Skip to main content

Advertisement

SpringerLink
Log in

In vitro activity of reconstituted rubisco enzyme from Gloeobacter violaceus

  • Published:
Journal of Biosciences Aims and scope Submit manuscript

Abstract

RuBisCO (Ribulose 1,5 bisphosphate carboxylase/oxygenase) by virtue of its dual specificity towards oxygen and carbon dioxide is an important rate-limiting step in photosynthesis and is believed to be the key factor for limited productivity of higher plants and algae. The photoautotrophic growth rate of cyanobacteria is a culmination of several factors including, rates of photosynthetic reactions, stress combating mechanisms and basic biomass generation metabolism in combination with optimal nutrient availability, irradiance, gaseous environment, etc. In case of cyanobacteria, the effect of RuBisCO in affecting the multiplication rate has been observed to show varied response. The current paper presents the RuBisCO activity of an early diverging cyanobacterium, Gloeobacter violaceus PCC 7421 and also compares the growth rates and RuBisCO activity of various cyanobacteria. A spectrophotometric estimation in a coupled enzyme assay system of the heterologous expressed G. violaceus PCC 7421 RuBisCO in E. coli, upon purification, revealed a carboxylation activity of LSu to be 5 nMol of phosphoglycerate min−1 mg−1 of protein, which is in coherence with the organism’s slow growth rate. Further, the in vitro complementation of RbcL with RbcS in presence of RbcX of G. violaceus facilitated partial reconstitution of the protein and was hence found to cause a four-fold enhancement in its specific activity. The unique characteristics of the primitive cyanobacteria, such as, absence of thylakoids, lack of several photosystem constituting genes, slow carboxylation rate, pose limitation for its rapid multiplication. The RuBisCO carboxylation rate is observed as not the sole but an important parameter for obtaining optimal cell multiplication rates in photo-autotrophically multiplying cyanobacteria.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2

Similar content being viewed by others

References

  • Anandan S and Golden SS 1997 cis-Acting sequences required for light-responsive expression of the psbDII gene in Synechococcus sp. strain PCC 7942. J. Bacteriol. 179 6865–6870

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arnon DI, Whatley FR and Allen MB 1954 Photosynthesis by isolated chloroplasts. II. Photosynthetic phosphorylation, the conversion of light into phosphate bond energy. J. Am. Chem. Soc. 76 6324–6329

    Article  CAS  Google Scholar 

  • Asato Y 2003 Toward an understanding of cell growth and the cell division cycle of unicellular photoautotrophic cyanobacteria. Cell. Mol. Life Sci. 60 663–687

    Article  CAS  PubMed  Google Scholar 

  • Bailey S, Melis A, Mackey KRM, Cardol P, Finazzi G, van Dijken G, Berg GM, Arrigo K, Shrager J and Grossman A 2008 Alternative photosynthetic electron flow to oxygen in marine Synechococcus. Biochim. Biophys. Acta 1777 269–276

    Article  CAS  PubMed  Google Scholar 

  • Bassham JA and Krause GH 1969 Free energy changes and metabolic regulation in steady-state photosynthetic carbon reduction. Biochim. Biophys. Acta Bioenergetics 189 207–221

    Article  CAS  Google Scholar 

  • Bernát G, Schreiber U, Sendtko E, Stadnichuk IN, Rexroth S, Rögner M and Koenig F 2012 Unique properties vs. common themes: the atypical cyanobacterium Gloeobacter violaceus PCC 7421 is capable of state transitions and blue-light-induced fluorescence quenching. Plant Cell Physiol. 53 528–542

    Article  PubMed  CAS  Google Scholar 

  • Bernstein HC, McClure RS, Hill EA, et al. 2016 Unlocking the Constraints of Cyanobacterial Productivity: Acclimations Enabling Ultrafast Growth. mBio 7 e00949-16

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Binder BJ and Chisholm SW 1990 Relationship between DNA cycle and growth rate in Synechococcus sp. strain PCC 6301. J. Bacteriol. 172 2313–2319

