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An Extensive In vitro Approach in Development of Next Generation Drugs Against Trehalose-6-Phosphate Phosphatases Involved in Pathogenicity of Many Microorganisms
Abstract
Abstract
Trehalose-6-phosphate phosphatase (TPP) enzymes implicated in pathogenecity of number of bacteria, fungi and nematodes causing serious diseases such as tuberculosis, candedemias, aspergillosis, elephantiasis and parasitic ones. This study is an effort to target a common pathway critical to pathogenicity of different microbes but entirely absent in vertebrates and to come out with new generation drugs by an extensive in vitro approach. Five; Ribes nigrum, Aegle marmelos, Carica papaya, Ocimum basilicum and Costus pictus out of 21 hydroalcoholic (HA) extracts of medicinal plants showed significant anti-TPP activity against previously purified maize TPP by our group. To determine the active principles of above plants, extensive literature survey was done, and 17 phytochemicals were screened out based on their medicinal properties. TPP-phytochemical complexes of Rutin, Carpaine, Stigmasterol, β-Caryophyllene and α-Eudesmol (at least one best phytochemical from each of five plants showed inhibition of TPP) shows that leads were in tight binding with enzyme. In vitro studies of active principle reaffirm the inhibition of purified TPP by best of the leads. Further kinetic studies of Rutin on TPP indicated that it is a reversible competitive inhibitor with Kcat/Km values of 72.6 which was almost equal to similar Rice TPP. IC50 of Rutin was determined and found to be 0.6 mM and Ki value was 0.4 mM indicating the strength of enzyme-inhibitor complexes. Circular dichroism studies of TPP in absence and presence of Rutin hints at change in the secondary structure and overall 3D conformation of the protein indicated by increase in β-sheets and lowering of α-helix (Rutin increases the complexity of structure).
Keywords: trehalose-6-phosphate phosphatases (TPP), rutin, inhibitor kinetics, circular dichroism, in vitro
Cite this Article
Umesh HR, Ramesh KV. An Extensive In vitro Approach in Development of Next Generation Drugs Against Trehalose-6-Phosphate Phosphatases Involved in Pathogenicity of Many Microorganisms. Research and Reviews: Journal of Herbal Science. 2018; 7(1): 32–43p.
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DOI: https://doi.org/10.37591/rrjohs.v7i1.257
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