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KMID : 1094720110160061144
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Biotechnology and Bioprocess Engineering
2011 Volume.16 No. 6 p.1144 ~ p.1151
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Improvement of poly(γ-glutamic acid) biosynthesis and quantitative metabolic flux analysis of a two-stage strategy for agitation speed control in the culture of Bacillus subtilis NX-2
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Zhang Dan
Xu Zongqi
Xu Hong
Feng Xiaohai
Li Sha
Cai Heng
Wei Yan
Ouyang Pingkai
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Abstract
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In this study, the production of poly(γ-glutamic acid) by Bacillus subtilis NX-2 (PGA) at different agitation speeds was investigated. Based on the analysis of specific cell growth rate (μ) and specific PGA formation rate (q p ), a two-stage strategy for agitation speed control was proposed. During the first 24 h, an agitation speed of 600 rpm was used to maintain a high μ for better cell growth, which then reduced to 400 rpm after 24 h to maintain a high q p to enhance PGA production. Using this method, the maximum concentration of PGA reached 40.5 ± 0.91 g/L and the PGA productivity was 0.56 ± 0.012 g/L/h, which was 17.7 and 9.8% higher, respectively, than the best results obtained when a constant agitation speed was used. The flux distributions and the related enzymes of 2-oxoglutarate could be affected by this two-stage strategy for agitation speed. The activity of isocitrate dehydrogenase and glutamate dehydrogenase at the key node of 2-oxoglutarate increased, and more flux distribution was directed to glutamate. The flux distribution from extracellular to intracellular glutamate also increased and improved PGA production as the glutamate uptake rates increased using the agitation-shift control method.
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KEYWORD
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poly(γ-glutamic acid), two-stage control strategy, agitation speed, metabolic flux, enzyme activity
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