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3-Hydroxypropionaldehyde-specific aldehyde dehydrogenase from Bacillus subtilis catalyzes 3-hydroxypropionic acid production in Klebsiella pneumoniae

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In Klebsiella pneumoniae, aldehyde dehydrogenases (ALDH) convert 3-hydroxypropionaldehyde (3-HPA) into 3-hydroxypropionic acid (3-HP). Although ALDHs can increase the production of 3-HP in K. pneumoniae, the substrate specificity of ALDH homologues from other microorganisms toward 3-HPA is less documented. Here we report that DhaS, a putative ALDH from Bacillus subtilis, shows high specificity toward 3-HPA when heterologously expressed in K. pneumoniae. Using NAD+ as a cofactor, DhaS exhibited higher catalytic activity (2.3 U mg−1) and lower K m value (0.4 mmol l−1) toward 3-HPA than that toward other aldehydes. Under shake-flask conditions, the recombinant strain produced 2.1 g 3-HP l−1 in 24 h, which is 3.9-fold of that in a control harboring a blank vector. Under non-optimized bioreactor conditions, the recombinant strain produced 18 g 3-HP l−1 and 1,3-propanediol (1,3-PDO) at 27 g l−1 in 24 h. The overall conversion rate from glycerol to 3-HP and 1,3-PDO reached 59.4 mol mol−1. Homology modeling of DhaS illustrates substrate specificity and NAD+-binding site. DhaS is thus a 3-HPA-specific enzyme useful for production of 3-HP.

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This work was supported by Grants from National Basic Research Program of China (973 Program) (2012CB725200) and National Natural Science Foundation of China (No. 21076013, 21276014).

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Correspondence to Pingfang Tian.

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Mingyue Su and Ying Li have contributed equally to this work.

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Su, M., Li, Y., Ge, X. et al. 3-Hydroxypropionaldehyde-specific aldehyde dehydrogenase from Bacillus subtilis catalyzes 3-hydroxypropionic acid production in Klebsiella pneumoniae . Biotechnol Lett 37, 717–724 (2015). https://doi.org/10.1007/s10529-014-1730-z

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  • Aldehyde dehydrogenase
  • Glycerol
  • 3-Hydroxypropionaldehyde
  • 3-Hydroxypropionic acid
  • Klebsiella pneumoniae
  • Substrate specificity