PMID- 34970474
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220429
IS  - 2197-4365 (Print)
IS  - 2197-4365 (Electronic)
IS  - 2197-4365 (Linking)
VI  - 8
IP  - 1
DP  - 2021
TI  - Strain engineering and bioprocessing strategies for biobased production of 
      porphobilinogen in Escherichia coli.
PG  - 122
LID - 10.1186/s40643-021-00482-3 [doi]
LID - 122
AB  - Strain engineering and bioprocessing strategies were applied for biobased 
      production of porphobilinogen (PBG) using Escherichia coli as the cell factory. 
      The non-native Shemin/C4 pathway was first implemented by heterologous expression 
      of hemA from Rhodopseudomonas spheroids to supply carbon flux from the natural 
      tricarboxylic acid (TCA) pathways for PBG biosynthesis via succinyl-CoA. 
      Metabolic strategies were then applied for carbon flux direction from the TCA 
      pathways to the C4 pathway. To promote PBG stability and accumulation, Clustered 
      Regularly Interspersed Short Palindromic Repeats interference (CRISPRi) was 
      applied to repress hemC expression and, therefore, reduce carbon flowthrough 
      toward porphyrin biosynthesis with minimal impact to cell physiology. To further 
      enhance PBG biosynthesis and accumulation under the hemC-repressed genetic 
      background, we further heterologously expressed native E. coli hemB. Using these 
      engineered E. coli strains for bioreactor cultivation based on ~ 30 g L(-1) 
      glycerol, we achieved high PBG titers up to 209 mg L(-1), representing 1.73% of 
      the theoretical PBG yield, with improved PBG stability and accumulation. 
      Potential biochemical, genetic, and metabolic factors limiting PBG production 
      were systematically identified for characterization. SUPPLEMENTARY INFORMATION: 
      The online version contains supplementary material available at 
      10.1186/s40643-021-00482-3.
CI  - (c) The Author(s) 2021.
FAU - Lall, Davinder
AU  - Lall D
AD  - Department of Chemical Engineering, University of Waterloo, 200 University Avenue 
      West, Waterloo, ON N2L 3G1 Canada. GRID: grid.46078.3d. ISNI: 0000 0000 8644 1405
FAU - Miscevic, Dragan
AU  - Miscevic D
AD  - Department of Chemical Engineering, University of Waterloo, 200 University Avenue 
      West, Waterloo, ON N2L 3G1 Canada. GRID: grid.46078.3d. ISNI: 0000 0000 8644 1405
FAU - Bruder, Mark
AU  - Bruder M
AD  - Department of Chemical Engineering, University of Waterloo, 200 University Avenue 
      West, Waterloo, ON N2L 3G1 Canada. GRID: grid.46078.3d. ISNI: 0000 0000 8644 1405
FAU - Westbrook, Adam
AU  - Westbrook A
AD  - Department of Chemical Engineering, University of Waterloo, 200 University Avenue 
      West, Waterloo, ON N2L 3G1 Canada. GRID: grid.46078.3d. ISNI: 0000 0000 8644 1405
FAU - Aucoin, Marc
AU  - Aucoin M
AD  - Department of Chemical Engineering, University of Waterloo, 200 University Avenue 
      West, Waterloo, ON N2L 3G1 Canada. GRID: grid.46078.3d. ISNI: 0000 0000 8644 1405
FAU - Moo-Young, Murray
AU  - Moo-Young M
AD  - Department of Chemical Engineering, University of Waterloo, 200 University Avenue 
      West, Waterloo, ON N2L 3G1 Canada. GRID: grid.46078.3d. ISNI: 0000 0000 8644 1405
FAU - Perry Chou, C
AU  - Perry Chou C
AUID- ORCID: 0000-0002-7254-8118
AD  - Department of Chemical Engineering, University of Waterloo, 200 University Avenue 
      West, Waterloo, ON N2L 3G1 Canada. GRID: grid.46078.3d. ISNI: 0000 0000 8644 1405
LA  - eng
PT  - Journal Article
DEP - 20211213
PL  - Germany
TA  - Bioresour Bioprocess
JT  - Bioresources and bioprocessing
JID - 101665551
PMC - PMC8668860
OTO - NOTNLM
OT  - Escherichia coli
OT  - Glycerol
OT  - Glyoxylate shunt
OT  - Porphobilinogen (PBG)
OT  - Strain engineering
OT  - Succinyl-CoA
OT  - Tricarboxylic acid (TCA) cycle
COIS- Competing interestsThe authors declare that they have no competing interests.
EDAT- 2022/01/01 06:00
MHDA- 2022/01/01 06:01
PMCR- 2021/12/13
CRDT- 2021/12/31 06:05
PHST- 2021/09/03 00:00 [received]
PHST- 2021/12/04 00:00 [accepted]
PHST- 2021/12/31 06:05 [entrez]
PHST- 2022/01/01 06:00 [pubmed]
PHST- 2022/01/01 06:01 [medline]
PHST- 2021/12/13 00:00 [pmc-release]
AID - 482 [pii]
AID - 10.1186/s40643-021-00482-3 [doi]
PST - ppublish
SO  - Bioresour Bioprocess. 2021;8(1):122. doi: 10.1186/s40643-021-00482-3. Epub 2021 
      Dec 13.