Journal of Probiotics & Health
Open Access

Protein profiling, PGH activity and enzymatic studies in extracellular fluid of a potential probiotic microbe: An attempt towards development of a probiotic

4^th International Conference and Exhibition on Probiotics, Functional and Baby Foods

November 03-05, 2015 Valencia, Spain

Jasbir Singh, Suman Dhanda, Dimpi Gandhi, Preeti Chanalia, Davender Kumar, Dimple Nanda and Tannu Talwar

Kurukshetra University, India

Posters-Accepted Abstracts: J Prob Health

Abstract :

Though there are insufficient reports on how probiotics benefit yet providing enzymes to host may be one of the mechanism of probiotic action. Pediococcus acidilactici is harmless lactic acid bacteria with probiotic potential, so extracellular proteases of Pediococcus acidilactici were analysed. Protein profiles studied by gradient SDS-PAGE revealed that extracellular polypeptides were in the range of 35-75 kDa whereas membrane polypeptides were distributed in the range of 35-43 kDa. Minimum inhibitory concentration (MIC) of vancomycin for P. acidilactici was found to be 3.0 mg/ml. Vancomycin resistant P. acidilactici expressed higher level of polypeptides of 70-100 kDa which may be target of interaction studies. Since it is a facultative anaerobe, peptidoglycan hydrolase (PGH) studies in extracellular and intracellular fractions under anaerobic conditions were confirmed by zymographic and turbidometric method against Staphylococcus albus, Lactobacillus collinoides, Lactobacillus paracasei and Bacillus cereus. Intracellular PGH was comparable to 50 �¼g/ml of lysozyme at same substrate concentration. PGH activity suggests an alternative therapy to antibiotics and studies need to be extended to aerobic conditions. Extracellular Lys-Ala-4�²NA hydrolysing enzyme was purified by ammonium sulphate fractionation (0-80%) and gel filtration chromatography. The enzyme optimally worked at pH 7.0 at 37�° C. This enzyme preferably hydrolysed Lys-Ala-4�²NA with Km of 60 �¼M. Enzyme inhibition by PMSF and DEPC suggest it to be a serine protease with involvement of histidine residues in enzyme catalysis. This enzyme is reported to regulate the cell cycle but its function is yet to be explored in microbes. All these studies are important from pharmaceutical point of view towards the development of P. acidilactici as probiotic.

Biography :

Email: jasbirdhanda@gmail.com