Gene analysis,optimized production and property of marine lipase from <Emphasis Type="Italic">Bacillus pumilus</Emphasis> B106 associated with South China Sea sponge <Emphasis Type="Italic">Halichondria rugosa</Emphasis> |
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Authors: | Hongzhen Zhang Fengli Zhang Zhiyong Li |
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Institution: | (1) Laboratory of Marine Biotechnology, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 800 Dong-Chuan Road, 200240 Shanghai, People’s Republic of China;(2) Key Laboratory of Microbial Metabolism, Ministry of Education, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 800 Dong-Chuan Road, 200240 Shanghai, People’s Republic of China |
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Abstract: | Gene cloning, optimized production and property of marine lipase from Bacillus pumilus B106 associated with South China Sea sponge Halichondria rugosa were investigated in this paper. A lipase gene with whole ORF encoding 215 amino acids was obtained by PCR, protein domain
prediction suggested that the deduced lipase belongs to α/β hydrolases family. Based on single factor Seriatim-Factorial test
and Plackett–Burman experimental design, the optimal medium consisted of (per l) 12.5 ml maize oil, 5.0 g beef extract, 2.0 g
PO4
3− (0.6 g KH2PO4, 1.4 g K2HPO4), 17.15 g Mg2+, 5.0 g yeast extract, 2.282 g CaCl2 and 5.0 ml Tween80 with artificial sea water. Using this optimum medium, lipase activity and cell concentration were increased
by 3.54- and 1.31-fold over that of the basal medium, respectively. This lipase showed tolerance to high salinity, pH and
temperature. About 10–20% methanol exhibited a stimulatory effect on the lipase activity, while activity was inhibited by
30–40% methanol, 2-propanol, DMSO, and ethanol. This study provides a valuable resource for marine lipase production and extends
our understanding of the possible role of sponge-associated bacteria in the biotransformation of chemical compounds for the
sponge host. |
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Keywords: | Halichondria rugosa Bacillus pumilus Lipase Gene cloning Statistical optimization Property |
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