Characterization and optimization of extracellular alkaline lipase production by Alcaligenes sp. Using stearic acid as carbon source |
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Authors: | Masahiro Mori Ehsan Ali Dongning Du Enoch Y Park |
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Institution: | (1) Laboratory of Biotechnology, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan;(2) Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan |
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Abstract: | The aim of this study was to improve the production of an extracellular alkaline lipase from Alcaligenes sp. (ATCC 31371) by optimization of the culture medium, for economic production of biodiesel from waste vegetable oil. A
number of carbon sources including different types of starch, sugar, sugar alcohol, organic acids, and surfactants were investigated.
Polyoxyethylene (20) sorbitan tristearate, whose side chain is stearic acid, was the most effective carbon source for lipase
production. Box-Behnken experimental design was used for three factors (soy protein, sodium nitrate, and stearic acid) and
the optimal composition for maximum lipase production (1.7-fold enhancement) was established as soy protein 4.07%, sodium
nitrate 0.17%, and stearic acid 0.28% at 28°C with an agitation rate of 220 rpm for 24 h. The enzyme was purified to homogeneity
and the recovery of the lipase activity was 7.8% with a 30-fold purification. The estimated molecular size of the protein
determined by SDS-PAGE was 33 kDa. The optimum pH and temperature of the purified lipase was 8.5 and 40°C, respectively. The
purified enzyme was stable in the pH range of 6.0 and 9.5 and in the temperature range of 20 and 50°C. |
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Keywords: | Alcaligenes sp alkaline lipase medium optimization waste activated bleaching earth bio-refinery |
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