Abstract
Arthrospira platensis is an economically important cyanobacterium; and it has been used widely in food and pharmaceutical industries. The phycocyanin (PC) from A. platensis is extremely valuable in medicine and molecular biology due to its antioxidation and anti-tumoring activity and applicability as fluorescence protein tag. In present study, two recombinant plasmids, one contained the phycocyanobilin (PCB)-producing genes (hox1 and pcyA) while the other contained the phycobiliprotein gene (cpcB) and the lyase gene (either cpcS/U or cpcT), were constructed and synchronically transferred into E. coli in order to test the the activities of relevant lyases for catalysing PCB addition to CpcB during synthesizing fluorescent PC holo-β-subunit (β-PC) of A. platensis. As was evidenced by the fluorescence emitted at a peak specific for PC, CpcB was successfully synthesized in E. coli, to which co-expressed PCBs attached though at a relatively low efficiency. The results showed that the attachment of PCBs to CpcB were carried out mainly by co-expressed CpcS/U but CpcB also showed some autocatalytic activity. Currently, no CpcT activity was detected in this E. coli expression system. Further studies will be conducted to improve the efficiency of fluorescent PC synthesis in E. coli.
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Arciero, D. M., Dallas, J. L., and Glazer, A. N., 1988. In vitro attachment of bilins to apophycocyanin. II. Determination of the structures of tryptic bilin peptides derived from the phycocyanobilin adduct. Journal of Biological Chemistry, 263: 18350–18357.
Benedetti, S., Benvenuti, F., Scoglio, S., and Canestrari, F., 2010. Oxygen radical absorbance capacity of phycocyanin and phycocyanobilin from the food supplement Aphanizomenon flos-aquae. Journal of Medicinal Food, 13(1): 223–227.
Biswas, A., Vasquez, Y. M., Dragomani, T. M., Kronfel, M. L., Williams, S. R., Alvey, R. M., Bryant, D. A., and Schluchter, W. M., 2010. Biosynthesis of cyanobacterial phycobiliproteins in Escherichia coli: Chromophorylation efficiency and specificity of all bilin lyases from Synechococcus sp. Strain PCC 7002. Applied and Environmental Microbiology, 76(9): 2729–2739.
Cobley, J. G., Clark, A. C., Weerasurya, S., Queseda, F. A., Xiao, J. Y., Bandrapali, N., D’Silva, I., Thounaojam, M., Oda, J. F., Sumiyoshi, T., and Chu, M. H., 2002. CpeR is an activator required for expression of the phycoerythrin operon(cpeBA) in the cyanobacterium Fremyella diplosiphon and is encoded in the phycoerythrin linker-polypeptide operon (cpeCDESTR). Molecular Microbiology, 44(6): 1517–1531.
Debreczeny, M. P., Sauer, K., Zhou, J., and Bryant, D. A., 1995. Comparison of calculated and experimentally resolved rate constants for excitation energy transfer in C-phycocyanin. 1. Monomers. Journal of Physical Chemistry, 99: 8412–8419.
Fairchild, C. D., Zhao, J. D., Zhou, J. H., Colson, S. E., Bryant, D. A., and Glazer, A. N., 1992. Phycocyanin α-subunit phycocyanobilin lyase. Proceedings of the National Academy of Sciences of the United States of America, 89: 7017–7021.
Fujisawa, T., Narikawa, R., Okamoto, S., Ehira, S., Yoshimura, H., Suzuki, I., Masuda, T., Mochimaru, M., Takaichi, S., Awai, K., Sekine, M., Horikawa, H., Yashiro, I., Omata, S., Takarada, H., Katano, Y., Kosugi, H., Tanikawa, S., Ohmori, K., Sato, N., Ikeuchi, M., Fujita, N., and Ohmori, M., 2010. Genomic structure of an economically important cyanobacterium, Arthrospira (Spirulina) platensis NIES-39. DNA Research, 17: 85–103
Ge, B. S., Sun, H. X., Feng, Y., Yang, J. Y., and Qin, S., 2009. Functional biosynthesis of an allophycocyan β-subunit in Escherichia coli. Journal of Bioscience and Bioengineering, 107(3): 246–249.
Glazer, A. N., 1985. Light harvesting by phycobilisomes. Annual Review of Biophysics and Biophysical Chemistry, 14: 47–77.
