Synthesis of Pure <Emphasis Type="Italic">meso</Emphasis>-2,3-Butanediol from Crude Glycerol Using an Engineered Metabolic Pathway in <Emphasis Type="Italic">Escherichia coli</Emphasis> |
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Authors: | Soojin Lee Borim Kim Kyungmoon Park Youngsoon Um Jinwon Lee |
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Institution: | (1) Department of Chemical and Biomolecular Engineering, Sogang University, Seoul, 121-742, South Korea;(2) Department of Biological and Chemical Engineering, Hongik University, Chungcheongnam-do, 339-701, South Korea;(3) Clean Energy Center, Korea Institute of Science and Technology, 39-1 Hawolgok-dong, Seongbuk-gu, Seoul, 136-791, South Korea; |
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Abstract: | meso-2,3-Butanediol (meso-2,3-BDO) is essential for the synthesis of various economically valuable biosynthetic products; however, the production of
meso-2,3-BDO from expensive carbon sources is an obstacle for industrial applications. In this study, genes involved in the synthesis
of 2,3-BDO in Klebsiella pneumoniae were identified and used to genetically modify Escherichia coli for meso-2,3-BDO production. Two 2,3-BDO biosynthesis genes—budA, encoding acetolactate, and meso-budC, encoding meso-SADH—from K. pneumoniae were cloned into the pUC18 plasmid and introduced into E. coli. In 2 l batch culture, the SGSB03 E. coli strain yielded meso-2,3-BDO at 0.31 g/gglucose (with a maximum of 15.7 g/lculture after 48 h) and 0.21 g/gcrude glycerol (with a maximum of 6.9 g/lculture after 48 h). Batch cultures were grown under optimized conditions (aerobic, 6% carbon source, 37 °C, and initial pH 7). To
find the optimal culture conditions for meso-2,3-BDO production, we evaluated the enzyme activity of meso-SADH and the whole cell conversion yield (meso-2,3-BDO/acetoin) of the E. coli SGSB02, which contains pSB02. meso-SADH showed high enzyme activity at 30–37 °C and pH 7 (30.5–41.5 U/mg of protein), and the conversion yield of SGSB02 E. coli was highest at 37–42 °C and a pH of 7 (0.25–0.28 g
meso-2,3-BDO/gacetoin). |
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