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The culture conditions for the production of carrageenase were optimized using one-factor-at-a-time method combined with orthogonal array design. With the one-factor-at-a-time method revealed optimal conditions for carrageenase production were 24 h of fermentation period, 28 °C incubation temperature at pH 8.0 with NaNO3 as nitrogen source and carrageenan as carbon source in MMS media. Further optimization of carrgeenase production by using orthogonal experimental design L9 (34) with four factors, temperature, pH, NH4NO3 and carrageenan with their relevant levels revealed optimised conditions for carrageenase production were temperature of 28 °C, pH 8.0, 2 g L-1 NaNO3 and 2 g L-1 carrageenan. The order of the factors affecting the fermentation process was found to be temperature > pH > NaNO3 > carrageenan. The temperature played a significant role on the carrageenase production. Higher carrageenase yield with activity of 0.542 ±0.045 U ml-1 was obtained in the optimised medium when compared to those of basal medium. Carrageenase hydrolysed products of carrageenan were identified by LC-ESI-MS as neocarrabiose, neocarrabiose-4 sulfate, neocarratetraose, neocarratetraose-4 sulfate, anhydrogalactose, galactose, galactose-4 sulphate and sulphate.
An agar degrading bacterium Pseudomonas aeruginosa AG LSL-11 was acclimatized to alginate for the production of alginase. Production parameters such as pH, temperature, influence of simple carbohydrates and nitrogen sources, and effect of NaCl on growth and alginase production were carried out. Maximum growth was observed at pH 9.0 and 35 °C, while alginase was produced optimally at pH 9.0 and 30 °C. The alginase produced by Pseudomonas aeruginosa AG LSL-11 was inducible by alginate, and repressed by other simple sugar when supplemented along with alginate in the medium. The bacterium did not require NaCl for growth and production of alginase. The activity staining of partially purified culture supernatant after native PAGE revealed the presence of a single alginase.
The culture conditions for the production of extracellular agarase by Pseudomonas aeruginosa ZSL-2 were optimized using One-Factor-At-A-Time combined with orthogonal array design. One-Factor-At-A-Time method investigates the effect of time, temperature, NaCl, carbon sources, nitrogen sources and pH on agarase production. The optimized culture conditions obtained from the statistical analysis were temperature of 30 °C, pH 8.5, NH4NO3 2 g L-1 and agar 3 g L-1. The L9 orthogonal array design was used to select the fermentation parameters influencing the yield of agarase. The order of the factors affecting the fermentation process was found to be NH4NO3 > pH > agar > temperature, with temperature playing a significant role on the agarase production (p < 0.10). The higher yields than those in basal media culture were obtained in the final optimized medium with activity of 0.439 ± 0.013 U ml-1. Extracellular agarase hydrolysed agar into a range of oligosaccharides which were analysed by LC-ESI-MS spectrometry as anhydrogalactose, galactose, agarobiose, agarotetrose and agarohexaose.
Effect of plant growth regulators on seed germination and seedling characters in terms of root length and shoot length. The seeds of sesame variety TVM – 1 were treated with different concentration of gibberlic acid (1.0, 2.0, 3.0 and 4.0 mg/l) and indole acetic acid (1.0, 2.0, 3.0 and 4.0 mg/l). From the results, it was observed that the GA3 and IAA at 2.0mg/l had regulatory effect on seed germination and seedling characters. Maximum germination, root length and shoot length was observed at 2.0mg/l in GA3 and 2.0mg/l in IAA than control. And GA3 at 2.0gm/l was found to be more efficient to modify seed germination and seedling characters when compared to IAA and control.
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