Eudesmol-A promising inhibitor for glucosyltransferase: Docking and Molecular dynamics study

The loss of natural teeth can be avoided by invoking the molecular signal behind teeth regeneration. The destruction of the connective tissues is mainly due to bacterial origin which reacts to dental caries, a multifactorial disease. Glycosyl transferase is the enzyme which is involved in the glycosidic linkage. Glucosyltransferase inactivation reduces dental caries. This enzyme is a crucial virulence factor of Streptococcus mutans, a major pathogen that causes dental caries. In this present work, screening was done with library of anti-oxidant and anti-inflammatory molecules against the crystal structure of the target protein. Based on the predicted binding affinities, small molecules were selected and evaluated for their activity. Further, attempts were done to evaluate the toxicity of the lead compounds and compounds with no toxicity and good binding affinity were subjected for simulation and compared with reference complex. The potential energy of Glycosyl transferase-Eudesmol (proposed compound) (-1500 kj/mol) indicates its higher stability as compared to Glycosyl tranferase-G43 (reference) complex (-1100kj/mol). The inactives and actives compound for Glycosyl transferase was predicted from DeepScreening server.

Identification of Two Novel Frameshift Mutations in Exostosin 1 in Two Families with Multiple Osteochondromas

Multiple osteochondromas (MO) is an autosomal dominant hereditary disorder, which typically manifests as skeletal dysplasia, mainly involving long bones and knees, ankles, elbows, wrists, shoulders, and pelvis. Previous studies have demonstrated that mutations in exostosin glycosyl transferase-1 (<i>EXT1</i>) and exostosin glycosyl transferase-2 (<i>EXT2</i>) were the main cause of MO. In this study, we enrolled 2 families with MO. Sanger sequencing revealed 2 novel frameshift mutations – c.1432_1433insCCCCCCT; p.Lys479Profs*44 and c.1431_1431delC; p.S478PfsX10 – in the <i>EXT1</i> gene detected in 2 families, respectively. Both novel mutations, located in the conserved domain of EXT1 and predicted to be disease causing by informatics programs, were absent in our 200 control cohorts and other public databases. Our study expanded the spectrum of <i>EXT1</i> mutations and contributed to genetic diagnosis and counseling of patients with MO.

Deletion of a Peptidylprolyl Isomerase Gene Results in the Inability of Caldicellulosiruptor bescii To Grow on Crystalline Cellulose without Affecting Protein Glycosylation or Growth on Soluble Substrates

ABSTRACT Caldicellulosiruptor bescii secretes a large number of complementary multifunctional enzymes with unique activities for biomass deconstruction. The most abundant enzymes in the C. bescii secretome are found in a unique gene cluster containing a glycosyl transferase (GT39) and a putative peptidyl prolyl cis-trans isomerase. Deletion of the glycosyl transferase in this cluster resulted in loss of detectable protein glycosylation in C. bescii, and its activity has been shown to be responsible for the glycosylation of the proline-threonine rich linkers found in many of the multifunctional cellulases. The presence of a putative peptidyl prolyl cis-trans isomerase within this gene cluster suggested that it might also play a role in cellulase modification. Here, we identify this gene as a putative prsA prolyl cis-trans isomerase. Deletion of prsA2 leads to the inability of C. bescii to grow on insoluble substrates such as Avicel, the model cellulose substrate, while exhibiting no differences in phenotype with the wild-type strain on soluble substrates. Finally, we provide evidence that the prsA2 gene is likely needed to increase solubility of multifunctional cellulases and that this unique gene cluster was likely acquired by members of the Caldicellulosiruptor genus with a group of genes to optimize the production and activity of multifunctional cellulases. IMPORTANCE Caldicellulosiruptor has the ability to digest complex plant biomass without pretreatment and have been engineered to convert biomass, a sustainable, carbon neutral substrate, to fuels. Their strategy for deconstructing plant cell walls relies on an interesting class of cellulases consisting of multiple catalytic modules connected by linker regions and carbohydrate binding modules. The best studied of these enzymes, CelA, has a unique deconstruction mechanism. CelA is located in a cluster of genes that likely allows for optimal expression, secretion, and activity. One of the genes in this cluster is a putative isomerase that modifies the CelA protein. In higher eukaryotes, these isomerases are essential for the proper folding of glycoproteins in the endoplasmic reticulum, but little is known about the role of isomerization in cellulase activity. We show that the stability and activity of CelA is dependent on the activity of this isomerase.