A soil bacterium, Bacillus subtilis, isolated from the rhizosphere of Chilli, showed high antagonistic activity against Colletotrichum gloeosporioides OGC1. A clear inhibition zone of 0.5–1 cm was observed in dual plate assay. Microscopic observations showed a clear hyphal lysis and degradation of fungal cell wall. In dual liquid cultures, the B. subtilis strain inhibited the C. gloeosporioides up to 100 % in terms of dry weight. This strain also produced a clear halo region on chitin agar medium plates containing 0.5 % colloidal chitin, indicating that it excretes chitinase. The strain also produced other mycolytic enzymes—glucanase and cellulase, demonstrated by a clear zone of hydrolysis of yeast cell wall glucan (YCW 0.1 % v/v) and carboxymethylcellulose (CMC 0.1 % v/v). In liquid cultures, the strain showed appreciable levels of chitinase, glucanase and cellulase activities and hydrolytic activity with C. gloeosporioides OGC1 mycelia as the substrate. The role of the B. subtilis strain in suppressing the fungal growth in vitro was studied in comparison with a UV mutant of that strain, which lacked both antagonistic and hydrolytic activity. The mycolytic enzyme mediated antagonism of B. subtilis was further demonstrated by heat inactivation (70–100 °C), treatment with trypsin and TCA of the crude enzyme extract which lacked antifungal property also. Treatment of the chilli seeds with Bacillus sp. culture showed 100 % germination index similar to the untreated seeds. The treatment of the seed with co-inoculation of the pathogen with Bacillus sp. culture showed 65 % reduction in disease incidence by the treatment as compared to the seed treated with pathogen alone (77.5 %).
The study identified the innate enzymatic potential (amylase) of the PHB producing strain B.thuringiensis IAM 12077 and explored the same for cost-effective production of PHB using agrowastes, eliminating the need for pretreatment (acid hydrolysis and/or commercial enzyme
An extracellular lipase with 23,666.66 U/ml/min activity was produced by Aspergillus tamarii JGIF06 under submerged fermentation in mineral salt medium containing coconut oil (2.5 % v/v), tryptone (2 % w/v) and ammonium chloride (2 % w/v), with initial pH of 5 ± 0.2, incubated at 25 °C for 7 days on a rotary shaker at 120 rpm. A 7.9-fold increase in lipase-specific activity was recorded after purification by DEAE Sepharose ion exchange and Sephadex G200 column chromatography. The apparent molecular mass of this enzyme was revealed as 50 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The optimal lipase activity was recorded at pH 4 and 37 °C. The enzyme revealed broad specificity towards different vegetable oils. The K
m and V
max of the lipase on olive oil was found to be 330.4 mg and 53,690 U/ml/min, respectively. The lipase activity was stable in the presence of surfactants such as cetrimonium bromide, sodium dodecyl sulphate and Tween 80, and metal ions and reagents such as Ca2+, Ba2+ and 2-mercaptoethanol. However, the activity was greatly reduced in the presence of organic solvents such as chloroform. The stain removal potential of the crude lipase was determined on polycotton fabric pieces stained with peanut oil. Lipase added to cold water alone significantly enhanced the removal of stain by 152 %. The addition of lipase also improved the stain removal efficiency of a commercially available detergent in the presence of either cold (25 ± 2 °C) or hot (65 ± 2 °C) water. The current findings suggest the potentiality of this enzyme for energy-efficient biocatalytic application.
The present study was undertaken to study the statistical optimization of medium components for improved Indole-3-acetic acid (IAA) production by Pseudomonas aeruginosa. Colorimetric analysis showed maximum IAA (132 µg mL-1) in the medium supplemented with tryptophan (0.5 g L-1). The maximum production was achieved after 96 h of incubation. Yeast extract, tryptophan and EDTA were identified as significant components influencing IAA production by Pseudomonas aeruginosa, using the Plackett-Burman method. The statistical optimization approach led to the production of 318 µg mL-1 of IAA within 24 h of incubation. Statistical approach was found to be very effective in optimizing the medium components in a manageable number of experimental runs with overall 2.4 fold increase in IAA production. TLC and GC-MS analysis further confirmed the IAA production in the cell filtrates of the strain. GC-MS analysis and tryptophan side chain oxidase confirmed the existence of atleast 2 possible pathways for IAA by this strain. Inoculation of P. aeruginosa culture filtrate enhanced seed germination (82.4%) and increase in root length and shoot length (~ 2.6 and ~ 1.1 folds over the control) of cowpea seeds over the control treatment under pot culture conditions.
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