Jeeva, P. | Search bioRxiv

bioRxiv2023

Unraveling the Potential of Epicatechin Gallate in Targeting Aberrant Cardiac Ca2+ Signaling Protein

The cardiovascular sarcoplasmic reticulum (SR) calcium (Ca2+) ATPase is an imperative determinant of cardiac functionality. In addition, anomalies in Ca2+ handling protein and atypical energy metabolism are inherent in heart failure (HF). Moreover, Ca2+ overload in SR leads to mitochondrial matrix Ca2+ overload, which can trigger the generation of Reactive Oxygen Species (ROS), culminating in the triggering of the Permeability Transition Pore (PTP) and Cytochrome C release, resulting in apoptosis that leads to arrhythmias and numerous disorders. Although proteins involved in the molecular mechanism of Ca2+ dysfunction regarding mitochondrial dysfunction remains elusive, this study aims to assess the major Ca2+ handling proteins which may be involved in the Ca2+ malfunction that causes mitochondrial dysfunction and predicting the most effective drug by targeting the analyzed Ca2+ handling proteins through various insilico analyses.Thirteen proteins absorbed from interaction analysis were docked with four optimal phytochemicals from Crataegus oxyacantha (COC) to identify the potential agonist/antagonist against those anomalies causing Ca2+ handling signaling proteins. Furthermore,The ADMET profile of tyramine, vitexin, epicatechin, and epicatechin gallate was acclimated to evaluate potential drugability utilizing QikProp by Schrodinger.Keeping this in view, critical molecular docking evaluations were performed using Glide (Maestro), autodock, and autodock vina.Based on the results of 156 dockings by Maestro, auto-dock, and auto-dock vina, PKA C with epicatechin gallate exhibits good interaction. Therefore, a 2000ns molecular dynamics (MD) simulation was utilized to assess the feasible phytochemical epicatechin gallate - PKA C complex binding stability utilizing Desmond. As a result, the molecular dynamics simulation study confirmed that epicatechin gallate from COC has high possibilities to inhibit the aberrant cardiac Ca2+ signaling proteins due to its conformational rigidity.

bioinformatics↗

bioRxiv2022

Metabolic Engineering Of Lactococcus Lactis For The Production Of Heparosan

Heparosan is a precursor molecule for the widely used anticoagulant heparin, which also has other uses such as certain drug delivery applications and as a scaffold for tissue engineering in biomaterials. Traditionally, pathogenic bacteria such as E.Coli have been used as a host to produce heparosan as an alternative to animal and chemoenzymatic synthesis. Using GRAS status organisms like Lactococcus Lactis as the host for production of heparosan provides a safe alternative as well as being a well-established organism for genetic manipulation and reengineering. In this study, a functional heparosan synthesis pathway was successfully expressed in Lactococcus Lactis by the expression of E.coli K5 genes KfiA and KfiC, along with the overexpression of ugd, glmu and pgma genes present natively in the host organism. The genes were activated using the tightly controlled NICE expression system. The genes were cloned into plasmid p8148 and transformed into two strains, Lactococcus Lactis NZ9000 and Lactococcus Lactis NZ9020, totaling six different recombinant strains were created using these two hosts and various combinations of the heterologous genes. The recombinant Lactococcus Lactis SH6 strain, expressing the genes ugd-KfiA-KfiC-pgma yielded a maximum concentration of 754 mg/l in batch bioreactor experiments and the titer was increased to 1263 mg/l in fed-batch fermentation. NMR imaging successfully determined that the structure of the product derived from Lactococcus Lactis was indeed similar to E.coli heparosan. The molecular weight of heparosan varied from 10-20 KDa, indicating its potential use for chemoenzymatic heparin biosynthesis.

bioengineering↗