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).
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
- Aldehyde dehydrogenase
- 3-Hydroxypropionic acid
- Klebsiella pneumoniae
- Substrate specificity