Optimization of culture medium components for agarase-producing marine bacterial strain Brevundimonas sp. DT-7
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    Abstract:

    The activity of agarase depended on the bacterial growth conditions such as growth medium formula. In this study, we used marine bacterial strain Brevundimonas sp. DT-7 as an extracellular agarase producer and optimized the growth medium formula to produce maximum agarase that can be used for producing oligosaccharides from agar on a commercial scale in the future. Eight components in the bacterial culture medium (X1 agar powder,X2 NaCl, X3 FePO4X4 CaCl2X5 K2HPO4X6 FeSO4·7H2O,X7 yeast extract, X8 peptone) were selected for the optimization. PlackettBurman design was used in the experiments and the significance of each component (X1-X8) contributing to activity of agarase (Y1) was determined. The significant levels were defined at P<0.05. The data show that among the eight components, three medium components (X1 agar powder, X4 CaCl2 and X8 peptone) are significant factors for production of agarase from DT7. The three significant factors (X1 agar powder, X4 CaCl2 and X8 peptone) were selected for further analysis using BoxBehnken design. A regression model was obtained by response surface regression method (SAS 8.2) as follows: Y1=499.6+19.45X1+13.99X4-1.213X8-33.16X12-5.3X1X4-3.45X1X8-39.64X42-10.23X4X8-15.79X82. The results demonstrate the optimum culture medium components (w/v): 0.536 % agar powder, 2.5% NaCl, 0.1% FePO4, 0.052% CaCl2, 0.1% K2HPO4,0.03% FeSO4·7H2O, 1% yeast extract, and 0.484% peptone, respectively. Using those optimized culture components and combined with other culture conditions [50 mL culture medium in 250 mL flask, initial pH 7.5, shaken at 120 r/min, incubated at (23±1)℃ for 28 h], the agarase activity of 503.3 U/mL was obtained, which was significantly higher than that of 372.0 U/mL when the culture medium was not optimized. We conclude that (1) using SAS software is a very effective and quick way to optimize a bacterial growth culture medium; (2) the culture medium optimized in this study is simply formulated and can be used for production of high activity of agarase at low cost; (3) with further modification and improvement, Brevundimonas sp. DT7 can be a source bacterium for production of agarase on a commercial scale. This is a preliminary study. Further research will be required to evaluate Brevundimonas sp. DT-7 in commercial application.

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桑卫国,ZHANG Zong-ming. Optimization of culture medium components for agarase-producing marine bacterial strain Brevundimonas sp. DT-7[J]. Journal of Fisheries of China,2009,33(3):511~518

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History
  • Received:October 23,2008
  • Revised:February 22,2009
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  • Online: May 07,2009
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