Abstract
Klebsiella pneumoniae was engineered to produce 2-butanol from crude glycerol as a sole carbon source by expressing acetolactate synthase (ilvIH), keto-acid reducto-isomerase (ilvC) and dihydroxy-acid dehydratase (ilvD) from K. pneumoniae, and α-ketoisovalerate decarboxylase (kivd) and alcohol dehydrogenase (adhA) from Lactococcus lactis. Engineered K. pneumonia, ∆ldhA/pBR-iBO (ilvIH–ilvC–ilvD–kivd–adhA), produced 2-butanol (160 mg l−1) from crude glycerol. To increase the yield of 2-butanol, we eliminated the 2,3-butanediol pathway from the recombinant strain by inactivating α-acetolactate decarboxylase (adc). This further engineering step improved the yield of 2-butanol from 160 to 320 mg l−1. This represents the first successful attempt to produce 2-butanol from crude glycerol.
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This work was supported by Korea Ministry of Environment and the Joint Degree and Research Center funded by Korea Research Council of Fundamental Science & Technology.
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(A) Construction of an acetolactate decarboxylase-deficient mutant of K. pneumoniae ΔldhA strain by replacing adc with an apramycin-resistance gene [aac(3)IV] through homologous recombination. (B) Correct recombination was confirmed by Southern blot hybridization of PstI-digested chromosomal DNA using the probes Apr (apramycin-resistance gene) and adc (DNA fragment upstream of adc) (DOC 33 kb)
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Oh, BR., Heo, SY., Lee, SM. et al. Production of 2-butanol from crude glycerol by a genetically-engineered Klebsiella pneumoniae strain. Biotechnol Lett 36, 57–62 (2014). https://doi.org/10.1007/s10529-013-1333-0
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DOI: https://doi.org/10.1007/s10529-013-1333-0