PMID- 28479191
OWN - NLM
STAT- MEDLINE
DCOM- 20180409
LR  - 20220331
IS  - 1096-7184 (Electronic)
IS  - 1096-7176 (Print)
IS  - 1096-7176 (Linking)
VI  - 42
DP  - 2017 Jul
TI  - Isotopically nonstationary (13)C flux analysis of cyanobacterial isobutyraldehyde 
      production.
PG  - 9-18
LID - S1096-7176(16)30156-2 [pii]
LID - 10.1016/j.ymben.2017.05.001 [doi]
AB  - We applied isotopically nonstationary (13)C metabolic flux analysis (INST-MFA) to 
      compare the pathway fluxes of wild-type (WT) Synechococcus elongatus PCC 7942 to 
      an engineered strain (SA590) that produces isobutyraldehyde (IBA). The flux maps 
      revealed a potential bottleneck at the pyruvate kinase (PK) reaction step that 
      was associated with diversion of flux into a three-step PK bypass pathway 
      involving the enzymes PEP carboxylase (PEPC), malate dehydrogenase (MDH), and 
      malic enzyme (ME). Overexpression of pk in SA590 led to a significant improvement 
      in IBA specific productivity. Single-gene overexpression of the three enzymes in 
      the proposed PK bypass pathway also led to improvements in IBA production, 
      although to a lesser extent than pk overexpression. Combinatorial overexpression 
      of two of the three genes in the proposed PK bypass pathway (mdh and me) led to 
      improvements in specific productivity that were similar to those achieved by 
      single-gene pk overexpression. Our work demonstrates how (13)C flux analysis can 
      be used to identify potential metabolic bottlenecks and novel metabolic routes, 
      and how these findings can guide rational metabolic engineering of cyanobacteria 
      for increased production of desired molecules.
CI  - Copyright (c) 2017 International Metabolic Engineering Society. Published by 
      Elsevier Inc. All rights reserved.
FAU - Jazmin, Lara J
AU  - Jazmin LJ
AD  - Department of Chemical and Biomolecular Engineering, Vanderbilt University, 
      Nashville, TN 37235, USA.
FAU - Xu, Yao
AU  - Xu Y
AD  - Department of Biological Sciences, Vanderbilt University, Nashville, TN 27235, 
      USA.
FAU - Cheah, Yi Ern
AU  - Cheah YE
AD  - Department of Chemical and Biomolecular Engineering, Vanderbilt University, 
      Nashville, TN 37235, USA.
FAU - Adebiyi, Adeola O
AU  - Adebiyi AO
AD  - Department of Chemical and Biomolecular Engineering, Vanderbilt University, 
      Nashville, TN 37235, USA.
FAU - Johnson, Carl Hirschie
AU  - Johnson CH
AD  - Department of Biological Sciences, Vanderbilt University, Nashville, TN 27235, 
      USA; Department of Molecular Physiology and Biophysics, Vanderbilt University, 
      Nashville, TN 27235, USA.
FAU - Young, Jamey D
AU  - Young JD
AD  - Department of Chemical and Biomolecular Engineering, Vanderbilt University, 
      Nashville, TN 37235, USA; Department of Molecular Physiology and Biophysics, 
      Vanderbilt University, Nashville, TN 27235, USA. Electronic address: 
      j.d.young@vanderbilt.edu.
LA  - eng
GR  - R01 GM067152/GM/NIGMS NIH HHS/United States
GR  - R01 GM107434/GM/NIGMS NIH HHS/United States
GR  - R37 GM067152/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20170504
PL  - Belgium
TA  - Metab Eng
JT  - Metabolic engineering
JID - 9815657
RN  - 0 (Aldehydes)
RN  - 0 (Bacterial Proteins)
RN  - 0 (Carbon Isotopes)
RN  - C42E28168L (isobutyraldehyde)
RN  - EC 1.1.1.37 (Malate Dehydrogenase)
RN  - EC 2.7.1.40 (Pyruvate Kinase)
RN  - EC 4.1.1.31 (Phosphoenolpyruvate Carboxylase)
SB  - IM
MH  - Aldehydes/*metabolism
MH  - Bacterial Proteins/genetics/metabolism
MH  - Carbon Isotopes/*metabolism
MH  - Malate Dehydrogenase/genetics/metabolism
MH  - Phosphoenolpyruvate Carboxylase/genetics/metabolism
MH  - Pyruvate Kinase/genetics/metabolism
MH  - Staining and Labeling/*methods
MH  - Synechococcus/genetics/*metabolism
PMC - PMC5660605
MID - NIHMS911674
OTO - NOTNLM
OT  - (13)C INST-MFA
OT  - Isobutyraldehyde
OT  - Nonstationary (13)C metabolic flux analysis
OT  - Photoautotrophic metabolism
OT  - Pyruvate kinase
OT  - Synechococcus elongatus PCC7942
EDAT- 2017/05/10 06:00
MHDA- 2018/04/10 06:00
PMCR- 2018/01/01
CRDT- 2017/05/09 06:00
PHST- 2016/10/06 00:00 [received]
PHST- 2017/04/11 00:00 [revised]
PHST- 2017/05/03 00:00 [accepted]
PHST- 2017/05/10 06:00 [pubmed]
PHST- 2018/04/10 06:00 [medline]
PHST- 2017/05/09 06:00 [entrez]
PHST- 2018/01/01 00:00 [pmc-release]
AID - S1096-7176(16)30156-2 [pii]
AID - 10.1016/j.ymben.2017.05.001 [doi]
PST - ppublish
SO  - Metab Eng. 2017 Jul;42:9-18. doi: 10.1016/j.ymben.2017.05.001. Epub 2017 May 4.