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KMID : 0613820110210091205
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Journal of Life Science
2011 Volume.21 No. 9 p.1205 ~ p.1213
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Comparison of Statistical Methods for Optimization of Salts in Medium for Production of Carboxymethylcellulase of Bacillus amyloliquefaciens DL-3 by a Recombinant E. coli JM109/DL-3
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Lee You-Jung
Kim Hye-Jin
Gao Wa
Chung Chung-Han
Lee Jin-Woo
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Abstract
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The optimal concentrations of salts in medium for cell growth and the production of carboxymethylcellulase (CMCase) by a recombinant E. coli JM109/DL-3 were established using two statistical methods: orthogonal array method (OAM) and response surface method (RSM). The analysis of variance (ANOVA) of data based on OAM indicated that K₂HPO₄ gave maximum sum of square (S) and percentage contribution (P) for cell growth as well as production of CMCase. The optimal concentrations of K₂HPO₄, NaCl, MgSO₄·7H₂O, and (NH₄)₂SO₄ in medium for cell growth extracted by Qualitek-4 (W32b) Software were 10.0, 1.0, 0.2, and 0.6 g/l, respectively, whereas those for the production of CMCase by E. coli JM109/DL-3 were 5.0, 1.0, 0.4, and 0.6 g/l. The analysis of variance (ANOVA) resulting from RSM indicated that a highly significant salt for cell growth was K₂HPO₄ ("probe>F" less than 0.0001), whereas K₂HPO₄ and MgSO₄·7H₂O were significant for the production of CMCase. The optimal concentrations of K₂HPO₄, NaCl, MgSO₄·7H₂O, and (NH₄)₂SO₄ for cell growth extracted by Design Expert Software were 7.44, 1.08, 0.22, and 0.88 g/l, respectively, whereas those for production of CMCase were 5.84, 0.69, 0.28, and 0.54 g/l. The optimal concentrations of salts and their influences on cell growth and production of CMCase extracted by OAM were almost the same as those by RSM. Production of CMCase by a recombinant E. coli JM109/DL-3 under optimized concentration of salts was 1.93 times higher than that by Bacillus amyloliquifaciens DL-3.
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KEYWORD
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Carboxymethylcellulase, E. coli JM109, optimization, orthogonal array method, response surface method, salts
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