Search Menu
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information
Search Scholarly Publications

Title: Complete enzyme set for chlorophyll biosynthesis in Escherichia coli

Abstract

Chlorophylls are essential cofactors for photosynthesis, which sustains global food chains and oxygen production. Billions of tons of chlorophylls are synthesized annually, yet full understanding of chlorophyll biosynthesis has been hindered by the lack of characterization of the Mg–protoporphyrin IX monomethyl ester oxidative cyclase step, which confers the distinctive green color of these pigments. We demonstrate cyclase activity using heterologously expressed enzyme. Next, we assemble a genetic module that encodes the complete chlorophyll biosynthetic pathway and show that it functions in Escherichia coli. Expression of 12 genes converts endogenous protoporphyrin IX into chlorophyll a, turning E. coli cells green. Our results delineate a minimum set of enzymes required to make chlorophyll and establish a platform for engineering photosynthesis in a heterotrophic model organism.

Authors:
ORCiD logo [1];  [1]; ORCiD logo [1];  [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [1]
+ Show Author Affiliations
  1. Univ. of Sheffield (United Kingdom)
  2. Pennsylvania State Univ., University Park, PA (United States)
Publication Date:
Research Org.:
Pennsylvania State Univ., University Park, PA (United States); Washington Univ., St. Louis, MO (United States); Energy Frontier Research Centers (EFRC) (United States). Photosynthetic Antenna Research Center (PARC)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences, and Biosciences Division
OSTI Identifier:
1499942
Grant/Contract Number:  
FG02-94ER20137; SC0001035
Resource Type:
Accepted Manuscript
Journal Name:
Science Advances
Additional Journal Information:
Journal Volume: 4; Journal Issue: 1; Journal ID: ISSN 2375-2548
Publisher:
AAAS
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Chen, Guangyu E., Canniffe, Daniel P., Barnett, Samuel F. H., Hollingshead, Sarah, Brindley, Amanda A., Vasilev, Cvetelin, Bryant, Donald A., and Hunter, C. Neil. Complete enzyme set for chlorophyll biosynthesis in Escherichia coli. United States: N. p., 2018. Web. doi:10.1126/sciadv.aaq1407.
Copy to clipboard
Chen, Guangyu E., Canniffe, Daniel P., Barnett, Samuel F. H., Hollingshead, Sarah, Brindley, Amanda A., Vasilev, Cvetelin, Bryant, Donald A., & Hunter, C. Neil. Complete enzyme set for chlorophyll biosynthesis in Escherichia coli. United States. https://doi.org/10.1126/sciadv.aaq1407
Copy to clipboard
Chen, Guangyu E., Canniffe, Daniel P., Barnett, Samuel F. H., Hollingshead, Sarah, Brindley, Amanda A., Vasilev, Cvetelin, Bryant, Donald A., and Hunter, C. Neil. Fri . "Complete enzyme set for chlorophyll biosynthesis in Escherichia coli". United States. https://doi.org/10.1126/sciadv.aaq1407. https://www.osti.gov/servlets/purl/1499942.
Copy to clipboard
@article{osti_1499942,
title = {Complete enzyme set for chlorophyll biosynthesis in Escherichia coli},
author = {Chen, Guangyu E. and Canniffe, Daniel P. and Barnett, Samuel F. H. and Hollingshead, Sarah and Brindley, Amanda A. and Vasilev, Cvetelin and Bryant, Donald A. and Hunter, C. Neil},
abstractNote = {Chlorophylls are essential cofactors for photosynthesis, which sustains global food chains and oxygen production. Billions of tons of chlorophylls are synthesized annually, yet full understanding of chlorophyll biosynthesis has been hindered by the lack of characterization of the Mg–protoporphyrin IX monomethyl ester oxidative cyclase step, which confers the distinctive green color of these pigments. We demonstrate cyclase activity using heterologously expressed enzyme. Next, we assemble a genetic module that encodes the complete chlorophyll biosynthetic pathway and show that it functions in Escherichia coli. Expression of 12 genes converts endogenous protoporphyrin IX into chlorophyll a, turning E. coli cells green. Our results delineate a minimum set of enzymes required to make chlorophyll and establish a platform for engineering photosynthesis in a heterotrophic model organism.},
doi = {10.1126/sciadv.aaq1407},
journal = {Science Advances},
number = 1,
volume = 4,
place = {United States},
year = {2018},
month = {1}
}
Copy to clipboard
Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1126/sciadv.aaq1407
Copyright Statement
Other availability
Search WorldCat Search WorldCat to find libraries that may hold this journal
Citation Metrics:
Cited by: 26 works
Citation information provided by
Web of Science

Figures / Tables:

