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KMID : 0545120120220050659
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Journal of Microbiology and Biotechnology
2012 Volume.22 No. 5 p.659 ~ p.667
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Identification of Factors Regulating Escherichia coli 2,3-Butanediol Production by Continuous Culture and Metabolic Flux Analysis
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Mingshou Lu
Lee Soo-Jin
Kim Bo-Rim
Park Chang-Hun
Oh Min-Kyu
Park Kyung-Moon
Lee Sang-Yup
Lee Jin-Won
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Abstract
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2,3-Butanediol (2,3-BDO) is an organic compound with a wide range of industrial applications. Although Escherichia coli is often used for the production of organic compounds, the wild-type E. coli does not contain two essential genes in the 2,3-BDO biosynthesis pathway, and cannot ferment 2,3-BDO. Therefore, a 2,3-BDO biosynthesis mutant strain of Escherichia coli was constructed and cultured. To determine the optimum culture factors for 2,3-BDO production, experiments were conducted under different culture environments ranging from strongly acidic to neutral pH. The extracellular metabolite profiles were obtained using high-performance liquid chromatography (HPLC), and the intracellular metabolite profiles were analyzed by ultra-performance liquid chromatography and quadruple time-of-flight mass spectrometry (UPLC/ Q-TOF-MS). Metabolic flux analysis (MFA) was used to integrate these profiles. The metabolite profiles showed that 2,3-BDO production favors an acidic environment (pH 5), whereas cell mass favors a neutral environment. Furthermore, when the pH of the culture fell below 5, both the cell growth and 2,3-BDO production were inhibited.
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KEYWORD
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2,3-butanediol fermentation, continuous culture, pH influence, metabolic flux analysis, intracellular metabolic measurements
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