Lactobacillus reuteri NAD(P)H oxidase : Properties and coexpression with propanediol-utilization enzymes for enhancing 3-hydroxypropionic acid production from 3-hydroxypropionaldehyde
(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.
(Less)
- author
- Dishisha, Tarek LU ; Sabet-Azad, Ramin LU ; Arieta, Victor and Hatti-Kaul, Rajni LU
- organization
- publishing date
- 2019
- 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
- 2024-09-17 10:14:33
@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}}, }