Biosynthesis of Cyanobacterial Phycobiliproteins in Escherichia Coli: Chromophorylation Efficiency and Specificity of All Bilin Lyases from Synechococcus Sp. Strain PCC 7002

Overview

Journal Appl Environ Microbiol

Specialties Environmental Health
Microbiology

Date 2010 Mar 16

PMID 20228104

Citations 32

Authors

Avijit Biswas

Yasmin M Vasquez

Tierna M Dragomani

Monica L Kronfel

Shervonda R Williams

Richard M Alvey

Donald A Bryant

Wendy M Schluchter

Affiliations

Soon will be listed here.

Abstract

Phycobiliproteins are water-soluble, light-harvesting proteins that are highly fluorescent due to linear tetrapyrrole chromophores, which makes them valuable as probes. Enzymes called bilin lyases usually attach these bilin chromophores to specific cysteine residues within the alpha and beta subunits via thioether linkages. A multiplasmid coexpression system was used to recreate the biosynthetic pathway for phycobiliproteins from the cyanobacterium Synechococcus sp. strain PCC 7002 in Escherichia coli. This system efficiently produced chromophorylated allophycocyanin (ApcA/ApcB) and alpha-phycocyanin with holoprotein yields ranging from 3 to 12 mg liter(-1) of culture. This heterologous expression system was used to demonstrate that the CpcS-I and CpcU proteins are both required to attach phycocyanobilin (PCB) to allophycocyanin subunits ApcD (alpha(AP-B)) and ApcF (beta(18)). The N-terminal, allophycocyanin-like domain of ApcE (L(CM)(99)) was produced in soluble form and was shown to have intrinsic bilin lyase activity. Lastly, this in vivo system was used to evaluate the efficiency of the bilin lyases for production of beta-phycocyanin.

Citing Articles

Functional Modification of Cyanobacterial Phycobiliprotein and Phycobilisomes through Bilin Metabolism Control.

Sato M, Kawaguchi T, Maeda K, Watanabe M, Ikeuchi M, Narikawa R ACS Synth Biol. 2024; 13(8):2391-2401.

PMID: 39038807 PMC: 11334911. DOI: 10.1021/acssynbio.4c00094.

Recent Progress of Natural and Recombinant Phycobiliproteins as Fluorescent Probes.

Chen H, Deng J, Li L, Liu Z, Sun S, Xiong P Mar Drugs. 2023; 21(11).

PMID: 37999396 PMC: 10672124. DOI: 10.3390/md21110572.

Loss of Biliverdin Reductase Increases Oxidative Stress in the Cyanobacterium sp. PCC 7002.

Schluchter W, Babin C, Liu X, Bieller A, Shen G, Alvey R Microorganisms. 2023; 11(10).

PMID: 37894251 PMC: 10608806. DOI: 10.3390/microorganisms11102593.

The Unique Light-Harvesting System of the Algal Phycobilisome: Structure, Assembly Components, and Functions.

Li X, Hou W, Lei J, Chen H, Wang Q Int J Mol Sci. 2023; 24(11).

PMID: 37298688 PMC: 10254012. DOI: 10.3390/ijms24119733.

Elucidating the origins of phycocyanobilin biosynthesis and phycobiliproteins.

Rockwell N, Martin S, Lagarias J Proc Natl Acad Sci U S A. 2023; 120(17):e2300770120.

PMID: 37071675 PMC: 10151467. DOI: 10.1073/pnas.2300770120.

References

Glazer A . Light guides. Directional energy transfer in a photosynthetic antenna. J Biol Chem. 1989; 264(1):1-4. View

Yang Y, Ge B, Guan X, Zhang W, Qin S . Combinational biosynthesis of a fluorescent cyanobacterial holo-alpha-allophycocyanin in Escherichia coli. Biotechnol Lett. 2008; 30(6):1001-4. DOI: 10.1007/s10529-008-9644-2. View

Zhao K, Zhang J, Tu J, Bohm S, Ploscher M, Eichacker L . Lyase activities of CpcS- and CpcT-like proteins from Nostoc PCC7120 and sequential reconstitution of binding sites of phycoerythrocyanin and phycocyanin beta-subunits. J Biol Chem. 2007; 282(47):34093-103. DOI: 10.1074/jbc.M703038200. View

Ge B, Sun H, Feng Y, Yang J, Qin S . Functional biosynthesis of an allophycocyan beta subunit in Escherichia coli. J Biosci Bioeng. 2009; 107(3):246-9. DOI: 10.1016/j.jbiosc.2008.11.014. View

