Phage-Assisted Evolution of Bacillus methanolicus Methanol Dehydrogenase 2.
Alcohol Oxidoreductases
/ genetics
Bacillus
/ genetics
Bacterial Proteins
/ genetics
Bacteriophages
/ genetics
Escherichia coli
/ genetics
Formaldehyde
/ metabolism
Gene Expression Regulation, Bacterial
/ genetics
Metabolic Engineering
/ methods
Metabolic Networks and Pathways
/ genetics
Methanol
NAD
/ genetics
directed evolution
methanol assimilation
methanol dehydrogenase
phage-assisted continuous evolution
phage-assisted noncontinuous evolution
synthetic methylotrophy
Journal
ACS synthetic biology
ISSN: 2161-5063
Titre abrégé: ACS Synth Biol
Pays: United States
ID NLM: 101575075
Informations de publication
Date de publication:
19 04 2019
19 04 2019
Historique:
pubmed:
12
3
2019
medline:
4
3
2020
entrez:
12
3
2019
Statut:
ppublish
Résumé
Synthetic methylotrophy, the modification of organisms such as E. coli to grow on methanol, is a longstanding goal of metabolic engineering and synthetic biology. The poor kinetic properties of NAD-dependent methanol dehydrogenase, the first enzyme in most methanol assimilation pathways, limit pathway flux and present a formidable challenge to synthetic methylotrophy. To address this bottleneck, we used a formaldehyde biosensor to develop a phage-assisted noncontinuous evolution (PANCE) selection for variants of Bacillus methanolicus methanol dehydrogenase 2 (Bm Mdh2). Using this selection, we evolved Mdh2 variants with up to 3.5-fold improved V
Identifiants
pubmed: 30856338
doi: 10.1021/acssynbio.8b00481
pmc: PMC6479731
doi:
Substances chimiques
Bacterial Proteins
0
NAD
0U46U6E8UK
Formaldehyde
1HG84L3525
Alcohol Oxidoreductases
EC 1.1.-
alcohol dehydrogenase (acceptor)
EC 1.1.2.8
Methanol
Y4S76JWI15
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
796-806Subventions
Organisme : NIBIB NIH HHS
ID : R01 EB022376
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM118062
Pays : United States
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