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bracher A, Hauser T, Liu C, Hartl FU and Hayer-Hartl M 2015 Structural analysis of the rubisco-assembly chaperone RbcX-II from Chlamydomonas reinhardtii. PLoS ONE

    Article  PubMed  PubMed Central  Google Scholar 

  • Bryant DA, Cohen-Bazire G and Glazer AN 1981 Characterization of the biliproteins of Gloeobacter violaceus chromophore content of a cyanobacterial phycoerythrin carrying phycourobilin chromophore. Arch. Microbiol. 129 190–198

    Article  CAS  Google Scholar 

  • Burnap RL 2015 Systems and photosystems: cellular limits of autotrophic productivity in cyanobacteria. Front. Bioeng. Biotechnol. 3 1

    Article  PubMed  PubMed Central  Google Scholar 

  • Calvin M and Benson AA 1948 The path of carbon in photosynthesis. Science 107 476–480

    Article  CAS  PubMed  Google Scholar 

  • Cloney LP, Bekkaoui DR and Hemmingsen SM 1993 Co-expression of plastid chaperonin genes and a synthetic plant Rubisco operon in Escherichia coli. Plant Mol. Biol. 23 1285–1290

    Article  CAS  PubMed  Google Scholar 

  • Colangeli R, Heijbel A, Williams AM, Manca C, Chan J, Lyashchenko K and Laura Gennaro M 1998 Three-step purification of lipopolysaccharide-free, polyhistidine-tagged recombinant antigens of Mycobacterium tuberculosis. J. Chromatogr. B: Biomed. Sci. App. 714 223–235

    Article  CAS  Google Scholar 

  • Daniell H, Torres-Ruiz JA, Inamdar A and McFadden BA 1988 Amplified expression of ribulose bisphosphate carboxylase/oxygenase in pBR322-transformants of Anacystis nidulans. Arch. Microbiol. 151 59–64

    Article  Google Scholar 

  • De Bernardez Clark E 1998 Refolding of recombinant proteins. Curr. Opin. Biotechnol. 9 157–163

    Article  CAS  Google Scholar 

  • Dietz K-J and Heber U 1984 Rate-limiting factors in leaf photosynthesis. I. Carbon fluxes in the calvin cycle. Biochim. Biophys. Acta Bioenergetics 767 432–443

    Article  CAS  Google Scholar 

  • Du Y-C, Nose A, Kawamitsu Y, Murayama S, Wasano K and Uchida Y 1996 An improved spectrophotometric determination of the activity of ribulose 1, 5-bishosphate carboxylase. Jap J. Crop Sci. 65 714–721

    Article  CAS  Google Scholar 

  • Farquhar GD, von Caemmerer S and Berry JA 1980 A biochemical model of photosynthetic CO2 assimilation in leaves of C3 species. Planta 149 78–90

    Article  CAS  PubMed  Google Scholar 

  • Furbank RT, Chitty JA, Von Caemmerer S and Jenkins CLD 1996 Antisense RNA inhibition of rbcs gene expression reduces rubisco level and photosynthesis in the C4 Plant Flaveria bidentis. Plant Physiol. 111 725–734

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Greene DN, Whitney SM and Matsumura I 2007 Artificially evolved Synechococcus PCC6301 Rubisco variants exhibit improvements in folding and catalytic efficiency. Biochem. J. 404 517–524

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gubernator B, Bartoszewski R, Kroliczewski J, Wildner G and Szczepaniak A 2008 Ribulose-1,5-bisphosphate carboxylase/oxygenase from thermophilic cyanobacterium Thermosynechococcus elongatus. Photosynthesis Res. 95 101–109

    Article  CAS  Google Scholar 

  • Guglielmi G, Cohen-Bazire G and Bryant DA 1981 The structure of Gloeobacter violaceus and its phycobilisomes. Arch. Microbiol. 129 181–189

    Article  CAS  Google Scholar 

  • Gupta RS and Mathews DW 2010 Signature proteins for the major clades of Cyanobacteria. BMC Evol. Biol. 10 24