Fig. 1. Fig. 1.: Assembly of the Chl biosynthesis pathway in Escherichia coli. (A) Overall reactions from PPIX to Chl a catalyzed by the enzymes introduced to E. coli. The insertion of Fe2+ into PPIX (not shown) creates a biosynthetic branch point (not shown) that yields heme. Colored shading denotes the chemicalmore » change(s) at each step. ATP, adenosine triphosphate; ADP, adenosine diphosphate; SAM, S-adenosine-L-methionine; SAH, S-adenosyl-L-homocysteine; NADP+, nicotinamide adenine dinucleotide phosphate; NADPH, reduced form of NADP+; Pi, inorganic phosphate; PPi, inorganic pyrophosphate. (B) Arrangement and relative size of each gene in the constructed plasmids. The chlI, chlD, chlH, gun4, chlM, acsF, dvr (bciB), and chlG genes were consecutively cloned into a modified pET3a vector with a single T7 promoter upstream of chlI and a ribosome-binding site upstream of each gene using the link-and-lock method (see fig. S2 for details) (40). Colors for genes correspond to those used in (A). Plasmid constructs and gene contents are IM (chlI–chlM), IA (chlI–acsF), ID (chlI–dvr), IG (chlI–chlG), BoP (BoWSCP and chlP), and DE (dxs and crtE). BoP is a pACYCDuet1-based plasmid containing a sequence encoding the BoWSCP protein with a C-terminal His10tag (10) and the Synechocystis chlP gene. DE is a pCOLADuet1-based plasmid containing the E. coli dxs gene and the Rvi. gelatinosus crtE gene.« less
All figures and tables (4 total)
Save / Share:
Export Metadata
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.

Works referenced in this record:

A Cyanobacterial Chlorophyll Synthase-HliD Complex Associates with the Ycf39 Protein and the YidC/Alb3 Insertase
journal, March 2014

  • Chidgey, Jack W.; Linhartová, Markéta; Komenda, Josef
  • The Plant Cell, Vol. 26, Issue 3
  • DOI: 10.1105/tpc.114.124495

A Cyanobacterial Chlorophyll Synthase-HliD Complex Associates with the Ycf39 Protein and the YidC/Alb3 Insertase
journal, March 2014

  • Chidgey, Jack W.; Linhartová, Markéta; Komenda, Josef
  • The Plant Cell, Vol. 26, Issue 3
  • DOI: 10.1105/tpc.114.124495

BioBrickTM compatible vector system for protein expression in Rhodobacter sphaeroides
journal, February 2014

  • Tikh, Ilya B.; Held, Mark; Schmidt-Dannert, Claudia
  • Applied Microbiology and Biotechnology, Vol. 98, Issue 7
  • DOI: 10.1007/s00253-014-5527-8

Detection and Partial Characterization of Activity of Chlorophyll Synthetase in Etioplast Membranes
journal, August 1980

Differential requirement of two homologous proteins encoded by sll1214 and sll1874 for the reaction of Mg protoporphyrin monomethylester oxidative cyclase under aerobic and micro-oxic growth conditions
journal, December 2009

  • Peter, Enrico; Salinas, Annabel; Wallner, Thomas
  • Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1787, Issue 12
  • DOI: 10.1016/j.bbabio.2009.06.006

Molecular mechanism of photosystem I assembly in oxygenic organisms
journal, September 2015

  • Yang, Huixia; Liu, Jun; Wen, Xiaogang
  • Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1847, Issue 9
  • DOI: 10.1016/j.bbabio.2014.12.011

Three-Dimensional Resolution Doubling in Wide-Field Fluorescence Microscopy by Structured Illumination
journal, June 2008

Bacterial Reaction Centers Purified with Styrene Maleic Acid Copolymer Retain Native Membrane Functional Properties and Display Enhanced Stability
journal, September 2014

  • Swainsbury, David J. K.; Scheidelaar, Stefan; van Grondelle, Rienk
  • Angewandte Chemie International Edition, Vol. 53, Issue 44
  • DOI: 10.1002/anie.201406412

Detection and Partial Characterization of Activity of Chlorophyll Synthetase in Etioplast Membranes
journal, August 1980

The biosynthesis of adenosylcobalamin (vitamin B12)
journal, June 2002

  • Warren, Martin J.; Raux, Evelyne; Schubert, Heidi L.
  • Natural Product Reports, Vol. 19, Issue 4
  • DOI: 10.1039/b108967f

Three classes of oxygen-dependent cyclase involved in chlorophyll and bacteriochlorophyll biosynthesis
journal, May 2017

  • Chen, Guangyu E.; Canniffe, Daniel P.; Hunter, C. Neil
  • Proceedings of the National Academy of Sciences, Vol. 114, Issue 24
  • DOI: 10.1073/pnas.1701687114

