Optimization of riboflavin production by recombinant <Emphasis Type="Italic">Bacillus subtilis</Emphasis> RH44 using statistical designs |
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Authors: | Qiu-Li Wu Tao Chen Yu Gan Xun Chen Xue-Ming Zhao |
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Institution: | (1) Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People’s Republic of China |
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Abstract: | A sequential optimization strategy, based on statistical experimental designs, was used to enhance the production of riboflavin
by recombinant Bacillus subtilis RH44. In the first instance, the medium components were optimized in shake flask cultures. After preliminary experiments
of nitrogen source selection, the two-level Plackett–Burman (PB) design was implemented to screen medium components that significantly
influence riboflavin production. Among the 15 variables tested, glucose, NaNO3, K2HPO4, ZnSO4, and MnCl2 were identified as the most significant factors (confidence levels above 95%) for riboflavin production. The optimal values
of these five variables were determined by response surface methodology (RSM) based on the central composite design (CCD).
The validity of the model developed was verified, and the optimum medium led to a maximum riboflavin concentration of 6.65 g/l,
which was 44.3 and 76.4% higher than the improved medium and the basal medium, respectively. A glucose-limited fed-batch culture
profile in a 5-l fermentor was consequently designed according to the above optimum medium in shake flasks. A final riboflavin
concentration of 16.36 g/l was obtained in 48 h, which further verified the practicability of this optimum strategy. |
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Keywords: | Bacillus subtilis Riboflavin production Plackett– Burman design Central composite design Response surface methodology |
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