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Gurevitz M, Somerville CR and McIntosh L 1985 Pathway of assembly of ribulose bisphosphate carboxylase/oxygenase from Anabaena 7120 expressed in Escherichia coli. Proc. Nat. Acad. Sci. USA 82 6546–6550

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hagemann M 2011 Molecular biology of cyanobacterial salt acclimation. FEMS Microbiol. Rev. 35 87–123

    Article  CAS  PubMed  Google Scholar 

  • Herdman M, Faulkner BM and Carr NG 1970 Synchronous growth and genome replication in the blue-green alga Anacystis nidulans. Archiv Mikrobiologie 73 238–249

    Article  CAS  Google Scholar 

  • Hoffmann L, Komárek J and Kaštovský J 2005 System of Cyanoprokaryotes (Cyanobacteria) - State in 2004. Algological Studies 117 95–115

    Google Scholar 

  • Ihlenfeldt MJ and Gibson J 1975 CO2 fixation and its regulation in Anacystis nidulans (Synechococcus). Arch. Microbiol. 102 13–21

    Article  CAS  PubMed  Google Scholar 

  • Imamura S and Asayama M 2009 Sigma factors for cyanobacterial transcription. Gene Regul. Syst. Biol. 3 65–87

    CAS  Google Scholar 

  • Iwaki T, Haranoh K, Inoue N, Kojima K, Satoh R, Nishino T, Wada S, Ihara H, Tsuyama S, Kobayashi H and Wadano A 2006 Expression of foreign type I ribulose-1,5-bisphosphate carboxylase/ oxygenase (EC 4.1.1.39) stimulates photosynthesis in cyanobacterium Synechococcus PCC7942 cells. Photosynthesis Res. 88 287–297

    Article  CAS  Google Scholar 

  • Koay TW, Wong HL and Lim BH 2016 Engineering of chimeric eukaryotic/bacterial Rubisco large subunits in Escherichia coli. Genes Genet. Syst. 91 139–150

    Article  CAS  PubMed  Google Scholar 

  • Kreel NE and Tabita FR 2007 Substitutions at methionine 295 of Archaeoglobus fulgidus ribulose-1,5-bisphosphate carboxylase/oxygenase affect oxygen binding and CO2/O2 specificity. J. Biol. Chem. 282 1341–1351

    Article  CAS  PubMed  Google Scholar 

  • Larimer FW and Soper TS 1993 Overproduction of Anabaena 7120 ribulose-bisphosphate carboxylase/oxygenase in Escherichia coli. Gene 126 85–92

    Article  CAS  PubMed  Google Scholar 

  • Lee B and Tabita FR 1990 Purification of recombinant ribulose-1,5-bisphosphate carboxylase/oxygenase large subunits suitable for reconstitution and assembly of active L8S8 enzyme. Biochemistry 29 9352–9357

    Article  CAS  PubMed  Google Scholar 

  • Lepp PW and Schmidt TM 1998 Nucleic acid content of Synechococcus spp. during growth in continuous light and light/dark cycles. Arch. Microbiol. 170 201–207

    Article  CAS  PubMed  Google Scholar 

  • Li LA and Tabita FR 1997 Maximum activity of recombinant ribulose 1,5-bisphosphate carboxylase/oxygenase of Anabaena sp. strain CA requires the product of the rbcX gene. J. Bacteriol. 179 3793–3796

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li R and Golden SS 1993 Enhancer activity of light-responsive regulatory elements in the untranslated leader regions of cyanobacterial psbA genes. Proc. Nat. Acad. Sci. USA 90 11678–11682

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liang F and Lindblad P 2017 Synechocystis PCC 6803 overexpressing RuBisCO grow faster with increased photosynthesis. Metab. Engin. Commun. 4 29–36

    Article  Google Scholar 

  • MacKenzie TDB and Campbell DA 2005 Cyanobacterial Acclimation to rapidly fluctuating light is constrained by inorganic carbon status1. J. Phycol. 41 801–811

    Article  CAS  Google Scholar 

  • Marcus Y, Altman-Gueta H, Finkler A and Gurevitz M 2005 Mutagenesis at two distinct phosphate-binding sites unravels their differential roles in regulation of rubisco activation and catalysis. J. Bacteriol. 187 4222–4228

