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Lactobacillus reuteri NAD(P)H oxidase : Properties and coexpression with propanediol-utilization enzymes for enhancing 3-hydroxypropionic acid production from 3-hydroxypropionaldehyde

Dishisha, Tarek LU ; Sabet-Azad, Ramin LU ; Arieta, Victor and Hatti-Kaul, Rajni LU (2019) In Journal of Biotechnology 289. p.135-143
Abstract

Lactobacillus reuteri metabolizes glycerol through propanediol-utilization (Pdu) pathway to 1,3-propanediol (1,3-PD) via 3-hydroxypropionaldehyde (3-HPA) as intermediate. In the resting cells, the oxidized co-factor obtained in the reaction is regenerated by simultaneous oxidation of 3-HPA to 3-hydroxypropionic acid (3-HP) using propionaldehyde dehydrogenase (PduP), phosphotransacylase (PduL) and propionate kinase (PduW). We have earlier shown that the use of resting cells of recombinant Escherichia coli expressing the oxidative pathway gives the highest theoretical yield of 1 mol 3-HP per mol 3-HPA but is limited by cofactor depletion. In the present study, the gene encoding the enzyme NAD(P)H oxidase (LreuNox) that utilizes molecular... (More)

Lactobacillus reuteri metabolizes glycerol through propanediol-utilization (Pdu) pathway to 1,3-propanediol (1,3-PD) via 3-hydroxypropionaldehyde (3-HPA) as intermediate. In the resting cells, the oxidized co-factor obtained in the reaction is regenerated by simultaneous oxidation of 3-HPA to 3-hydroxypropionic acid (3-HP) using propionaldehyde dehydrogenase (PduP), phosphotransacylase (PduL) and propionate kinase (PduW). We have earlier shown that the use of resting cells of recombinant Escherichia coli expressing the oxidative pathway gives the highest theoretical yield of 1 mol 3-HP per mol 3-HPA but is limited by cofactor depletion. In the present study, the gene encoding the enzyme NAD(P)H oxidase (LreuNox) that utilizes molecular oxygen as substrate, was isolated from L. reuteri and heterologously overexpressed in E. coli. LreuNox has a pH optimum of 6 and exhibits Vmax of 101.1 ± 2.2 U/mg with NADH, which is 30% higher than that for NADPH. Co-expression of LreuNox with PduP, PduL and PduW in E. coli enhances the biocatalytic lifetime as well as productivity at least two-fold compared to that achieved without co-factor regeneration.

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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
3-Hydroxypropionaldehyde, 3-Hydroxypropionic acid, Co-factor regeneration, Fed-batch biotransformation, Lactobacillus reuteri, NADH oxidase
in
Journal of Biotechnology
volume
289
pages
9 pages
publisher
Elsevier
external identifiers
  • scopus:85057577510
  • pmid:30503904
ISSN
0168-1656
DOI
10.1016/j.jbiotec.2018.11.010
language
English
LU publication?
yes
id
67927e08-2047-4397-a641-338b4fee6f03
date added to LUP
2018-12-18 08:36:03
date last changed
2024-04-15 19:13:53
@article{67927e08-2047-4397-a641-338b4fee6f03,
  abstract     = {{<p>Lactobacillus reuteri metabolizes glycerol through propanediol-utilization (Pdu) pathway to 1,3-propanediol (1,3-PD) via 3-hydroxypropionaldehyde (3-HPA) as intermediate. In the resting cells, the oxidized co-factor obtained in the reaction is regenerated by simultaneous oxidation of 3-HPA to 3-hydroxypropionic acid (3-HP) using propionaldehyde dehydrogenase (PduP), phosphotransacylase (PduL) and propionate kinase (PduW). We have earlier shown that the use of resting cells of recombinant Escherichia coli expressing the oxidative pathway gives the highest theoretical yield of 1 mol 3-HP per mol 3-HPA but is limited by cofactor depletion. In the present study, the gene encoding the enzyme NAD(P)H oxidase (LreuNox) that utilizes molecular oxygen as substrate, was isolated from L. reuteri and heterologously overexpressed in E. coli. LreuNox has a pH optimum of 6 and exhibits V<sub>max</sub> of 101.1 ± 2.2 U/mg with NADH, which is 30% higher than that for NADPH. Co-expression of LreuNox with PduP, PduL and PduW in E. coli enhances the biocatalytic lifetime as well as productivity at least two-fold compared to that achieved without co-factor regeneration.</p>}},
  author       = {{Dishisha, Tarek and Sabet-Azad, Ramin and Arieta, Victor and Hatti-Kaul, Rajni}},
  issn         = {{0168-1656}},
  keywords     = {{3-Hydroxypropionaldehyde; 3-Hydroxypropionic acid; Co-factor regeneration; Fed-batch biotransformation; Lactobacillus reuteri; NADH oxidase}},
  language     = {{eng}},
  pages        = {{135--143}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Biotechnology}},
  title        = {{Lactobacillus reuteri NAD(P)H oxidase : Properties and coexpression with propanediol-utilization enzymes for enhancing 3-hydroxypropionic acid production from 3-hydroxypropionaldehyde}},
  url          = {{http://dx.doi.org/10.1016/j.jbiotec.2018.11.010}},
  doi          = {{10.1016/j.jbiotec.2018.11.010}},
  volume       = {{289}},
  year         = {{2019}},
}