Wedemayer G, Kidd D, Glazer A . Cryptomonad biliproteins: Bilin types and locations. Photosynth Res. 2013; 48(1-2):163-70. DOI: 10.1007/BF00041006. View

Fu E, Friedman L, Siegelman H . Mass-spectral identification and purification of phycoerythrobilin and phycocyanobilin. Biochem J. 1979; 179(1):1-6. PMC: 1186587. DOI: 10.1042/bj1790001. View

Tooley A, Cai Y, Glazer A . Biosynthesis of a fluorescent cyanobacterial C-phycocyanin holo-alpha subunit in a heterologous host. Proc Natl Acad Sci U S A. 2001; 98(19):10560-5. PMC: 58505. DOI: 10.1073/pnas.181340998. View

Tooley A, Glazer A . Biosynthesis of the cyanobacterial light-harvesting polypeptide phycoerythrocyanin holo-alpha subunit in a heterologous host. J Bacteriol. 2002; 184(17):4666-71. PMC: 135287. DOI: 10.1128/JB.184.17.4666-4671.2002. View

Fairchild C, Glazer A . Nonenzymatic bilin addition to the alpha subunit of an apophycoerythrin. J Biol Chem. 1994; 269(46):28988-96. View

10.

Glazer A . Phycobiliproteins. Methods Enzymol. 1988; 167:291-303. DOI: 10.1016/0076-6879(88)67034-0. View

11.

Zhang W, Guan X, Yang Y, Ge B, Chen H, Li F . Biosynthesis of fluorescent allophycocyanin alpha-subunits by autocatalysis in Escherichia coli. Biotechnol Appl Biochem. 2008; 52(Pt 2):135-40. DOI: 10.1042/BA20070187. View

12.

Capuano V, Braux A, de Marsac N, Houmard J . The "anchor polypeptide" of cyanobacterial phycobilisomes. Molecular characterization of the Synechococcus sp. PCC 6301 apce gene. J Biol Chem. 1991; 266(11):7239-47. View

13.

Zhao K, Su P, Bohm S, Song B, Zhou M, Bubenzer C . Reconstitution of phycobilisome core-membrane linker, LCM, by autocatalytic chromophore binding to ApcE. Biochim Biophys Acta. 2004; 1706(1-2):81-7. DOI: 10.1016/j.bbabio.2004.09.008. View

14.

Zhou J, Gasparich G, Stirewalt V, de Lorimier R, Bryant D . The cpcE and cpcF genes of Synechococcus sp. PCC 7002. Construction and phenotypic characterization of interposon mutants. J Biol Chem. 1992; 267(23):16138-45. View

15.

Cornejo J, Willows R, Beale S . Phytobilin biosynthesis: cloning and expression of a gene encoding soluble ferredoxin-dependent heme oxygenase from Synechocystis sp. PCC 6803. Plant J. 1998; 15(1):99-107. DOI: 10.1046/j.1365-313x.1998.00186.x. View

16.

Tu J, Kupka M, Bohm S, Ploscher M, Eichacker L, Zhao K . Intermediate binding of phycocyanobilin to the lyase, CpeS1, and transfer to apoprotein. Photosynth Res. 2007; 95(2-3):163-8. DOI: 10.1007/s11120-007-9251-4. View

17.

Brejc K, Ficner R, Huber R, Steinbacher S . Isolation, crystallization, crystal structure analysis and refinement of allophycocyanin from the cyanobacterium Spirulina platensis at 2.3 A resolution. J Mol Biol. 1995; 249(2):424-40. DOI: 10.1006/jmbi.1995.0307. View

18.

Shen G, Saunee N, Williams S, Gallo E, Schluchter W, Bryant D . Identification and characterization of a new class of bilin lyase: the cpcT gene encodes a bilin lyase responsible for attachment of phycocyanobilin to Cys-153 on the beta-subunit of phycocyanin in Synechococcus sp. PCC 7002. J Biol Chem. 2006; 281(26):17768-78. DOI: 10.1074/jbc.M602563200. View

19.

Montgomery B, Lagarias J . Phytochrome ancestry: sensors of bilins and light. Trends Plant Sci. 2002; 7(8):357-66. DOI: 10.1016/s1360-1385(02)02304-x. View

20.

Frankenberg N, Lagarias J . Phycocyanobilin:ferredoxin oxidoreductase of Anabaena sp. PCC 7120. Biochemical and spectroscopic. J Biol Chem. 2003; 278(11):9219-26. DOI: 10.1074/jbc.M211643200. View