Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD, for enhancing the production of 3-hydroxypropionic acid.

Citation data:

Scientific reports, ISSN: 2045-2322, Vol: 7, Issue: 1, Page: 17155

Publication Year:
2017
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Repository URL:
http://scholarworks.unist.ac.kr/handle/201301/24273
PMID:
29214999
DOI:
10.1038/s41598-017-15400-x
Author(s):
Park, Ye Seop; Choi, Un Jong; Nam, Nguyen Hoai; Choi, Sang Jin; Nasir, Abdul; Lee, Sun-Gu; Kim, Kyung Jin; Jung, Gyoo Yeol; Choi, Sangdun; Shim, Jeung Yeop; Park, Sunghoon; Yoo, Tae Hyeon Show More Hide
Publisher(s):
Springer Nature; NATURE PUBLISHING GROUP
Tags:
Multidisciplinary
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article description
3-Hydroxypropionic acid (3-HP) can be produced via the biological route involving two enzymatic reactions: dehydration of glycerol to 3-hydroxypropanal (3-HPA) and then oxidation to 3-HP. However, commercial production of 3-HP using recombinant microorganisms has been hampered with several problems, some of which are associated with the toxicity of 3-HPA and the efficiency of NAD regeneration. We engineered α-ketoglutaric semialdehyde dehydrogenase (KGSADH) from Azospirillum brasilense for the second reaction to address these issues. The residues in the binding sites for the substrates, 3-HPA and NAD, were randomized, and the resulting libraries were screened for higher activity. Isolated KGSADH variants had significantly lower K values for both the substrates. The enzymes also showed higher substrate specificities for aldehyde and NAD, less inhibition by NADH, and greater resistance to inactivation by 3-HPA than the wild-type enzyme. A recombinant Pseudomonas denitrificans strain with one of the engineered KGSADH variants exhibited less accumulation of 3-HPA, decreased levels of inactivation of the enzymes, and higher cell growth than that with the wild-type KGSADH. The flask culture of the P. denitrificans strain with the mutant KGSADH resulted in about 40% increase of 3-HP titer (53 mM) compared with that using the wild-type enzyme (37 mM).