Screening, Identifying of Penicillium K-P Strain and Its Cellulase Producing Conditions

Cellulase production by Penicillium species are of greater interest in microbial enzyme technology. Penicillium sp. was cultivated in liquid culture medium at different carbon sources, nitrogen sources in different pH and temperature conditions. The strain of K-P with high cellulase activity was screened. The cellulase activity was 198 U/mL in the presence of fructose on day fifth. Maximum activity was recorded 154 U/mL in with the presence of ammonium nitrate on the fourth day. And maximum cellulase activity was obtained when the pH was 3.0 (129 U/mL) on day fourth. But the highest cellulase activity recorded (274 U/mL) in the presence of fructose, ammonium nitrate, pH 3.0 on the fifth day. The results showed the profiles of cellulase were produced maximum level according to which enzyme is most active in that particular environment

Detection and Analysis of Lysozyme Activity in some Tuberous Plants and Calotropis Procera’s Latex

Tuber extract of all the plant species used in the study exhibited lysozyme activity confirming the ubiquitous presence of lysozyme in plants. Among the different plants screened for protein content the tuber extract of Solanum tuberosum showed highest buffer soluble protein while tuber extract of Raphanus sativus showed the lowest protein content in sodium acetate buffer (50 Mm; pH 5.0). Tuber extract of Raphanus sativus showed highest lysozyme activity among all the plant species tested in this study and the activity was increased when the tuber was extracted with sodium phosphate buffer (50mM; pH 7.0). The lowest lysozyme was observed with tuber extract of Daucus carota in phosphate buffer (50mM; pH 7.0). The latex of the tropical species Calotropis procera is well known for being a rich source of the lysozyme. Lysozyme of Calotropis procera latex is not thermo labile. It did not lose much of its activity when the latex was incubated at different temperatures for 24 hours. A positive pointer for purification of this enzyme in future. Calotropis procera lysozyme can be was specifically isolated and purified from the whole latex with ammonium sulphate precipitation with 95% saturation. Calotropis procera lysozyme retained its activity even after precipitation with ammonium sulphate and dialysis and could hydrolyse the cell wall of Micrococcus lysodeikticus.Key words: Micrococcus lysodeikticus, Tuberous Plants, Latex, Ammonium sulphate precipitation, Polyacrylamide gel electrophoresis M. Sakthivel et al. Detection and Analysis of Lysozyme Activity in some Tuberous Plants and Calotropis Procera’s Latex. J Phytol 2/11 (2010) 65-72

Optimization of Culture Conditions for the Production of Extracellular Cellulase from Corynebacterium lipophiloflavum

Cellulolytic enzyme producing bacteria were isolated from decaying vegetables on nutrient agar plates. As many as fourteen different bacterial strains were isolated and they were screened for the production of cellulase enzyme on a medium containing carboxymethyl cellulose (0.5% w/v). A Congo red dye based qualitative assay was performed to identify cellulase producing bacteria isolated from the vegetables. Among the difference strains, after screening on CMC agar, one strain exhibited relatively higher cellulase activity and was identified as Cornybacterium lipophiloflavum by a bacteriologist. Culture conditions were optimized for the above bacterium in the production medium supplemented with different carbon and nitrogen sources, different pH and different concentrations of CMC. Glucose and yeast extract proved to be the suitable carbon and nitrogen sources while pH 7.0 was ideal for cellulase production by the bacterium. Carboxymethyl cellulose at 1% in the medium induced relatively higher cellulase activity. Electrophoretic analysis of the ammonium sulfate precipitated proteins showed three prominent bands and whose molecular masses were estimated as 60, 69 and 75 kDa.  Cellulase produced by Cornybacterium lipophiloflavum could efficiently remove dyes or inks from the pulp incubated either in the form of intact growing cell or with clarified supernatant. The enzyme also removed starch from the fabric incubated with culture supernatant suggesting the potential use of Cornybacterium lipophiloflavum cellulase in industrial applications

Optimization of Culture Conditions for the Production of Extracellular Cellulase from Enterococcus pseudoavium

Bacteria that inhabit decaying vegetables were isolated and screened for the production of the enzyme, amylase. One species identified as Enterococcus pseudoavium exhibited relatively higher amylase activity of the bacterial species tested. The bacterium produced the highest extracellular amylase at 72 h in a medium containing starch (1% w/v), galactose (0.5% w/v), and peptone (0.5% w/v), at pH 7.0. The extracellular enzyme was partially purified and its starch hydrolyzing potential was evaluated. The organism when grown with paper pulp deinked the pulp completely just after four days of growth. The bacterial cells immobilized in sodium alginate beads when cultured with paper pulp could decolorize it within 4 days. The extracellular amylase produced by Enterococcus pseudoavium effectively deinked and decolorized paper pulp within 4 days after incubation. The enzyme efficiently removed the starch present in fabric and thus it could be very well used as an ingredient in commercial detergent