Cargando…

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

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,...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5719400/
https://www.ncbi.nlm.nih.gov/pubmed/29214999
http://dx.doi.org/10.1038/s41598-017-15400-x
_version_ 1783284482018639872
author 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
author_facet 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
author_sort Park, Ye Seop
collection PubMed
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(m) 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).
format Online
Article
Text
id pubmed-5719400
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-57194002017-12-08 Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD(+), for enhancing the production of 3-hydroxypropionic acid 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 Sci Rep Article 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(m) 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). Nature Publishing Group UK 2017-12-07 /pmc/articles/PMC5719400/ /pubmed/29214999 http://dx.doi.org/10.1038/s41598-017-15400-x Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
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
Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD(+), for enhancing the production of 3-hydroxypropionic acid
title Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD(+), for enhancing the production of 3-hydroxypropionic acid
title_full Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD(+), for enhancing the production of 3-hydroxypropionic acid
title_fullStr Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD(+), for enhancing the production of 3-hydroxypropionic acid
title_full_unstemmed Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD(+), for enhancing the production of 3-hydroxypropionic acid
title_short Engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and NAD(+), for enhancing the production of 3-hydroxypropionic acid
title_sort engineering an aldehyde dehydrogenase toward its substrates, 3-hydroxypropanal and nad(+), for enhancing the production of 3-hydroxypropionic acid
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5719400/
https://www.ncbi.nlm.nih.gov/pubmed/29214999
http://dx.doi.org/10.1038/s41598-017-15400-x
work_keys_str_mv AT parkyeseop engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT choiunjong engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT namnguyenhoai engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT choisangjin engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT nasirabdul engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT leesungu engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT kimkyungjin engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT junggyooyeol engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT choisangdun engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT shimjeungyeop engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT parksunghoon engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid
AT yootaehyeon engineeringanaldehydedehydrogenasetowarditssubstrates3hydroxypropanalandnadforenhancingtheproductionof3hydroxypropionicacid