Asma Ansari

Degradation of complex carbohydrate: immobilization of pectinase from Bacillus licheniformis KIBGE-IB21 using calcium alginate as a support.

Pectinases are heterogeneous group of enzymes that catalyse the hydrolysis of pectin substances which is responsible for the turbidity and undesirable cloudiness in fruits juices. In current study, partially purified pectinase from Bacillus licheniformis KIBGE-IB21 was immobilized in calcium alginate beads. The effect of sodium alginate and calcium chloride concentration on immobilization was studied and it was found that the optimal sodium alginate and calcium chloride concentration was 3.0% and 0.2 M, respectively. It was found that immobilization increases the optimal reaction time for pectin degradation from 5 to 10 min and temperature from 45 to 55°C, whereas, the optimal pH remained same with reference to free enzyme. Thermal stability of enzyme increased after immobilization and immobilized pectinase retained more than 80% of its initial activity after 5 days at 30°C as compared with free enzyme which showed only 30% of residual activity. The immobilized enzyme also exhibited good operational stability and 65% of its initial activity was observed during third cycle. In term of pectinase immobilization efficiency and stability, this calcium alginate beads approach seemed to permit good results and can be used to make a bioreactor for various applications in food industries.

Production of xylan degrading endo-1, 4-β-xylanase from thermophilic Geobacillus stearothermophilus KIBGE-IB29

Abstract Xylan degrading bacterial strain was isolated from soil and identified as Geobacillus stearothermophilus KIBGE-IB29 on the basis of morphological, biochemical and 16S rDNA sequence analysis. Optimization of medium and culture conditions in submerged fermentation was investigated for maximum endo-1, 4-β-xylanase production. High yield of xylan degrading endo-1, 4-β-xylanase was achieved at 60°C and pH-6.0 with 24h of fermentation. Maximum enzyme was produced using 0.5% xylan as a carbon source, 0.5% peptone, 0.2% yeast extract and 0.1% meat extract as nitrogen sources. Di-potassium hydrogen phosphate (0.25%), calcium chloride (0.01%), potassium hydrogen phosphate (0.05%) and ammonium sulfate (0.05%) were also incorporated in the fermentation medium to enhance the enzyme production.

Dextranase: Hyper production of dextran degrading enzyme from newly isolated strain of Bacillus licheniformis

Dextranase, 6-alpha-D-glucan 6-glucanohydrolase catalyzes the degradation of dextran (polymer of D-glucose) in to low molecular weight fractions. Dextranolytic bacterial strains were isolated from various natural sources and plate assay methods were developed for screening of highest extracellular dextranase producing isolate. Bacillus licheniformis, identified on the basis of taxonomic characterization was subjected to UV radiation and highest enzyme producing mutant obtained led to 7 times more dextranase production than wild. Optimization of major physico-chemical parameters affecting enzyme production; including medium composition, pH, cultivation time and temperature revealed that maximum enzyme production was obtained in a self designed medium (pH 6.0) containing 1% Dextran 5000 Da, after 24 h culture incubation at 37 °C. Dextranase reported in this study is of great commercial importance as it is strictly inducible in nature and B. licheniformis being non-pathogenic removes the safety concerns associated with production of dextran fractions for clinical and pharmaceutical usage.

Purification, characterization and end product analysis of dextran degrading endodextranase from Bacillus licheniformis KIBGE-IB25

Degradation of high molecular weight dextran for obtaining low molecular weight dextran is based on the hydrolysis using chemical and enzymatic methods. Current research study focused on production, purification and characterization of dextranase from a newly isolated strain of Bacillus licheniformis KIBGE-IB25. Dextranase was purified up to 36 folds with specific activity of 1405 U/mg and molecular weight of 158 kDa. It was found that enzyme performs optimum cleavage of dextran (5000 Da, 0.5%) at 35 °C in 15 min at pH 4.5 with a Km and Vmax of 0.374 mg/ml and 182 μmol/min, respectively. Relative amino acid composition analysis of purified enzyme suggested the presence of higher number of hydrophobic, acidic and glycosylation promoting amino acids. The N-terminal sequence of dextranase KIBGE-IB25 was AYTVTLYLQG. It exhibited distinct amino acid sequence yet shared some inherent characteristics with glycosyl hydrolases (GH) family 49 and also testified the presence of O-glycosylation at N-terminal end.

Characterization and interplay of bacteriocin and exopolysaccharide-mediated silver nanoparticles as an antibacterial agent

Metallic nanoparticles have a substantial scientific interest because of their distinctive physicochemical and antimicrobial properties and the emergence of multidrug resistant pathogens could unlock the potential of nanoparticles to combat infectious diseases. The aim of the current study is to enhance the antibacterial potential of purified bacteriocin by combining bacteriocin and antibacterial silver nanoparticles (AgNPs). Hence, the interaction of natural antimicrobial compounds and antibacterial nanoparticles can be used as a potential tool for combating infectious diseases. In this study, a green, simple and effective approach is used to synthesize antibacterial AgNPs using fungal exopolysaccharide as both a reducing and stabilizing agent. The AgNPs were characterized by spectroscopic analysis, Scanning Electron Microscopy (SEM), Energy Dispersive X-ray spectroscopy (EDX) and Dynamic Light Scattering (DLS). Furthermore, the synergistic effect of bacteriocin-AgNPs was determined against pathogenic strains. The histogram of AgNPs indicated well-dispersed, stabilized and negatively charged particles with variable size distribution. The combination of bacteriocin with nanoparticles found to be more effective due to broad antibacterial potential with possibly lower doses. The current study is imperative to provide an alternative for the chemical synthesis of silver nanoparticles. It showed environmental friendly and cost effective green synthesis of antibacterial nanoparticles.

Bacteriocin from LAB for Medical and Health Applications

The emergence of serious issues of multidrug resistance in the past few years forced the consideration of bacteriocins for combating infections. Numerous concerns have been raised against increased bacterial resistance toward effective drugs and become a debated issue all over the world. Alongside, there is an increase in consumer demand for the antimicrobial compounds isolated or derived from natural sources. Production of antimicrobial compounds is considered as a ubiquitous anti-competitor strategy in microbial ecosystem. Research on antimicrobial compounds with a special interest on bacteriocins is opening a door of a new age. Bacteriocins play an immense role in different industries to overcome various unrestrained environmental issues. Many researchers are now focusing on the bacteriocins of lactic acid bacteria (LAB) with plenty of applications not only in food industries but also in medical and health applications. Their infrequent and targeted use leads to the reduction in the emergence of drug resistance by microbes. Currently, bacteriocins produced by LAB are extensively studied due to their generally recognized as safe (GRAS) status. Various species of LAB are reported to have therapeutic properties that confer beneficial effects on human and animal health. The public health dilemma of drug resistance can be resolved by the discovery of new antimicrobial compounds having target-specific inhibition especially against multidrug-resistant organisms. Consequently, the pool of effective drugs could be available all the time to control newly emerging drug resistance in bacteria.