Cloning and expression of phosphoenolpyruvate carboxykinase from a cestode parasite and its solubilization from inclusion bodies using l-arginine

Dutta, Asim Kumar; Ramnath,; Dkhar, Barilin; Tandon, Veena; Das, Bidyadhar

doi:10.1016/j.pep.2015.09.004

Cited by 3 publications

(4 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, in some cases inclusion bodies were demonstrated as retaining their protein function, suggesting functional folding to have occurred. This has been demonstrated in the context of enzymes, such as β-galactosidase, anionic tobacco peroxidase, phosphoenolpyruvate carboxykinase and pyruvate oxidase, [56,77,78,79,80,81]. Importantly, these inclusion bodies still exhibited their corresponding enzymatic activities, thereby suggesting that they contain significant amounts of properly folded proteins [77,78,80,81].…”

Section: Intracellular Self-assembly Towards In Situ Enzyme Immobimentioning

confidence: 93%

“…Thus, the enzyme activity exhibited by inclusion bodies not only demonstrates the incorporation of functional enzyme into their structures but also emphasizes the potential applications of enzyme inclusion bodies as biologically active catalyst platforms. Self-assembly-based enzyme inclusion bodies represent a novel technique for carrier-free immobilization of biologically active enzyme, and they have been demonstrated as reusable and stable across a range of conditions, which makes them suitable for potential applications in food production and biotechnological industries [49,60,79,81].…”

Section: Intracellular Self-assembly Towards In Situ Enzyme Immobimentioning

confidence: 99%

See 1 more Smart Citation

Enzyme Engineering for In Situ Immobilization

2016

View full text Add to dashboard Cite

Enzymes are used as biocatalysts in a vast range of industrial applications. Immobilization of enzymes to solid supports or their self-assembly into insoluble particles enhances their applicability by strongly improving properties such as stability in changing environments, re-usability and applicability in continuous biocatalytic processes. The possibility of co-immobilizing various functionally related enzymes involved in multistep synthesis, conversion or degradation reactions enables the design of multifunctional biocatalyst with enhanced performance compared to their soluble counterparts. This review provides a brief overview of up-to-date in vitro immobilization strategies while focusing on recent advances in enzyme engineering towards in situ self-assembly into insoluble particles. In situ self-assembly approaches include the bioengineering of bacteria to abundantly form enzymatically active inclusion bodies such as enzyme inclusions or enzyme-coated polyhydroxyalkanoate granules. These one-step production strategies for immobilized enzymes avoid prefabrication of the carrier as well as chemical cross-linking or attachment to a support material while the controlled oriented display strongly enhances the fraction of accessible catalytic sites and hence functional enzymes.

show abstract

Section: Intracellular Self-assembly Towards In Situ Enzyme Immobimentioning

confidence: 93%

Section: Intracellular Self-assembly Towards In Situ Enzyme Immobimentioning

confidence: 99%

Enzyme Engineering for In Situ Immobilization

2016

View full text Add to dashboard Cite

show abstract

“…The effect of the phytoestrogens on conformation of the purified recombinant proteins was analysed with and without the phytoestrogens using CD spectrophotometer (JASCO-815) following the method described by Dutta et al (2016 b ).…”

Section: Methodsmentioning

confidence: 99%

“…For host PEPCK, the available crystal structure of chicken mitochondrial PEPCK structure (PDB ID 2QZY) was selected. However, for parasite PEPCK, the homology model described by Dutta et al (2016 b ) was used.…”

Section: Methodsmentioning

confidence: 99%

Biological significance of phosphoenolpyruvate carboxykinase in a cestode parasite,Raillietina echinobothridaand effect of phytoestrogens on the enzyme from the parasite and its host,Gallus domesticus

et al. 2017

Self Cite

View full text Add to dashboard Cite

Phosphoenolpyruvate carboxykinase (PEPCK) is involved in glycolysis in the cestode parasite, Raillietina echinobothrida; whereas, it executes a gluconeogenic role in its host, Gallus domesticus. Because of its differing primary function in the cestode parasite and its host, this enzyme is regarded as a plausible anthelmintic target. Hence, the biological significance of PEPCK in the parasite was analysed using siRNA against PEPCK from R. echinobothrida (RePEPCK). In order to find out the functional differences between RePEPCK and GdPEPCK (PEPCK from its host, G. domesticus), PEPCK genes from both sources were cloned, over-expressed, characterized, and some properties of the purified enzymes were compared. RePEPCK and GdPEPCK showed a standard Michaelis-Menten kinetics with K mapp of 46.9 and 22.9 µ m, respectively, for phosphoenolpyruvate and K mapp of 15.4 µ m for oxaloacetate in GdPEPCK decarboxylation reaction. Here, we report antagonist behaviours of recombinant PEPCKs derived from the parasite and its host. In search of possible modulators for PEPCK, few phytoestrogens were examined on the purified enzymes and their inhibitory constants were determined and discussed. This study stresses the potential of these findings to validate PEPCK as the anthelmintic drug target for parasitism management.

show abstract

Purification and characterization of human glycerol 3-phosphate dehydrogenases (mitochondrial and cytosolic) by NAD+/NADH redox method

Syngkli

Das

2023

Biochimie

View full text Add to dashboard Cite

Cloning and expression of phosphoenolpyruvate carboxykinase from a cestode parasite and its solubilization from inclusion bodies using l-arginine

Cited by 3 publications

References 45 publications

Enzyme Engineering for In Situ Immobilization

Enzyme Engineering for In Situ Immobilization

Biological significance of phosphoenolpyruvate carboxykinase in a cestode parasite,Raillietina echinobothridaand effect of phytoestrogens on the enzyme from the parasite and its host,Gallus domesticus

Purification and characterization of human glycerol 3-phosphate dehydrogenases (mitochondrial and cytosolic) by NAD+/NADH redox method

Contact Info

Product

Resources

About