Exopolysaccharide production from Bacillus velezensis KY471306 using statistical experimental design

Abstract Exopolysaccharide (EPS) biopolymers produced by microorganisms play a crucial role in the environment such as health and bio-nanotechnology sectors, gelling agents in food and cosmetic industries in addition to bio-flocculants in the environmental sector as they are degradable, nontoxic. This study focuses on the improvement of EPS production through manipulation of different culture and environmental conditions using response surface methodology (RSM). Plackett-Burman design indicated that; molasses, yeast extract and incubation temperature are the most effective parameters. Box-Behnken RSM indicated that; the optimum concentration for each parameter was 12% (w/v) for molasses, 6 g/L yeast extract and 30 °C for incubation temperature. The most potent bacterial isolate was identified as Bacillus velezensis KY498625. After production, EPS was extracted, purified using DEAE-cellulose, identified using Fourier transform infrared (FTIR), gel permeation chromatography (GPC) and gas chromatography-mass spectroscopy (GC-MS). The result indicated that; it has molecular weight 1.14 × 105 D consisting of glucose, mannose and galactose.

Isolation and characterization of an antifungal peptide from fruiting bodies of edible mushroom Lentinus squarrosulus Mont

Aims: To isolate and characterize an antimicrobial peptide from fruiting bodies of Lentinus squarrosulus Mont., the Thai common edible mushroom. Methodology and results: Solid ammonium sulfate at 40-80% (w/v) final concentration was utilized to precipitate the proteins and further purified by ion exchange chromatography on DEAE-cellulose and gel filtration chromatography on Sephadex G-25. The peptide was adsorbed on DEAE-cellulose and Sephadex G-25. It appeared as a single band with a molecular mass of about 17 kDa (kilodalton) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Further investigation of antimicrobial properties of purified peptide revealed that it has no activity against both Gram positive and Gram negative bacteria. However, it exhibited strong antifungal activity against various species of fungal pathogen of human. Among the high sensitive strains, Trichophyton mentagrophytes, T. rubrum, and Candida tropicalis are clinical isolates. Moreover, the potency was found to be concentration dependent and comparable with Ketoconazole, the commercial antifungal drug. Conclusion, significance and impact study: In this work, the novel bioactive peptide from fruiting bodies of L. squarrosulus Mont. has been isolated. It shows potent activity against various clinical isolates of fungal pathogen of human. It may have potential for pharmaceutical application.

Isolation and characterization of extracellular thermostable alkaline phosphatase enzyme from bacillus spp.

Alkaline phosphatase (E C 3.1.3.1) belongs to the class of hydrolases and catalyzes the alkaline hydrolysis of a number of phosphoric acid esters, nucleotides etc. Alkaline phosphatase was produced from Bacillus spp, isolated from soil samples. The Bacillus spp. was identified by staining and standard biochemical tests after which screening was done using modified Pikovoskaya’s agar method. Production of alkaline phosphatase using different substrates like calcium phosphate along with casein, starch, glucose and glutamic acid was carried out. High activity was found in calcium phosphate along with the casein. The specific activity of the crude extract was found to be 0.825U and it was subjected to purification by DEAE-Cellulose ion exchange chromatography. Finally,36% recovery was obtained. The molar mass was estimated by using 10% SDS-PAGE and was found to be approximately 84 KD. The optimum activity was at pH 8.8 and temperature of 650C. Alkaline phosphatase activity was enhanced by Mg2++ upto 66% and 80% activity was inhibited by EDTA. Alkaline Phosphatase activity was also confirmed by zymography using malachite green staining method.

Purification and Characterization of Intracellular Cellulase from Aspergillus oryzae ITCC-4857.01

Purification and characterization of intracellular cellulase produced by A. oryzae ITCC-4857.01 are reported. The enzyme was purified by ion-exchange chromatography using DEAE-cellulose followed by Gel filtration. The purification achieved was 41 fold from the crude extract with yield of 27%. The purified enzyme showed single band on poly acrylamide gel. The molecular weight as determined by SDS-PAGE and gel filtration was 38 KDa and 38.6 KDa respectively and contained only one subunit. The enzyme is glycoprotien as nature and contained 0.67% neutral sugar. The apparent Km value of the enzyme against cellulose was 0.83%. The enzyme showed the highest relative ativities on CMC followed by avicel, salicin and filter paper. The optimum pH of activity was 5.5 and very slight activity was observed at or above pH 7.5 as well as bellow pH 3.5. The optimum tempreture of the activity was 45degrees C and the highest activity was exhibited in 35 to 45degrees C. The enzyme lost their activities almost completely (95~100%) at 80 degrees C or above and as well as bellow 25degrees C.

Purificación de la proteína banda 3 de eritrocitos humanos (AE 1) / Purification of band 3 protein from human erythrocytes (AE 1)

Se purifica la proteína banda 3 a partir de membranas de eritrocitos humanos con rapidez y eficiencia, mediante una combinación de cromatografía de intercambio iónico en DEAE-celulosa y de afinidad mediante el empleo de una columna pCMB-Sepharose. La obtención de banda 3 permitirá investigar su posible participación en los fenómenos oclusivos en la drepanocitosis.

The band 3 protein is purified starting form the membranes of human erythrocytes with celerity and efficiency by a combination of DEAE-cellulose ion-exchange and affinity chromatography by using a pCMB-Sepharose column. The obtention of band 3 will allow to investigate its possible participation in the occlusive phenomena in drepanocytosis.