Elucidation of the biosynthesis of the methane catalyst coenzyme F430
journal, February 2017

  • Moore, Simon J.; Sowa, Sven T.; Schuchardt, Christopher
  • Nature, Vol. 543, Issue 7643
  • DOI: 10.1038/nature21427

Generic Assignments, Strain Histories and Properties of Pure Cultures of Cyanobacteria
journal, March 1979

  • Stanier, Roger Y.; Deruelles, Josette; Rippka, Rosmarie
  • Microbiology, Vol. 111, Issue 1, p. 1-61
  • DOI: 10.1099/00221287-111-1-1

A Broad Host Range Mobilization System for In Vivo Genetic Engineering Transposon Mutagenesis in Gram Negative Bacteria
journal, November 1983

  • Simon, R.; Priefer, U.; Pühler, A.
  • Bio/Technology, Vol. 1, Issue 9, p. 784-791
  • DOI: 10.1038/nbt1183-784

BioBrickTM compatible vector system for protein expression in Rhodobacter sphaeroides
journal, February 2014

  • Tikh, Ilya B.; Held, Mark; Schmidt-Dannert, Claudia
  • Applied Microbiology and Biotechnology, Vol. 98, Issue 7
  • DOI: 10.1007/s00253-014-5527-8

Xantha-l encodes a membrane subunit of the aerobic Mg-protoporphyrin IX monomethyl ester cyclase involved in chlorophyll biosynthesis
journal, April 2005

  • Rzeznicka, K.; Walker, C. J.; Westergren, T.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 16
  • DOI: 10.1073/pnas.0501784102

Synthesis of Chlorophyll-Binding Proteins in a Fully Segregated Δycf54 Strain of the Cyanobacterium Synechocystis PCC 6803
journal, March 2016

  • Hollingshead, Sarah; Kopečná, Jana; Armstrong, David R.
  • Frontiers in Plant Science, Vol. 7
  • DOI: 10.3389/fpls.2016.00292

Photosystem II Assembly: From Cyanobacteria to Plants
journal, April 2013

Anaerobic chlorophyll isocyclic ring formation in Rhodobacter capsulatus requires a cobalamin cofactor
journal, June 2000

  • Gough, S. P.; Petersen, B. O.; Duus, J. O.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 12
  • DOI: 10.1073/pnas.97.12.6908

The biochemistry of heme biosynthesis
journal, June 2008

  • Heinemann, Ilka U.; Jahn, Martina; Jahn, Dieter
  • Archives of Biochemistry and Biophysics, Vol. 474, Issue 2
  • DOI: 10.1016/j.abb.2008.02.015

The biosynthetic pathway of coenzyme F430 in methanogenic and methanotrophic archaea
journal, October 2016

Biosynthesis of Chlorophyll a in a Purple Bacterial Phototroph and Assembly into a Plant Chlorophyll–Protein Complex
journal, May 2016

  • Hitchcock, Andrew; Jackson, Philip J.; Chidgey, Jack W.
  • ACS Synthetic Biology, Vol. 5, Issue 9
  • DOI: 10.1021/acssynbio.6b00069

The biosynthesis of adenosylcobalamin (vitamin B12)
journal, June 2002

  • Warren, Martin J.; Raux, Evelyne; Schubert, Heidi L.
  • Natural Product Reports, Vol. 19, Issue 4
  • DOI: 10.1039/b108967f

Xantha-l encodes a membrane subunit of the aerobic Mg-protoporphyrin IX monomethyl ester cyclase involved in chlorophyll biosynthesis
journal, April 2005

  • Rzeznicka, K.; Walker, C. J.; Westergren, T.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 16
  • DOI: 10.1073/pnas.0501784102

Conserved Chloroplast Open-reading Frame ycf54 Is Required for Activity of the Magnesium Protoporphyrin Monomethylester Oxidative Cyclase in Synechocystis PCC 6803
journal, June 2012

  • Hollingshead, Sarah; Kopečná, Jana; Jackson, Philip J.
  • Journal of Biological Chemistry, Vol. 287, Issue 33
  • DOI: 10.1074/jbc.m112.352526

Structure and function of enzymes in heme biosynthesis
journal, May 2010

  • Layer, Gunhild; Reichelt, Joachim; Jahn, Dieter
  • Protein Science, Vol. 19, Issue 6
  • DOI: 10.1002/pro.405

Editorial: Assembly of the Photosystem II Membrane-Protein Complex of Oxygenic Photosynthesis
journal, May 2017

Water-Soluble Chlorophyll Protein in Brassicaceae Plants is a Stress-Induced Chlorophyll-Binding Protein
journal, September 2001