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marcus Y, Altman-Gueta H, Wolff Y and Gurevitz M 2011 Rubisco mutagenesis provides new insight into limitations on photosynthesis and growth in Synechocystis PCC6803. J. Exp. Bot. 62 4173–4182

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Meeks JC, Wycoff KL, Chapman JS and Enderlin CS 1983 Regulation of expression of nitrate and dinitrogen assimilation by Anabaena Species. Appl. Environ. Microbiol. 45 1351–1359

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mori T, Binder B and Johnson CH 1996 Circadian gating of cell division in cyanobacteria growing with average doubling times of less than 24 hours. Proc. Nat. Acad. Sci. USA 93 10183–10188

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nakamura Y, Kaneko T, Sato S, et al. 2003 Complete genome structure of Gloeobacter violaceus PCC 7421; a cyanobacterium that lacks thylakoids. DNA Res. 10 137–145

    Article  CAS  PubMed  Google Scholar 

  • Nakano T, Ashida H, Mizohata E, Matsumura H and Yokota A 2010 An evolutionally conserved Lys122 is essential for function in Rhodospirillum rubrum bona fide RuBisCO and Bacillus subtilis RuBisCO-like protein. Biochem. Biophys. Res. Commun. 392 212–216

    Article  CAS  PubMed  Google Scholar 

  • Nogales J, Gudmundsson S, Knight EM, Palsson BO and Thiele I 2012 Detailing the optimality of photosynthesis in cyanobacteria through systems biology analysis. Proc. Nat. Acad. Sci. 109 2678–2683

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Parrott LM and Slater JH 1980 The DNA, RNA and protein composition of the cyanobacterium Anacystis nidulans grown in light- and carbon dioxide-limited chemostats. Arch. Microbiol. 127 53–58

    Article  CAS  PubMed  Google Scholar 

  • Prasanna R, Kumar R, Sood A, Prasanna BM and Singh PK 2006 Morphological, physiochemical and molecular characterization of Anabaena strains. Microbiol. Res. 161 187–202

    Article  CAS  PubMed  Google Scholar 

  • Raven JA 2011 The cost of photoinhibition. Physiologia Plantarum 142 87–104

    Article  CAS  PubMed  Google Scholar 

  • Raven JA and Larkum AWD 2007 Are there ecological implications for the proposed energetic restrictions on photosynthetic oxygen evolution at high oxygen concentrations? Photosynthesis Res. 94 31–42

    Article  CAS  Google Scholar 

  • Rippka R, Waterbury J and Cohen-Bazire G 1974 A cyanobacterium which lacks thylakoids. Arch. Microbiol. 100 419–436

    Article  CAS  Google Scholar 

  • Selstam E and Campbell D 1996 Membrane lipid composition of the unusual cyanobacterium Gloeobacter violaceus sp. PCC 7421; which lacks sulfoquinovosyl diacylglycerol. Arch. Microbiol. 166 132–135

    Article  CAS  Google Scholar 

  • Seo P-S and Yokota A 2003 The phylogenetic relationships of cyanobacteria inferred from 16S rRNA, gyrB, rpoC1 and rpoD1 gene sequences. J. Gen. Appl. Microbiol. 49 191–203

    Article  CAS  PubMed  Google Scholar 

  • Smith SA and Tabita FR 2004 Glycine 176 Affects catalytic properties and stability of the Synechococcus sp. Strain PCC6301 Ribulose-1,5-bisphosphate carboxylase/oxygenase. J. Biol. Chem. 279 25632–25637

    Article  CAS  PubMed  Google Scholar 

  • Snir A, Gurevitz M and Marcus Y 2006 Alterations in Rubisco activity and in stomatal behavior induce a daily rhythm in photosynthesis of aerial leaves in the amphibious-plant Nuphar lutea. Photosynthesis Res. 90 233–242