Guan, X. Y., Qin, S., Su, Z. L., Zhao, F. Q., Ge, B. S., Li, F. C., and Tang, X. X., 2007. Combinational biosynthesis of a fluorescent cyanobacterial holo-α-phycocyanin in Escherichia coli using one expression vector. Applied Biochemistry and Biotechnology, 142: 52–59.
Jung, L. J., Chan, C. F., and Glazer, A. N., 1995. Candidate genes for the phycoerythrocyanin α subunit lyase: Biochemical analysis of pecE and pecF tnterposon mutants. Journal of Biological Chemistry, 270: 12877–12884.
Landgraf, F. T., Forreite, C., Hurtado, P., Lamparter, T., and Hughes, J., 2001. Recombinant holophytochrome in Escherichia coli. FEBS Letters, 508: 459–462.
Li, H., and Sherman, L. A., 2002. Characterization of Synechocystis sp. Strain PCC 6803 and Δnbl mutants under nitrogen-deficient conditions. Archives of Microbiology, 178: 256–266.
MacColl, R., Csatorday, K., Berns, D. S., and Traeger, E., 1980. Chromophore interactions in allophycocyanin. Biochemistry-US, 19: 2817–2820.
Niu, J. F., Wang, G. C., Lin, X. Z., and Zhou, B. C., 2007. Large-scale recovery of C-phycocyanin from Spirulina platensis using expanded bed adsorption chromatography. Journal of Chromatography B, 850: 267–276.
Nyman, E. S., and Hynninen, P. H., 2004. Research advances in the use of tetrapyrrolic photosensitizers for photodynamic therapy. Photochemistry and Photobiology, 73: 1–2.
Ong, L. J., and Glazer, A. N., 1988. Structural studies of phycobiliproteins in unicellular marine cyanobacteria. In: Light-Energy Transduction in Photosynthesis: Higher Plant and Bacterial Models. Stevens, S. E., and Jr Bryant, D. A., eds., American Society of Plant Physiologists, Rockville, MD, 102–121.
Rodríguez-Sánchez, R., Ortiz-Butrón, R., Blas-Valdivia, V., Hernández-García, A., and Cano-Europa, E., 2012. Phycobiliproteins or C-phycocyanin of Arthrospira (Spirulina) maxima protect against HgCl2-caused oxidative stress and renal damage. Food Chemistry, 135(4): 2359–2365.
Saunée, N. A., Williams, S. R., Bryant, D. A., and Schluchter, W. M., 2008. Biogenesis of phycobiliproteins II. CpcS-I and CpcU comprise the heterodimeric bilin lyase that attaches phycocyanobilin to Cys-82 of β-phycocyanin and Cys-81 of allophycocyanin subunits in Synechococcus sp. PCC 7002. Journal of Biological Chemistry, 283(12): 7513–7522.
Scheer, H., and Zhao, K. H., 2008. Biliprotein maturation: the chromophore attachment. Molecular Microbiology, 68(2): 263–276.
Schmidt, M., Patel, A., Zhao, Y., and Reuter, W., 2007. Structural basis for the photochemistry of α-Phycoerythrocyanin. Biochemistry-US., 46: 416–423.
Shen, G. Z., Saunée, N. A., Gallo, E. F., Begovic, Z., Schluchter, W. M., and Bryant, D. A., 2004. Identification of novel phycobiliprotein lyases in cyanobacteria. In: Photosynthesis 2004 Light-harvesting Systems Workshop. Niederman, R. A., et al., eds., Saint Adele, Québec, 14–15.
Shen, G. Z., Saunée, N. A., Williams, S. R., Gallo, E. F., Schluchter, W. M., and Bryant, D. A., 2006. 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 β-Subunit of phycocyanin in Synechococcus sp. PCC 7002. Journal of Biological Chemistry, 281(26): 17768–17778.
Shen, G. Z., Schluchter, W. M., and Bryant, D. A., 2008. Biogenesis of phycobiliproteins I. cpcS-I and cpcU mutants of the cyanobacterium Synechococcus sp. PCC 7002 define a heterodimeric phycocyanobilin lyase specific for β-phycocyanin and allophycocyanin subunits. Journal of Biological Chemistry, 283(12): 7503–7512.
Sidler, A., 1994. Phycobilisome and phycobiliprotein structures. In: The Molecular Biology of Cyanobacteria. Bryant, D. A., ed., Kluwer, Dordrecht, 139–216.
