SCREENING AND EXTRACTION OF POTENTIAL INHIBITORS FOR GLUCANSUCRASE FROM HERBAL SOURCES TO TACKLE DENTAL CARIES

PROJECT REFERENCE NO.: 39S_R_MTECH_042

COLLEGE : SIR M VISVESVARAYA IINSTITUTE OF TECHNOLOGY, BENGALURU BRANCH : DEPARTMENT OF BIOTECHNOLOGY GUIDES : MS. PRIYA NARAYAN STUDENTS : MS. SRINIDHI UPENDRA BHUJANG

KEYWORDS: Dental caries, Glucansucrase (3AIC), Salivary amylase (1SMD), Streptococcus mutans.

INTRODUCTION: Dental caries is most common childhood disease worldwide. It is still a major problem in most industrialized countries, affecting 60-90% of school children and a majority of adults. Access to oral health services in developing countries is limited, and teeth are often left untreated or are extracted due to pain and fourth most expensive disease to treat in most industrialized countries. By age 15, approximately 60% of globe will have experienced tooth decay.

The main criteria required for caries formation: A tooth surface (enamel or dentin), caries-causing bacteria, fermentable carbohydrates (such as sucrose). These criteria are not always enough to cause the disease and a sheltered environment promoting development of a cariogenic biofilm is required.The bacteria most responsible for dental cavities are the mutans streptococci, most prominently Streptococcus spp(mutans, sobrinus) and lactobacillus spp.

OBJECTIVES: Initially bioinformatics screening and confirmation is done which helps in screening the compounds which binds to glucansucrase and not to salivary amylase, i.e., Screening/insilico analysis of inhibitor molecules with Glucansucrase. 1. Extraction of compound from the particular herbal source. 2. Inhibition plate assay of the extract on biofilm and glucansucrase action. METHODOLOGY: By using Pymol, multalin, consurf we were able to analyse the binding site and its similarities between glucansucrase and salivary amylase and also studied the disorderness of the enzyme glucansucrase by using Psipred to know about the secondary structure. Nextly we went with docking of about 4500 molecule available in ZINC docking library to Glucansucrase using Pyrx and autodock vina. Right now we are doing assay by crude extract of the herbal source where our compounds are present, by extracting it by solvent extraction and then evaporating and using it for initial assay.

RESULTS AND CONCLUSIONS: Selected top 10 molecules which docked with Glucansucrase with better binding energy(-10Kcal/mol) and least with salivary amylase. The energy of binding of our drug molecules, when compared sugar, is showing better binding capacity, implying that it blocks the binding of sugar molecule to Glucansucrase, and thus preventing formation of biofilms. Based on this study we have also got that these screened compounds are inhibiting the enzymes when it was seen in biochemical analysis showing decrease in the amount of production of glucose. As glucose is produce if sucrose is broken down only when glucansucrase acts on it.

SCOPE FOR FUTURE WORK: 1. Extraction of pure compound that is docked from the respective sources. 2. As saying the compound on biofilm inhibition and enzymes inhibition and not on the organism. 3. Assaying of capability of the compound for introduction into food and toothpastes.