    Article  CAS  Google Scholar 

  • Stitt M, Quick WP, Schurr U, Schulze ED, Rodermel SR and Bogorad L 1991 Decreased ribulose-1,5-bisphosphate carboxylase-oxygenase in transgenic tobacco transformed with ‘antisense’ rbcS: II. Flux-control coefficients for photosynthesis in varying light, CO2, and air humidity. Planta 183 555–566

    Article  CAS  PubMed  Google Scholar 

  • Tcherkez GGB, Farquhar GD and Andrews TJ 2006 Despite slow catalysis and confused substrate specificity, all ribulose bisphosphate carboxylases may be nearly perfectly optimized. Proc. Nat. Acad. Sci. USA 103 7246–7251

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vermass WFJ, Rutherford AW and Hansson Ö 1988 Site-directed mutagenesis in photosystem II of the cyanobacterium Synechocystis sp. PCC 6803 Donor D is a tyrosine residue in the D2 protein. Proc. Nat. Acad. Sci. USA 85 8477–8481

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Whitney SM and Sharwood RE 2007 Linked Rubisco subunits can assemble into functional oligomers without impeding catalytic performance. J. Biol. Chem. 282 3809–3818

    Article  CAS  PubMed  Google Scholar 

  • Whitney SM, Baldet P, Hudson GS and Andrews TJ 2001 Form I Rubiscos from non-green algae are expressed abundantly but not assembled in tobacco chloroplasts. Plant J. 26 535–547

    Article  CAS  PubMed  Google Scholar 

  • Yamaoka T, Satoh K and Katoh S 1978 Photosynthetic activities of a thermophilic blue-green alga. Plant Cell Physiol. 19 943–954

    Article  CAS  Google Scholar 

  • Yu J, Liberton M, Cliften PF, Head RD, Jacobs JM, Smith RD, Koppenaal DW, Brand JJ and Pakrasi HB 2015 Synechococcus elongatus UTEX 2973; a fast growing cyanobacterial chassis for biosynthesis using light and CO2. Sci. Rep. 5 8132

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zavřel T, Sinetova MA, Búzová D, Literáková P and Červený J 2015 Characterization of a model cyanobacterium Synechocystis sp. PCC 6803 autotrophic growth in a flat-panel photobioreactor. Eng. Life Sci. 15 122–132

    Article  CAS  Google Scholar 

  • Zilliges Y and Dau H 2016 Unexpected capacity for organic carbon assimilation by Thermosynechococcus elongatus, a crucial photosynthetic model organism. FEBS Lett. 590 962–970

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors are grateful to the Birla Institute of Technology and Science, Pilani, Rajasthan, India, for providing infrastructural and logistic support. GKS is thankful to the UGC-BSR for her fellowship. PN is thankful to CSIR for Senior Research fellowship. VT is thankful to DST-inspire fellowship program of DST, India.

Funding

This work was supported by SERB fast track project SERC/LS-0141/2010 sanctioned by the Government of India to SM.

Author information

Authors and Affiliations

  1. Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad, Telangana, India

    Gurpreet Kaur Sidhu

  2. Department of Biological Sciences, Birla Institute of Technology and Science, Pilani Campus, Pilani, Rajasthan, India

    Gurpreet Kaur Sidhu, Panchsheela Nogia, Vandana Tomar, Rajesh Mehrotra & Sandhya Mehrotra

  3. Department of Biotechnology, Guru Nanak Dev University, Amritsar, Punjab, India

    Panchsheela Nogia

  4. Department of Biological Sciences, Birla Institute of Technology and Science, Goa Campus, Pilani, Goa, India

    Rajesh Mehrotra & Sandhya Mehrotra

Authors
  1. Gurpreet Kaur Sidhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Panchsheela Nogia
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vandana Tomar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rajesh Mehrotra
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sandhya Mehrotra
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sandhya Mehrotra.

Additional information

Communicated by B J Rao.

Corresponding editor: B J Rao

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sidhu, G.K., Nogia, P., Tomar, V. et al. In vitro activity of reconstituted rubisco enzyme from Gloeobacter violaceus. J Biosci 46, 67 (2021). https://doi.org/10.1007/s12038-021-00188-w

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s12038-021-00188-w

Keywords

Search

Navigation