Storf, M., Parbel, A., Meyer, M., Strohmann, B., Scheer, H., Deng, M. G., Zheng, M., Zhou, M., and Zhao, K. H., 2001. Chromophore attachment to biliproteins: Specificity of PecE/PecF, a lyase-Isomerase for the photoactive 31-Cys-α84-phycoviolobilin chromophore of phycoerythrocyanin. Biochemistry-US, 40: 12444–12456.
Tooley, A. J., and Glazer, A. N., 2002. Biosynthesis of the cyanobacterial light-harvesting polypeptide phycoerythrocyanin holo-α-subunit in a heterologous host. Journal of Bacteriology, 184(17): 4666–4671.
Tooley, A. J., Cai, Y. A., and Glazer, A. N., 2001. Biosynthesis of a fluorescent cyanobacterial C-phycocyanin holo-α subunit in a heterologous host. Proceedings of the National Academy of Sciences of the United States of America, 98(19): 10560–10565.
Yi, J. J., Zang, X. N., Zhang, X. C., Yuan, D. Y., Zhao, B. R., and Tang, L., 2011. Recombinant expression of a fluorescent phycocyanin holo-α-subunit from Arthrospira platensis in Escherichia coli. Periodical of Ocean University of China, 41(5): 59–65 (in Chinese).
Zhao, K. H., Deng, M. G., Zheng, M., Zhou, M., Parbel, A., Storf, M., Meyer, M., Strohmann, B., and Scheer, H., 2000. Novel activity of a phycobiliprotein lyase: Both the attachment of phycocyanobilin and the isomerization to phycoviolobilin are catalyzed by the proteins PecE and PecF encoded by the phycoerythrocyanin operon. FEBS Letters, 469: 9–13.
Zhao, K. H., Su, P., Tu, J. M., Wang, X., Liu, H., Plöscher, M., Eichacker, L., Yang, B., Zhou, M., and Scheer, H., 2007a. Phycobilin: Cystein-84 biliprotein lyase, a near-universal lyase for cysteine-84-binding sites in cyanobacterial phycobiliproteins. Proceedings of the National Academy of Sciences of the United States of America, 104(36): 14300–14305.
Zhao, K. H., Wu, D., Wang, L., Zhou, M., Storf, M., Bubenzer, C., Strohmann, B., and Scheer, H., 2002. Characterization of phycoviolobilin phycoerythrocyanin-α84-cystein-lyase-(isomerizing) from Mastigocladus laminosus. European Journal of Biochemistry, 269: 4542–4550.
Zhao, K. H., Zhang, J., Tu, J. M., Böhm, S., Plöscher, M., Eichacker, L., Bubenzer, C., Scheer, H., Wang, X., and Zhou, M., 2007b. Lyase activities of CpcS- and CpcT-like proteins from Nostoc PCC7120 and sequential reconstitution of binding sites of phycoerythrocyanin and phycocyanin β-subunits. Journal of Biological Chemistry, 282(47): 34093–34103.
Zhao, K. H., Zhu, J. P., Song, B., Zhou, M., Storf, M., Böhm, S., Bubenzer, C., and Scheer, H., 2004. Nonenzymatic chromophore attachment in biliproteins: Conformational control by the detergent Triton X-100. Biochimica et Biophysica Acta (BBA)-Bioenergetics, 1657: 131–145.
Zhao, K.H., Su, P., Li, J., Tu, J.M., Zhou, M., Bubenzer, C., and Scheer, H., 2006. Chromophore attachment to phycobiliprotein β-subunits: Phycocyanobilin cysteine-β84 phycobiliprotein lyase activity of CpeS-like protein from Anabaena sp. PCC7120. Journal of Biological Chemistry, 281(13): 8573–8581.
Zhou, J., Gasparich, G. E., Stirewalt, V. L., de Lorimier, R., and Bryant, D. A., 1992. The cpcE and cpcF genes of Synechococcus sp. PCC 7002. Construction and phenotypic characterization of interposon mutants. Journal of Biological Chemistry, 267: 16138–16145.
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Yi, J., Xu, D., Zang, X. et al. Lyase activities of heterologous CpcS and CpcT for phycocyanin holo-β-subunit from Arthrospira platensis in Escherichia coli . J. Ocean Univ. China 13, 497–502 (2014). https://doi.org/10.1007/s11802-014-2161-0
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DOI: https://doi.org/10.1007/s11802-014-2161-0