-1). MHC synthesis rates (%/day) in plantaris muscles were reduced to approximately 40% of controls (4.2/9.4). Although glutamine had no effect on MHC synthesis in vehicle-treated animals (10.1/9.4), it prevented 50% (7.6/4.2) of the hormone-induced decline in MHC synthesis rates. The same results were obtained with total protein synthesis measurements. Changes in muscle mass did not appear related to estimates of protein breakdown. In conclusion, these data show that glutamine infusion is effective therapy in counteracting glucocorticoid-induced muscle atrophy. Atrophy attenuation appears related to maintaining muscle glutamine levels, which in turn may limit the glucocorticoid-mediated downregulation of MHC synthesis.

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