  • Satoh, Hiroyuki; Uchida, Akira; Nakayama, Katsumi
  • Plant and Cell Physiology, Vol. 42, Issue 9
  • DOI: 10.1093/pcp/pce117

Editorial: Assembly of the Photosystem II Membrane-Protein Complex of Oxygenic Photosynthesis
journal, May 2017

Isolation and Characterization of Rhodobacter capsulatus Mutants Affected in Cytochromecbb 3 Oxidase Activity
journal, January 1998

Biosynthesis of Chlorophyll a in a Purple Bacterial Phototroph and Assembly into a Plant Chlorophyll–Protein Complex
journal, May 2016

  • Hitchcock, Andrew; Jackson, Philip J.; Chidgey, Jack W.
  • ACS Synthetic Biology, Vol. 5, Issue 9
  • DOI: 10.1021/acssynbio.6b00069

Water-Soluble Chlorophyll Protein in Brassicaceae Plants is a Stress-Induced Chlorophyll-Binding Protein
journal, September 2001

  • Satoh, Hiroyuki; Uchida, Akira; Nakayama, Katsumi
  • Plant and Cell Physiology, Vol. 42, Issue 9
  • DOI: 10.1093/pcp/pce117

Attachment of Noncognate Chromophores to CpcA of Synechocystis sp. PCC 6803 and Synechococcus sp. PCC 7002 by Heterologous Expression in Escherichia coli
journal, June 2011

  • Alvey, Richard M.; Biswas, Avijit; Schluchter, Wendy M.
  • Biochemistry, Vol. 50, Issue 22
  • DOI: 10.1021/bi200307s

Photosystem II Assembly: From Cyanobacteria to Plants
journal, April 2013

Bacterial Reaction Centers Purified with Styrene Maleic Acid Copolymer Retain Native Membrane Functional Properties and Display Enhanced Stability
journal, September 2014

  • Swainsbury, David J. K.; Scheidelaar, Stefan; van Grondelle, Rienk
  • Angewandte Chemie, Vol. 126, Issue 44
  • DOI: 10.1002/ange.201406412

Diverse Bacterial Genes Modulate Plant Root Association by Beneficial Bacteria
journal, December 2020

  • do Amaral, Fernanda Plucani; Tuleski, Thalita Regina; Pankievicz, Vania Carla Silva
  • mBio, Vol. 11, Issue 6
  • DOI: 10.1128/mbio.03078-20

Anaerobic chlorophyll isocyclic ring formation in Rhodobacter capsulatus requires a cobalamin cofactor
journal, June 2000

  • Gough, S. P.; Petersen, B. O.; Duus, J. O.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 12
  • DOI: 10.1073/pnas.97.12.6908

Identification and Characterization of a Novel Vitamin B 12 (Cobalamin) Biosynthetic Enzyme (CobZ) from Rhodobacter capsulatus , Containing Flavin, Heme, and Fe-S Cofactors
journal, November 2004

  • McGoldrick, Helen M.; Roessner, Charles A.; Raux, Evelyne
  • Journal of Biological Chemistry, Vol. 280, Issue 2
  • DOI: 10.1074/jbc.m411884200

Absence of the cbb 3 Terminal Oxidase Reveals an Active Oxygen-Dependent Cyclase Involved in Bacteriochlorophyll Biosynthesis in Rhodobacter sphaeroides
journal, May 2016

  • Chen, Guangyu E.; Canniffe, Daniel P.; Martin, Elizabeth C.
  • Journal of Bacteriology, Vol. 198, Issue 15
  • DOI: 10.1128/JB.00121-16

Regulation and function of tetrapyrrole biosynthesis in plants and algae
journal, September 2015

  • Brzezowski, Pawel; Richter, Andreas S.; Grimm, Bernhard
  • Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1847, Issue 9
  • DOI: 10.1016/j.bbabio.2015.05.007

Molecular mechanism of photosystem I assembly in oxygenic organisms
journal, September 2015

  • Yang, Huixia; Liu, Jun; Wen, Xiaogang
  • Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1847, Issue 9
  • DOI: 10.1016/j.bbabio.2014.12.011

Map of genes for carotenoid and bacteriochlorophyll biosynthesis in Rhodopseudomonas capsulata.
journal, January 1976

    Sort by:
    [ × clear filter / sort ]

    Works referencing / citing this record:

    Thawing out frozen metabolic accidents
    journal, January 2019

      Sort by:
      [ × clear filter / sort ]

      Figures / Tables found in this record:

      Fig. 1. (p. 2)figure
      Fig. 2. (p. 3)figure
      Fig. 3. (p. 4)figure
      Fig. 4. (p. 5)figure
        [ × clear filter / sort ]
        Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.

        Similar Records in DOE PAGES and OSTI.GOV collections: